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EXAM I Page 1 2012 ASTRO RADIATION/CANCER BIOLOGY PRACTICE EXAMINATION AND STUDY GUIDE Produced by the Radiation/Cancer Biology Practice Examination and Study Guide Subcommittee of the ASTRO Radiation and Cancer Biology Committee Please address all correspondence to: Dr. Barry S. Rosenstein Box 1236 Department of Radiation Oncology Mount Sinai School of Medicine One Gustave Levy Place New York, NY 10029 Tel: (212) 241-9408 Fax: (212) 996-8927 Email: barry.rosenstein@mssm.edu © 2011 American Society for Radiation Oncology. All rights reserved. Reproduction or dissemination of any portion of this publication is strictly prohibited unless express written authorization is first obtained from the American Society for Radiation Oncology, 8280 Willow Oaks Corporate Drive, Suite 500, Fairfax, VA 22031. Page 2 2 Editor-in-Chief, Chair of Subcommittee Barry S. Rosenstein, Ph.D. Mount Sinai School of Medicine NYU School of Medicine Members of Subcommittee Christopher Barker, M.D. Memorial Sloan Kettering Cancer Center Gayle E. Woloschak, Ph.D. Northwestern University Medical School Associate Editors Elizabeth Balcer-Kubiczek, Ph.D. University of Maryland School of Medicine Christopher Barker, M.D. Memorial Sloan Kettering Cancer Center Michael C. Joiner, Ph.D. Karmanos Cancer Institute, Wayne State University School of Medicine David G. Kirsch, M.D., Ph.D. Duke University Medical Center Brian Marples, Ph.D. William Beaumont Hospital Michael E.C. Robbins, Ph.D. Wake Forest University School of Medicine Juong G. Rhee Ph.D. University of Maryland School of Medicine Navesh K. Sharma, Ph.D., D.O. University of Maryland School of Medicine Zhiyuan Shen, M.D., Ph.D. UMDNJ-Robert Wood Johnson Medical School Michael D. Story, Ph.D. University of Texas Southwestern Medical Center Gayle E. Woloschak, Ph.D. Northwestern University Medical School Brad G. Wouters, Ph.D. Ontario Institute for Cancer Research Associate Editors (2006-2011) Kathryn D. Held, Ph.D. Harvard Medical School Massachusetts General Hospital Sara Rockwell, Ph.D. Yale University School of Medicine Jacqueline Williams, Ph.D. University of Rochester Medical Center Elaine M. Zeman, Ph.D. University of North Carolina School of Medicine Page 3 3 Contributors Joan Allalunis-Turner, Ph.D. Cross Cancer Institute Sally A. Amundson, Ph.D. Columbia University Elizabeth K. Balcer-Kubiczek, Ph.D. University of Maryland School of Medicine Kevin A. Camphausen, M.D. National Cancer Institute/NIH Theodore L. DeWeese, M.D. Johns Hopkins University School of Medicine Evan B. Douple, Ph.D. The National Academies Nuclear and Radiation Studies Board Adriana Haimovitz-Friedman, Ph.D. Memorial Sloan-Kettering Cancer Center Dennis E. Hallahan, M.D. Vanderbilt University Martin Hauer-Jensen, M.D., Ph.D. University of Arkansas for Medical Sciences Kathryn D. Held, Ph.D. Harvard Medical School Massachusetts General Hospital Richard P. Hill, Ph.D. Ontario Cancer Institute Rakesh K. Jain, Ph.D. Harvard Medical School Massachusetts General Hospital David Kirsch, M.D., Ph.D. Duke University School of Medicine Amy Kronenberg, Sc.D. Lawrence Berkeley National Laboratory William F. Morgan, Ph.D. University of Maryland John P. Murnane, Ph.D. University of California, San Francisco Peggy L. Olive, Ph.D. British Columbia Cancer Research Centre Martin Pruschy, Ph.D. University Hospital Zurich Sara Rockwell, Ph.D. Yale University School of Medicine Michael E.C. Robbins, Ph.D. Wake Forest University School of Medicine Joseph L. Roti Roti, Ph.D. Washington University School of Medicine Carolyn I. Sartor, M.D. University of North Carolina School of Medicine Joann Sweasy, Ph.D. Yale University School of Medicine Marie-Catherine Vozenin-Brotons Ph.D. Institut Gustave Roussy Gayle E. Woloschak, Ph.D. Northwestern University Medical School Henning Willers, M.D. Harvard Medical School Massachusetts General Hospital Jacqueline Williams, Ph.D. University of Rochester Medical Center Elaine M. Zeman, Ph.D. University of North Carolina School of Medicine Daniel Zips, Ph.D. Technical University Dresden Page 4 TABLE OF CONTENTS TOPIC PAGE NUMBER Table of Contents 4 Preface 6 Note on Protein and Gene Nomenclature 7 Questions I. Interaction of Radiation with Matter 9 II. Molecular Mechanisms of DNA Damage 11 III. Molecular Mechanisms of DNA Repair 13 IV. Chromosome and Chromatid Damage 17 V. Mechanisms of Cell Death 19 VI. Cell and Tissue Survival Assays 23 VII. Models of Cell Survival 24 VIII. Linear Energy Transfer 27 IX. Modifiers of Cell Survival: Oxygen Effect 28 X. Modifiers of Cell Survival: Repair 31 XI. Solid Tumor Assay Systems 33 XII. Tumor Microenvironment 35 XIII. Cell and Tissue Kinetics 40 XIV. Molecular Signaling 43 XV. Cancer 47 XVI. Total Body Irradiation 50 XVII. Clinically Relevant Normal Tissue Responses to Radiation 54 XVIII. Mechanisms of Normal Tissue Radiation Responses 58 XIX. Therapeutic Ratio 62 XX. Time, Dose, Fractionation 66 XXI. Brachytherapy 71 XXII. Radiobiological Aspects of Alternative Dose Delivery Systems 73 XXIII. Chemotherapeutic Agents and Radiation Therapy 75 XXIV. Radiosensitizers, Radioprotectors and Bioreductive Drugs 79 XXV. Hyperthermia 81 XXVI. Radiation Carcinogenesis 82 XXVII. Heritable Effects of Radiation 85 XXVIII. Radiation Effects in the Developing Embryo and Fetus 86 XXIX. Radiation Protection 87 XXX. Molecular Techniques Used in Radiation and Cancer Biology 90 XXXI. Molecular Imaging 92 Page 5 5 Answers, Explanations and References General References 95 I. Interaction of Radiation with Matter 98 II. Molecular Mechanisms of DNA Damage 100 III. Molecular Mechanisms of DNA Repair 102 IV. Chromosome and Chromatid Damage 107 V. Mechanisms of Cell Death 108 VI. Cell and Tissue Survival Assays 114 VII. Models of Cell Survival 115 VIII. Linear Energy Transfer 117 IX. Modifiers of Cell Survival: Oxygen Effect 118 X. Modifiers of Cell Survival: Repair 120 XI. Solid Tumor Assay Systems 122 XII. Tumor Microenvironment 123 XIII. Cell and Tissue Kinetics 128 XIV. Molecular Signaling 130 XV. Cancer 135 XVI. Total Body Irradiation 140 XVII. Clinically Relevant Normal Tissue Responses to Radiation 146 XVIII. Mechanisms of Normal Tissue Radiation Responses 152 XIX. Therapeutic Ratio 159 XX. Time, Dose, Fractionation 162 XXI. Brachytherapy 165 XXII. Radiobiological Aspects of Alternative Dose Delivery Systems167 XXIII. Chemotherapeutic Agents and Radiation Therapy 169 XXIV. Radiosensitizers, Radioprotectors and Bioreductive Drugs 176 XXV. Hyperthermia 178 XXVI. Radiation Carcinogenesis 179 XXVII. Heritable Effects of Radiation 182 XXVIII. Radiation Effects in the Developing Embryo and Fetus 183 XXIX. Radiation Protection 185 XXX. Molecular Techniques Used in Radiation and Cancer Biology 188 XXXI. Molecular Imaging 191 Page 6 6 Preface to the 2012 Edition In recognition of the critical need to develop new ways to promote education in the biologic basis of radiotherapy, the Radiation and Cancer Biology Committee of ASTRO appointed a subcommittee to develop a dynamic web-based educational tool for radiation oncologists to further their studies of radiation and cancer biology. The ASTRO Radiation/Cancer Biology Practice Examination and Study Guide, referred to as the “RCB Study Guide” for sake of brevity, is the product of these efforts. The RCB Study Guide was created specifically with a goal to stimulate active learning. The topics included in the ASTRO RCB Study Guide are based upon those listed by the American Board of Radiology (ABR) on their web site for the cognitive exam in radiation and cancer biology. Therefore, the RCB Study Guide should provide guidance both to residents in their preparation for the radiation oncology certification examination as well as to more senior radiation oncologists taking the examination to demonstrate the cognitive expertise in radiation and cancer biology that is required for maintenance of certification. It is suggested that users of the RCB Study Guide attempt to answer the questions in each section and then review the correct answers and explanations. It is anticipated that this approach will lead to a more complete understanding of each topic. References are included, with a hypertext link to the abstract and article, for topics that are not addressed fully in the major radiation biology textbooks cited in the RCB Study Guide. It should be noted that for the selection of references, an emphasis was placed on recent review articles that provide current and comprehensive information on a particular subject. Users of the 2012 RCB Study Guide are encouraged to also review the material presented in the 2010 and 2011 versions. This is of importance since a goal associated with creation of the RCB Study Guides is to address all major topics in radiation and cancer biology of relevance to radiation oncologists over a three year period since it is not possible to include material for every pertinent subject in any single year. Radiation and cancer biology are dynamic fields with new results published daily in the scientific literature. The goal for radiation oncologists is to acquire a solid base of knowledge in radiation and cancer biology during their training and to build upon that foundation during their careers through regular reading of the scientific literature as well as attendance at seminars and scientific conferences. The RCB Study Guides are designed to help radiation oncologists achieve this goal. It is hoped that by helping to provide radiation oncologists with a firm foundation in the biologic principles underlying the treatment of cancer with radiation, they will be able to offer more effective radiotherapy and achieve improved clinical outcomes for their patients. Finally, I would like to thank all of the current and former Associate Editors and Contributors who wrote and carefully reviewed the questions, explanations and references. Without your assistance, creation of the ASTRO Radiation and Cancer Biology Practice Examination and Study Guides would have not been possible. Barry S. Rosenstein, Ph.D. January, 2012 Page 7 7 Note on Protein and Gene Nomenclature The 2012 ASTRO Radiation/Cancer Biology Practice Examination and Study Guide uses the notation system for the name of each gene and protein encoded by that gene that was developed by the HUGO Gene Nomenclature Committee. The details for that system can be found at http://www.gene.ucl.ac.uk/nomenclature/. The guidelines for this system stipulate that gene symbols are italicized and designated by upper-case Latin letters or by a combination of upper-case letters and Arabic numerals. The protein encoded by the gene is given the same symbol as the gene, except that the letters are not italicized. Thus, the symbol for the gene mutated in people with the disease ataxia telangiectasia is ATM and the protein encoded by that gene is written as ATM. It should be noted that although the HUGO is widely used in scientific journals and textbooks, this system is rarely used for some proteins and genes. For these genes/proteins, the common symbol has been used in the exam/study guide, but the HUGO symbol is provided in parentheses the first time that the gene/protein is written in the question. For example, p53 is used in the exam/study guide rather than the official HUGO symbol for this gene, which is TP53. This is noted by indicating p53 (TP53) in the question or explanation. Page 8 8 QUESTIONS Page 9 9 I. Interaction of Radiation with Matter I-1) In terms of radiation-induced damage to mammalian cells, the most important radiolysis products of water are: A. aqueous solvated electrons B. hydrogen atoms C. superoxide radicals D. hydrogen peroxide E. hydroxyl radicals I-2) Which of the following statements concerning the different types of radiation- induced DNA damage is correct? A. For low LET radiation, the ratio of double- to single-strand breaks produced is approximately 20:1. B. In terms of cell killing, DNA-lipid crosslinks are considered the most deleterious form of DNA damage. C. High LET radiation is more likely to generate clustered DNA lesions (i.e. locally multiply damaged sites) than low LET radiation. D. Irradiation in the absence of oxygen enhances radiation toxicity. E. Decreasing the concentration of intracellular thiols protects against radiation toxicity. I-3) Biologic effects produced by X-rays are due primarily to: A. direct ionization of DNA molecules B. damage to critical cellular proteins C. direct ionization of the plasma membrane D. indirect damage done to DNA and the cell membrane E. lipid peroxidation I-4) Each of the following statements concerning radiation interactions with matter is true, EXCEPT: A. In the energy range generally used for radiation therapy, the Compton effect is the predominant interaction between photons and the matter through which they pass. B. Pair production occurs for photon energies greater than 1.02 MeV. C. Coherent scattering involves photon scatter, but without absorption. D. The probability of the photoelectric effect occurring is inversely proportional to the atomic number of the absorbing medium. E. An annihilation reaction occurs when an electron interacts with a positron to produce two 0.511 MeV photons. Page 10 10 I-5) The energy average LET (in keV/μm) for 14 MeV neutrons is approximately: A. 1 B. 10 C. 25 D. 100 E. 250 I-6) As the energy of a particulate type of radiation increases, the LET tends to: A. increase B. decrease C. first increase and then decrease D. first decrease and then increase E. remain unchanged I-7) Which of the following pairs of types of radiation and approximate LET (in units of keV/µm) is incorrect? A. 150 MeV protons – 100 keV/μm B. 1 GeV/nucleon Fe ions – 150 keV/μm C. 60 Co γ-rays – 0.2 keV/μm D. 2.5 MeV α-particles - 150 keV/μm E. 250 kV X-rays – 2 keV/μm I-8) The indirect effect for low LET radiation results from: A. characteristic X-rays produced by the photoelectric effect B. pair production C. hydroxyl radicals produced by the radiolysis of water D. ionizations generated within critical target molecules E. scavenging of free radical damage Page 11 11 II. Molecular Mechanisms of DNA Damage II-1) Which of the following DNA alterations is NOT generally associated with exposure to ionizing radiation? A. double strand breaks B. deletions C. microsatellite instability D. mutations E. pyrimidine dimers II-2) The type of ionizing radiation-induced DNA damage most likely to result in chromosome breakage and cell killing is a: A. single-strand break B. base damage C. DNA-protein crosslink D. clustered lesion E. ribose damage II-3) The number of DNA double-strand breaks produced per diploid mammalian cell by a γ-rays D0 (1-2 Gy) is approximately: A. 0.04 B. 1 C. 40 D. 1000 E. 400,000 II-4) Clustered DNA lesions are: A. composed only of base damages without any DNA strand breaks B. progressively more complex with increasing LET of the radiation C. thought to be easily and accurately repaired D. specifically detected using 53BP1 foci E. repaired by the mismatch repair pathway Page 12 12 II-5) The formation of DNA double strand breaks by increasing doses of radiation is best described as: A. Linear B. Quadratic C. Linear-Quadratic D. Parabolic E. Hyper-parabolic II-6 The formation of dicentric chromosomes by increasing doses of radiation is best described as: A. Linear B. Quadratic C. Linear-Quadratic D. Parabolic E. Hyper-parabolic Page 13 13 III. Molecular Mechanisms of DNA Repair III-1) The DNA double-strand break repair pathway that is largely “error-free” in normal cells is: A. translesion synthesis B. non-homologous end joining C. single-strand annealing D. homologous recombination E. post-replication repair III-2) Which of the following statements concerning the repair of radiation-induced DNA double strand breaks (DSB) is INCORRECT? A. The ataxia telangectasia mutated (ATM) protein is part of the initiation step in a signaling cascade that activates DNA strand break repair. B. Although the RAD50/MRE11/NBS1 (MRN) complex contributes to DNA repair, defects in either MRE11 or NBS1 do not result in a radiation sensitive phenotype. C. Inactivation of either BRCA1 or BRCA2 affects homologous recombination-dependent DSB repair. D. BRCA1 is required for the formation of RAD51 nuclear foci. E. In response to DNA-damaging agents, the RAD51 protein localizes in nuclear foci that represent sites of DNA repair. III-3) Which of the following statements concerning DNA double strand break repair is INCORRECT? A. MRE11 and RAD51 are recruited to the site of the damage. B. γ-H2AX is a marker for DNA double strand breaks and has been used to monitor repair. C. Typically, phosphorylation of H2AX is not observed for approximately 3-6 hours following irradiation. D. Phosphorylated H2AX occurs on DNA near the site of double strand breaks. E. γ-H2AX participates in recruitment of proteins that constitute the repair machinery. Page 14 14 III-4) Which of the following proteins moves to the sites of DNA double strand breaks in chromatin following irradiation? A. SOD2 B. NBS1 C. TGFβ1 D. DNA polα E. XPF III-5) Which DNA repair process is used to repair replication errors? A. nucleotide excision repair B. base excision repair C. single strand annealing D. mismatch repair E. transcription-coupled repair III-6) Mutations in the BRCA1-BRCA2-Rad51 pathway have been shown to be associated with all of the following EXCEPT: A. defective homologous recombination B. hypersensitivity to DNA cross-linking agents such as cisplatin C. impaired DNA replication D. compromised ability to repair DNA double strand breaks E. defective translesion DNA synthesis III-7) The primary mechanism for the repair of DNA double-strand breaks during the G1 phase of the cell cycle is: A. nucleotide excision repair B. homologous recombination repair C. base excision repair D. mismatch repair E. nonhomologous end-joining Page 15 15 III-8) For which of the following proteins has evidence NOT been obtained to demonstrate a role in non-homologous end-joining? A. Ku70 (XRCC6) B. ligase IV C. DNA-PKcs (PRKDC) D. XRCC4 E. RAD51 III-9) Which of the following enzymes phosphorylates H2AX to form γ-H2AX in response to formation of DNA double strand breaks? A. MDM2 B. DNA-PK (PRKDC) C. p53 (TP53) D. CHK2 (CHEK2) E. PGE-lipas III-10) DNA glycosylases: A. rejoin strand breaks B. act specifically on purine base damage C. remove damaged bases D. insert undamaged bases after the removal of damaged bases E. are found in mammalian cells but not in bacterial cells III-11) The ataxia telangiectasia mutated (ATM) gene plays an important role in cellular radiation response by encoding: A. a critical protein necessary for scavenging oxygen free radicals B. an enzyme that phosphorylates and activates proteins involved in DNA strand break repair and cell cycle checkpoint control C. a ligase that directly rejoins DNA strand breaks D. a protein whose primary function is to bind and sequester Bcl-2 (BCL2), thereby altering radiation-induced apoptosis E. a DNA polymerase responsible for repair replication Page 16 16 III-12) Which of the following statements concerning interstrand crosslinks (ICLs) is INCORRECT? A. Fanconi anaemia (FA) is a disorder that results in sensitivity to ICLs B. Homologous recombination-mediated repair of ICLs is promoted by the FA pathway. C. Structure-specific nucleases and translesion polymerases participate in the coordinated removal of the ICL and the resumption or completion of DNA replication. D. ICL-based chemotherapy is rarely used in treatment of cancer. E. The FANC-A and B proteins are important in the ICL response. Page 17 17 IV. Chromosome and Chromatid Damage IV-1) Concerning chromosomal aberrations, which of the following statements is INCORRECT? A. Fluorescent in situ hybridization is useful for the detection of stable chromosomal aberrations many years after irradiation. B. The formation of a ring chromosome could be lethal to a cell. C. The induction of chromatid breaks by X-rays follows a linear function of dose. D. Chromosome-type aberrations are produced when a cell is irradiated prior to DNA synthesis. E. Scoring dicentric chromosomes in peripheral blood lymphocytes can be used for detecting radiation exposures only for whole body doses exceeding 2 Gy. IV-2) An example of an asymmetrical chromosome-type aberration is a(n): A. sister chromatid exchange B. reciprocal translocation C. inversion D. chromatid break E. dicentric chromosome IV-3) Radiation-induced terminal chromatid deletions increase as a: A. linear function of dose B. quadratic function of dose C. linear-quadratic function of dose D. exponential function of dose E. sigmoidal function of dose IV-4) Measurement of which one of the following chromosomal aberrations in peripheral blood lymphocytes many years following a putative whole body radiation exposure would provide the best estimate of the dose received? A. reciprocal translocations B. rings C. dicentrics D. chromosome breaks E. chromatid breaks Page 18 18 IV-5) Which one of the following statements concerning chromosomal aberrations is correct? A. A dicentric is a two-hit chromatid-type aberration. B. An inversion is a single-hit chromosome-type aberration. C. An anaphase bridge is a two-hit chromosome-type aberration. D. A translocation is a two-hit chromosome-type aberration. E. A ring chromosome is a single-hit chromatid-type aberration. IV-6) Concerning chromosomal aberrations produced by low LET radiation exposure, which of the following statements is FALSE? A. The detection of aberrations in mitogenically-stimulated lymphocytes can prove useful in the estimation of radiation exposure. B. The dose response curve for the induction of dicentric chromosomes is linear-quadratic in shape. C. Chromatid-type aberrations are produced when cells are irradiated prior to DNA synthesis. D. The radiation dose-rate affects the chromosomal aberration frequency. E. Fluorescent in situ hybridization (FISH) facilitates the identification of translocations. Page 19 19 V. Mechanisms of Cell Death V-1) Which of the following statements concerning autophagy is INCORRECT? A. Autophagy may promote cancer development by allowing cancer cells to overcome metabolic stress (hypoxia, lack of nutrients). B. Autophagy is a non-physiologic process that is only induced when cells are exposed to genotoxic agents C. Autophagy may suppress tumor progression through degradation of oncogenic proteins. D. Autophagy is controlled by the Atg family of proteins, E. Many anticancer agents, including radiation, may induce autophagy, although whether autophagy enhances their antitumor properties or contributes to therapeutic resistance often remains unclear. V-2) Morphological and biochemical features of apoptosis include all of the following, EXCEPT: A. DNA cleavage B. cell shrinkage C. condensation of chromatin at the periphery of the nucleus D. requirement for ATP E. rupture of the plasma membrane V-3) Molecular pathways leading to apoptosis involve which of the following proteins? A. caspases B. serine proteases C. casein kinases D. catalase E. glycosylases V-4) Mitotic catastrophe following irradiation is a consequence of: A. the presence of chromosome aberrations that interfere with cell division B. G1 arrest C. a reduction in cellular ATP levels D. radiation-induced senescence E. inactivation of tumor suppressor genes Page 20 20 V-5) All of the following are typical characteristics of cells undergoing apoptosis, EXCEPT: A. presence of cells with double their normal diploid DNA content B. plasma membrane blebbing with exposure of phosphatidylserine on the outer plasma membrane C. release of cytochrome c from the mitochondrial intermembrane space D. endonucleolytic cleavage of nuclear DNA accompanied by the presence of condensed chromatin E. activation of death receptors at the plasma membrane, formation of death-inducing signaling complexes, and activation of caspase-8 V-6) All of the following features of radiation-induced mitotic catastrophe are true, EXCEPT that it: A. is the primary mechanism that leads to cell death in the majority of cell types and solid tumors B. can be associated with failure to complete a normal cell division, followed by progression through another G1 and S phase C. can be caused by loss of the G2 checkpoint and failure of cytokinesis leading to polyploidy and fragmented nuclei D. generally leads to a rapid cell death within several hours following irradiation E. is associated with reduced clonogenic survival, but may also be compatible with continuing cell viability for several cell divisions V-7) Which one of the following statements regarding the regulation of apoptosis is INCORRECT? A. The anti-apoptotic factors Bcl-2 (BCL2) and Bcl-XL (BCL2L1) inhibit release of cytochrome c from mitochondria. B. Cleaved BID binds to BAX and BAK, causing the release of cytochrome c from mitochondria. C. Pro-apoptotic BCL2 family members promote the release of cytochrome c. D. Apoptotic protease-activating factor 1 (APAF1) activates caspase 9, which in turn activates downstream caspases such as caspase 3, leading to apoptosis. E. XIAP plays a central role in the stimulation of apoptosis. Page 21 21 V-8) All of the following statements are true concerning sphingomyelinase, EXCEPT: A. It is activated in cells in response to high doses of irradiation. B. It induces the production of ceramide from sphingomyelin. C. It is involved in a signaling pathway that can lead to radiation-induced apoptosis. D. It can contribute to radiation-induced endothelial cell death. E. Its absence in endothelial cells results in a marked stimulation of radiation-induced apoptosis. V-9) Which one of the following proteins is involved in the induction of apoptosis following irradiation? A. p53 B. survivin C. EGFR D. PI-3K E. VEGF V-10) Radiation-induced bystander effects have been demonstrated in experiments where: A. cells were treated with several small doses of radiation rather than a larger single dose B. unirradiated cells were treated with medium taken from cell cultures previously exposed to low-LET radiation C. cells die following small single doses of ionizing radiation below approximately 0.2-0.3 Gy D. a low dose exposure protected cells against a subsequent high dose exposure E. effects are seen in cells up to 12 generations after radiation exposure Page 22 22 V-11) Which of the following statements regarding radiation-induced cell killing is INCORRECT? A. Morphological evidence of radiation damage can be seen immediately following irradiation in most cells. B. In some radiosensitive cell populations, cells can undergo an early interphase death within a few hours of irradiation. C. The cell membrane may be involved in triggering radiation-induced apoptosis in some cell types. D. Cycling cells in irradiated tissues may die due to mitotic catastrophe. E. Senescent or terminally-arrested cells die days to weeks post-radiation from necrosis. V-12) Which of the following cell types is particularly prone to apoptotic death following irradiation? A. dermal fibroblasts B. neurons C. astrocytes D. lymphocytes E. hepatocytes Page 23 23 VI. Cell and Tissue Survival Assays VI-1) The Trypan Blue dye exclusion assay measures: A. DNA integrity B. cell membrane integrity C. protein synthesis D. RNA expression E. colony forming ability VI-2) The spleen-colony assay: A. is used to measure hepatocyte radiosensitivity B. requires roughly three weeks to permit spleen cells to form colonies in recipient animals C. was used to demonstrate that the radiation survival curve for intestinal crypt cells was linear-quadratic in shape D. was used to determine the radiation survival curve for bone marrow stem cells E. requires that the recipient animal be given a sublethal dose of radiation VI-3 A major limitation of in-situ colony formation assays for normal tissues, such as the testes clonogenic assay developed by Withers and his collaborators, is that they: A. are not useful for doses less than roughly 5 Gy B. are not able to provide estimates of the D0 C. require explanting cells from the irradiated tissue D measure functional endpoints, not cell survival E. primarily reflect radiation response of vascular endothelial cells Page 24 24 VII. Models of Cell Survival VII-1 In a clonogenic assay, cells were plated and allowed to divide for 24 hours (approximately 2 doubling times), before irradiation. They were then grown in situ for colony formation. As a result, the colonies arose from a cluster of several cells rather than from single cells. This technique would produce which of the following artifacts in the resulting survival curve? A. decreased n B. increased D0 C. decreased D0 D. increased n E. decreased Dq VII-2) A cell line is characterized by an X-ray survival curve with a Dq of 4 Gy and D0 of 1.5 Gy. What is the single dose that will produce the same surviving fraction as two 5 Gy fractions delivered 6 hours apart? A. 1 Gy B. 3 Gy C. 6 Gy D. 10 Gy E. 14 Gy VII-3) Which of the following would be a typical D0 value for hypoxic mammalian cells exposed to an acute dose of X-rays? A. 0.2 Gy B. 1 Gy C. 4 Gy D. 8 Gy E. 20 Gy VII-4) The “gold standard” endpoint for assessing cellular radiosensitivity is: A. necrosis B. loss of reproductive integrity C. mitotic rate D. trypan blue uptake E. giant cell formation Page 25 25 VII-5) If a cell line’s D0 equals its D37, then n is equal to: A. 0 B. 0.37 C. 1 D. 5 E. 20 VII-6) The quasi-threshold dose Dq is an indicator of all of the following, EXCEPT: A. the ability of the cell to accumulate sublethal radiation damage B. the width of the shoulder region of the cell survival curve C. how much sparing will be obtained by dose fractionation D. the cell’s sublethal damage repair capacity E. the duration of S phase of the cell cycle VII-7) The total dose, delivered as multiple, small dose fractions, required to produce a 90% probability of local control for a tumor containing 10 8 clonogenic cells and exhibiting an effective D10 of 5 Gy is: A. 5 Gy B. 25 Gy C. 35 Gy D. 45 Gy E. 60 Gy VII-8) Concerning the linear-quadratic model to describe cell survival curves, all of the following statements are true, EXCEPT: A. The α/β ratio is the dose at which the linear and quadratic components of cell killing are equal. B. A cell survival curve for high LET radiation typically has a large α/β ratio. C. The α/β ratio for an intact normal tissue or tumor can be obtained from a series of multifraction experiments employing different doses per fraction. D. At low doses, cell killing results primarily from more than one ionization track producing damage in separate chromosomes E. A large α/β ratio indicates that cells are relatively insensitive to changes in dose per fraction. Page 26 26 VII-9) A 9 Gy dose of X-rays delivered in 1.8 Gy fractions results in a cell surviving fraction of 10 -3 . Assuming that cell killing can be represented by the equation -lnS=D/D0, what dose would be required to reduce survival to 10 -9 ? A. 13 Gy B. 18 Gy C. 27 Gy D. 36 Gy E. this value cannot be determined with the information provided VII-10) Which of the following statements concerning cell survival curve parameters is INCORRECT? A. The reciprocal of the D0 corresponds to the final slope of the cell survival curve. B. For X-rays, the β component of cell killing is dependent on the square of the dose. C. α is a measure of the survival curve slope for a highly fractionated or very low dose rate treatment. D. Dq is generally large for high LET radiation. E. The extrapolation number, n, is usually small for high LET radiation. VII-11) Radiation-induced cell killing depends on all of the following, EXCEPT the: A. dose rate B. LET of the radiation C. cell cycle phase D. partial pressure of oxygen in the tissue E. fraction of cells already killed VII-12) A survival curve characterized by a low α/β ratio suggests that: A. the cell population as a whole contains subsets of cells with differing radiosensitivities B. the cells are capable of accumulating sublethal damage C. cell killing is an exponential function of dose D. a fraction of the total cell population is hypoxic E. radiation kills a greater number of cells per Gy at high doses compared to low doses Page 27 27 VIII. Linear Energy Transfer VIII-1) The RBE for a high LET form of radiation is: A. dependent on the isoeffect level at which it is measured B. independent of particle energy C. similar for aerated compared with hypoxic cells D. similar for cell lines of different origins VIII-2) Modifying which one of the following treatment parameters would cause the largest change in tumor response for high LET irradiation? A. total dose B. use of a hypoxic cell sensitizer C. dose rate D. partial pressure of oxygen E. use of smaller-than-conventional fraction sizes VIII-3) The RBE of 3 MeV α-particles is: A. typically in the range of 1.5-2.0 B. dependent upon dose rate C. generally greater for early effects than late effects D. independent of endpoint E. greater for a single dose than for a fractionated protocol VIII-4) One of the main reasons for continued interest in high LET radiotherapy is: A. the relatively low energies required to produce excellent depth dose characteristics B. less dependence of cell killing on oxygenation status C. reduced cost compared with conventional low LET radiotherapy D. better control rates for rapidly, compared to slowly, growing tumors E. a greater dependence of cell killing on cell cycle position Page 28 28 IX. Oxygen Effect IX-1) Groups of otherwise identical mouse tumors are irradiated with the same total physical dose (in Gy) under the following sets of treatment conditions. In which case would the tumors be expected to be most resistant to cure by the treatment? A. 200 MeV protons while tumors are clamped to produce hypoxia B. 250 kVp X-rays while the mouse is breathing carbogen C. 200 MeV protons while the mouse is breathing carbogen D. 200 MeV protons while the mouse is breathing normal room air E. 290 MeV carbon ions while tumors are clamped to produce hypoxia IX-2) For mammalian cells, which of the following would yield the lowest OER? A. 250 kVp X-rays B. 270 MeV/µ carbon ions in the Bragg peak C. 10 MeV neutrons D. 160 MeV protons in the Bragg peak E. 270 MeV/µ carbon ions in the plateau region IX-3) Which one of the following would not increase the radiation sensitivity of aerobic cells but would increase the radiation sensitivity of severely hypoxic cells? A. pretreatment with bromodeoxyuridine for 48 hours B. concomitant treatment with 5-fluorodeoxyuridine C. post-irradiation treatment with pimonidazole for 5 hours D. concomitant treatment with nimorazole E. concomitant treatment with amifostine Page 29 29 IX-4) Each of the following statements concerning the influence of oxygen on radiosensitivity is true, EXCEPT: A. Oxygen must be present during or within milliseconds following irradiation to act as a radiosensitizer. B. The partial pressure of oxygen at the cellular level must be similar to that in arterial blood for radiosensitization to occur. C. In the presence of oxygen, the single dose of X-rays required to produce a given level of effect is 2-3 fold lower than if the irradiation occurred under hypoxic conditions. D. Oxygen is a potent hypoxic cell radiosensitizer. E. Oxygen diffuses only a finite distance in respiring tissue. IX-5) Which of the following statements concerning the role of oxygen in radiation response is correct? A. Full radiosensitization occurs if oxygen is added up to 10 seconds after irradiation. B. The oxygen concentration corresponding to radiosensitivity halfway between the fully oxygenated and fully hypoxic response (K-value) occurs at a pO2 of approximately 2-10 mm Hg. C. Similar levels of DNA damage are produced in hypoxic and normoxic cells D. The OER increases rapidly as the pO2 increases from 155 to 760 mm Hg. E. The OER increases with LET up to approximately 20 keV/μm and then decreases. IX-6) A tumor that has been rendered completely hypoxic by clamping is irradiated with a single X-ray dose of 18 Gy. The tumor cell surviving fraction at this dose is 0.05. When an otherwise comparable tumor is irradiated with the same dose, but in an air-breathing animal, the surviving fraction is 0.01. For the second animal, the estimated tumor hypoxic fraction would be: A. 0.1 B. 0.05 C. 0.1 D. 0.2 E. 0.5 Page 30 30 IX-7) Concerning the measurement of tumor oxygenation status, which of the following statements is TRUE? A. Cervical carcinoma patients whose tumors had pO2 values >10 mmHg exhibited poorer outcomes than patients whose tumors had lower pO2 values. B. Tumor oxygenation status is likely to remain constant throughout treatment. C. Patients whose hypoxic tumors reoxygenate only slowly or not at all during the course of treatment would benefit from the use of an accelerated treatment schedule. D. A limitation in the use of tumor oxygen status is that a drug infusion and tumor biopsy is required. E. Pimonidazole has proven useful for the detection of hypoxia in human tumors. Page 31 31 X. Repair at the Cellular Level X-1) For most cell types irradiated in vitro, repair of sublethal damage is complete in approximately: A. 1 min B. 15-20 min C. 2 hours D. 10 hours E. 1 day X-2) As dose rate decreases from 1 Gy/min to 1 Gy/hour, which one of the following best describes the changes that will occur in cell survival curve parameters in most cell types? A. final slope increases B. D0 increases C. n increases D. survival curve does not change X-3) If cells in culture are exposed to two equal radiation doses separated by 5 hours, rather than an acute irradiation with the combined dose, the greatest increase in surviving fraction would be manifested for: A. bone marrow stem cells irradiated with 250 kVp X-rays B. melanoma cells irradiated with 200 MeV protons C. melanoma cells irradiated with 3 MeV α-particles D. bone marrow stem cells irradiated with 3 MeV α-particles E. fibroblasts irradiated with 300 MeV/µ carbon ions X-4) The inverse dose rate effect: A. occurs for radiation exposures at very high dose rates (e.g., >10 Gy/min) B. induces the bystander effect C. is a result of potentially lethal damage repair during irradiation D. is manifest as an increase in cell killing with decreasing dose rate E. occurs only in non-proliferating cells such as neurons Page 32 32 X-5) When cells are maintained under suboptimal growth conditions for 6 hours after a single dose of X-rays, the cell surviving fraction is noted to increase. This is evidence for the: A. repair of sublethal damage B. redistribution of cells around the cell cycle C. repair of potentially lethal damage D. repair of base damages E. inverse dose rate effect X-6) An 8 Gy X-ray dose delivered at 1 Gy/hr is less toxic than the same dose delivered at 1 Gy/min primarily because: A. fewer free radicals are generated B. cell division occurs during exposure C. free radical scavenging takes place D. sublethal damage repair occurs during the irradiation E. chemical restitution is permitted X-7) The repair of sublethal damage: A. increases as the time between two X-ray dose fractions is decreased B. is of greater importance for cells exposed to high LET compared with low LET radiation C. has a time course similar to that for DNA double strand break rejoining D. occurs in vivo, but not in vitro E. is generally enhanced by holding cells under sub-optimal growth conditions after irradiation X-8) Which one of the following normal tissues would be expected to show the least amount of sparing when irradiated with X-rays at a low versus high dose rate? A. kidney B. lung C. spinal cord D. breast epithelium E. bone marrow Page 33 33 XI. Solid Tumor Assay Systems XI-1) The effect of a dose of radiation on the growth of experimental rodent tumors is most rigorously measured by: A. sacrificing the animals at a fixed time after irradiation and determining the weights of the treated versus control tumors B. sacrificing the animals at a fixed time after treatment and measuring the volumes of the treated versus control tumors C. making sequential measurements of tumor volumes for a fixed time interval after treatment and comparing the tumor volumes for treated and control animals on different days D. obtaining sequential measurements of the tumor volumes until each tumor reaches a predetermined volume and then calculating the growth delay produced by the treatment E. determining the times at which the treated and control tumors cause death of the animal XI-2 The lung colony assay: A. can be used to measure the survival of leukemia cells B. measures tumor cell proliferation rates C. is used to measure tumor growth delay D. measures the survival of type II pneumocytes after lung irradiation E. is a clonogenic assay used for determining tumor cell survival XI-3) For the endpoint dilution assay technique, if the control TD50 is 2 cells and the TD50 following irradiation is 8 cells, then the normalized surviving fraction for the irradiated cells is: A. 0.004 B. 0.04 C. 0.16 D. 0.25 E. 0.80 Page 34 34 XI-4 Which of the following could not be used as a host for a human tumor xenograft? A. nude mice B. SCID mice C. nude rats D. hamster cheek pouch E. C3H mice carrying MMTV Page 35 35 XII. Tumor Microenvironment XII-1) Which of the following is NOT a way in which a hypoxic microenvironment can influence a tumor cell population? A. selection for intrinsically radioresistant cells B. facilitating genomic instability C. selection for cells with decreased apoptotic potential D. inhibition of cell proliferation E. increased potential for metastases XII-2) Which of the following is NOT a characteristic of oxygen and its influence on radiotherapy? A. Repair of damage between radiation fractions reduces overall radiation effects, whereas reoxygenation of hypoxic cells between fractions increases overall radiation effects in tumors. B. Reoxygenation of hypoxic cells between fractions is limited to ‘acutely’ hypoxic cells. C. Tumor hypoxia would be expected to have a larger impact for a treatment performed with a single fraction compared with a standard fractionated protocol. D. Hypoxia can be found in tumours smaller than 1 cm. XII-3) Oxygen measurements in patients have demonstrated which of the following? A. Oxygenation levels are similar across patients with similar tumor types. B. Extremely low levels of oxygen, such as those required to cause full resistance to radiation, are rarely observed. C. Tumor hypoxia is found exclusively in the central core of large tumors. D. Oxygenation levels are relatively stable over time. E. Some tumors contain no detectable hypoxic cells. Page 36 36 XII-4) All of the following are the specific potential advantages afforded to patients by functional and molecular imaging techniques compared to current clinical imaging modalities, EXCEPT: A. minimal invasiveness B. improved assessment of treatment efficacy C. better clinical staging and stratification of risk D. near-simultaneous, quantitative assessment of multiple biochemical processes E. more efficient drug discovery and validation of drug targeting XII-5) All of the following statements regarding the interaction of tumor cells and their associated stroma are true, EXCEPT: A. Cancer cells can alter their adjacent stroma to form a supportive environment for tumor progression. B. Stromal cells tend to be more genetically unstable than cancer cells and are therefore more likely to become drug resistant. C. Cancer cells modify their stromal microenvironment through the production of a range of growth factors and proteases. D. Tumor stroma can have a direct role in tumorigenesis through oncogenic signaling and mutagenesis. E. Stromal cells can enhance tumor progression by secreting growth factors and pro-migratory extracellular-matrix components. XII-6) Which of the following agents has NOT been found to be useful for measuring human tumor hypoxia? A. [ 18 F]- 2-fluoro-2-deoxy-D-glucose (FDG), B. pimonidazole C. [ 18 F]Azomycin Arabinoside (FAZA) D. [ 64 Cu]- Copper dithiosemicarbazone (ATSM) E. [ 18 F]Fluoromisonidazole (FMISO) Page 37 37 XII-7) Pre-treatment correction of anemia with either blood transfusion or erythropoietin administration may be considered in cancer patients. This is based, in part, on which one of the following observations? A. In a German/Swiss, multicenter, randomized, clinical trial, advanced head and neck cancer patients receiving epoetin alfa achieved higher hemoglobin concentrations and longer progression-free survival than patients receiving placebo. B. Anemia correction has been shown to improve quality of life by reducing fatigue. C. In the Breast Cancer Erythropoietin Survival Trial (BEST), patients with metastatic breast cancer demonstrated improved overall survival when treated with epoetin alpha compared to placebo. D. The activation of hypoxic signaling pathways, including HIF-1α, is associated with resistance to therapy and depends critically on hemoglobin levels. E. Erythropoietin has been successfully administered to victims of radiation accidents receiving doses of 8 Gy or higher. XII-8) Which one of the following statements describing tumor vasculature is correct? A. Tumor blood vessels are hyper-permeable, tortuous, and feature haphazard patterns of interconnection, resulting in spatial and temporal heterogeneity of tumor blood flow. B. Tumor blood vessels are hypopermeable, irregular, and densely invested with pericytes resulting in poor diffusion of chemotherapeutics into tumor parenchyma. C. Tumor blood vessels are indistinguishable from normal blood vessels ultrastructurally, however compression caused by proliferating tumor cells leads to vessel collapse and compromised blood flow. D. Tumor blood vessels are dilated, tortuous and have uniformly thick basement membranes, resulting in limited accessibility of chemotherapy agents. E. Tumor blood vessel organization resembles that of normal vessels, however tumor vessels differ ultrastructurally, resulting in poor function and heterogeneous blood flow. Page 38 38 XII-9) The regulation of hypoxia-inducible factor-1α (HIF-1α) (HIF1A) by oxygen concentration is best described by which of the following statements? A. Under hypoxic conditions, HIF-1α transcription and translation are up- regulated, causing the protein to translocate from the cytosol to the nucleus. B. Under aerobic conditions, the HIF-1α heme moiety becomes oxygenated. This drives a conformational change in the protein limiting DNA binding and thereby preventing up-regulation of target genes. C. Under hypoxic conditions, HIF-1α is activated by bioreduction, thereby promoting the up-regulation of target genes. D. Under hypoxic conditions, the HIF-1α heme moiety becomes deoxygenated. This causes a conformational change in the protein, enhancing DNA binding and thereby promoting up-regulation of target genes. E. Under aerobic conditions, HIF-1α is hydroxylated by HIF prolyl hydroxylases. This targets the protein for ubiquitination and subsequent proteosomal degradation, thereby preventing the up-regulation of target genes. XII-10) All of the following statements concerning tumor hypoxia are correct, EXCEPT: A. Hypoxia-inducible factors primarily regulate oxygen concentration in tumors, but exert little or no control over tumor invasiveness and metastasis. B. Hypoxia signaling is involved in the regulation of the epithelial– mesenchymal transition, invasion, intravasation, survival of circulating tumor cells, extravasation, formation of the premetastatic niche and growth from micrometastais to macrometastasis C. A possible negative aspect of anti-angiogenic therapy is that it may increase tumor hypoxia, thereby increasing the harmful consequences resulting from this state. D. It has been suggested that the repair of the abnormal structure and function of tumor vessels through ‘sustained vessel normalization’ improves oxygen supply, reduces metastasis and might offer novel therapeutic opportunities. E. Hypoxia-inducible factors are regulated by oxygen sensing prolyl hydroxylases. Page 39 39 XII-11) Which of the following statements about hypoxia-inducible factors (HIFs) is INCORRECT? A. HIFs bind to DNA as heterodimers composed of alpha- and beta- subunits, both of which are oxygen sensitive. B. HIFs facilitate oxygen delivery by regulating expression of genes whose products are associated with angiogenesis and erythropoiesis. C. HIFs facilitate adaptation to oxygen deprivation by regulating gene expression for glucose uptake and cellular metabolism. D. HIFs regulate gene expression for cellular proliferation and apoptosis. E. HIF activation correlates with metastasis in multiple tumors. XII-12) Which of the following statements about tumor hypoxia is INCORRECT? A. The presence of intratumoral hypoxia is a positive prognostic indicator. B. Hypoxia can drive the metastatic phenotype as a result of genetic instability, increased angiogenesis, decreased apoptosis and upregulation of a of genes involved in the metastatic cascade. C. Hypoxic cells can acquire a mutator phenotype that is characterized by decreased DNA repair, an increased mutation rate and increased chromosomal instability. D. Defects in homologous recombination and mismatch repair have been documented in chronically hypoxic tumor cells. E. High LET radiations such as alpha particles have no better killing of hypoxic than oxic regions of tumors. Page 40 40 XIII. Cell and Tissue Kinetics XIII-1) Following exposure to ionizing radiation, cells that lack functional p53 are most likely to arrest in which phase of the cell cycle? A. G1 B. S C. G2 D. M E. The cells will not arrest. XIII-2) Radiation-induced G1 arrest involves which one of the following sequences of protein changes? A. FAS ligand binding, caspase 9 activation, mitochondrial breakdown B. ATM autophosphorylation, p53 (TP53) phosphorylation, p21 (CDKN1A) upregulation C. Cytochrome c release, caspase 8 activation, lamin degradation D. PI3K acetylation, p53 ubiquitination, Bcl-2 (BCL2) degradation E. NBS1 phosphorylation, ATM methylation, BAX upregulation XIII-3) The phase of the cell cycle that exhibits the greatest resistance to ionizing radiation is: A. M B. G1 C. Early S D. Late S E. G2 XIII-4) Most proliferating human tumor cells in vivo have cell cycle times in the range of: A. 6-24 hours B. 1-5 days C. 5-25 days D. 4-8 weeks E. 10-12 weeks Page 41 41 XIII-5) Assuming that all cells in a tumor are proliferating, if the cell cycle time is 3 days, the labeling index is 0.2 and λ is 0.7, what is the approximate duration of S phase? A. 1 hr B. 5 hr C. 10 hr D. 20 hr E. 50 hr XIII-6) The time between the 50% points for labeled mitotic cells in the first peak of the percent labeled mitoses curve measures the length of: A. M B. G1 C. S D. G2 E. Tc XIII-7) If an asynchronous population of 2 x 10 7 cells has the following cell cycle parameters: TM = 1 hr; TG1 = 11 hr; TS = 5 hr; and TG2 = 3 hr and the growth fraction is 0.5, approximately how many cells would be expected to be in S phase? A. 2.5 x 10 5 B. 10 6 C. 2.5 x 10 6 D. 5 x 10 6 E. 10 7 XIII-8) Which one of the following statements concerning tumor cell cycle kinetic parameters is true? A. Values for TS and MI are required in order to calculate Tpot. B. Tpot can be estimated from a single tumor biopsy taken approximately 48 hours after the administration of either bromo- or iododeoxyuridine. C. TS can be estimated from the time required for the earliest S phase cells in a population at the time of labeling to progress into G2. D. Randomized, prospective clinical trials have all confirmed that tumors with short Tpot values are the best candidates for accelerated fractionation. E. Tumors with long Tpot’s are most likely to benefit from hyperfractionation. Page 42 42 XIII-9) Which pair of cell cycle phase and active CDK or enhanced cyclin level is INCORRECT? A. G1 - CDK1 B. S - CDK2 C. G1 - CDK4 D. G2 - cyclin B E. G1 - cyclin D XIII-10) The DNA content of G0 cells usually equals that of: A. S phase cells B. G1 cells C. G2 cells D. mitotic cells E. tetraploid cells Page 43 43 XIV. Molecular Signaling XIV-1) Which of the following is NOT a component of, or a target for, the RAS signaling pathway? A. mitogen-activated protein kinase (MAPK) pathway B. ATM activation C. phosphoinositol 3 kinase (PI3K) pathway D. inherent radiosensitivity E. farnesyltransferase inhibitors XIV-2) Which of the following statements does NOT correctly describe the ERBB family of kinases? A. ERBB2/HER-2 is over-expressed in fewer than approximately 5% of metastatic breast cancers. B. ERBB2/HER-2 is the receptor that binds Herceptin. C. EGF receptor kinase inhibition can be achieved using antibodies or small molecule kinase inhibitors. D. ERBB receptors activate signaling through PI3 kinase and protein kinase B/AKT to enhance cell survival. E. EGF receptor mutations can be found in malignant gliomas and lung cancer XIV-3) Which of the following statements does NOT describe a role attributed to the mammalian target of rapamycin (mTOR) pathway in cancer and cancer therapy? A. The PI3K–AKT pathway activates mTOR. B. Rapamycin analogues that inhibit mTOR may be useful additions to radiotherapy. C. mTOR mainly functions to reduce proliferation. D. Rapamycin analogues slow the growth of tumors in animal models E. mTOR suppresses autophagy. Page 44 44 XIV-4) Which of the following phrases describes an important function of wild type p53 (TP53) protein? A. inhibition of downstream proteins necessary for cell cycle arrest following radiation-induced DNA damage B. detoxification of reactive oxygen species generated by radiation by sequestering reduced glutathione in the nucleus C. upregulation of proteins such as BAX and PUMA to promote radiation- induced apoptosis D. phosphorylation of the histone variant, H2AX E. activation of ATM . XIV-5) MDM2 is thought to: A. act as an E3 ligase that drives the degradation of p53 (TP53) protein B. participate as a partner with p53 in binding to the cyclin dependent kinases and initiating cell cycle arrest C. bind directly to p21 when over-expressed in p53 deficient cells thereby initiating cell cycle arrest D. increase the overall expression of p53 by binding to the p53 promoter and increasing its transcription E. enhance the stability of p53 XIV-6). Which one of the following is a proteasome inhibitor? A. Rituxan B. Taxol C. Bortezomib D. Celecoxib E. Erbitux XIV-7) Farnesyltransferase inhibitors (FTIs) were originally developed to target: A. BAX B. Caspase 3 C. RAS D. TNF-α E. DNA-PKcs Page 45 45 XIV-8) Ionizing radiation has been shown to up-regulate many genes, depending both on radiation dose and time after irradiation. Because of this, there is interest in developing gene therapy vectors containing suicide genes with radiation-inducible gene promoters in order to increase the toxicity of radiation locally at the tumor site. Which one of the following genes contains a promoter that has been used for this purpose? A. EGR1 B. Actin C. Survivin D. Caspase 8 E. Ku-80 XIV-9) Which one of the following is a correct pairing of a small molecule inhibitor of intracellular signaling and its target protein? A. Gleevec - p53 (TP53) B. Iressa – EGFR C. Wortmanin - MDM2 D. Gemcitabine - PI3K E. Herceptin - MAPK XIV-10) Which of the following processes is NOT a component of the cascade of events that comprises a signaling pathway? A. receptor-ligand interaction B. transfection C. tyrosine kinase phosphorylation D. signal transduction E. ubiquitination XIV-11) Which of the following statements concerning RAS is INCORRECT? A. RAS proteins are small GTPases . B. Genes encoding RAS proteins are rarely found to be mutated in human cancers. C. Guanine nucleotide exchange factors (GEFs) and GTPase-activating proteins (GAPs) control the GDP–GTP cycling of RAS proteins. D. GEFs and GAPs are have been found to be deregulated in human cancers E. The first genes identified as encoding RAS proteins were HRAS and KRAS. Page 46 46 XIV-12) Which of the following statements concerning ERBB inhibitors is INCORRECT? A. Humanized antibodies directed against the extracellular domain of EGFR or ERBB2 represent forms of ERBB inhibitors in clinical use. B. ERBB-directed tyrosine-kinase inhibitors (TKIs) and antibodies have been shown to inhibit tumor cell proliferation in vitro. C. Small-molecule TKIs that compete with ATP in the tyrosine-kinase domain of the ERBB receptor have been developed. D. ERBB inhibitors generally exhibit greater clinical efficacy than predicted based upon pre-clinical studies. E. New ERBB inhibitors continue to be developed as therapeutic agents. Page 47 47 XV. Cancer XV-1) The proto-oncogene that appears to be involved in radiation-induced thyroid cancer is: A. RET B. ATM C. ERB-B D. SIS E. C-MYC XV-2) For which of the following genes has evidence been obtained indicating that certain mutations present in the gene may serve as prognostic genetic markers for tumor resistance to radiation treatment? A. XRCC4 B. BRCA2 C. RAS D. ATM E. XRCC1 XV-3) Which of the following statements concerning epigenetic changes commonly found in cancers is INCORRECT? A. DNA hypermethylation of immunity-related genes in some tumor cells allows them to escape recognition by the host’s immune system. B. Certain tumor suppressor genes show hypermethylation in cancer. C. Oncogenes generally exhibit hypermethylation in cancer. D. “Epigenetic therapy” is currently being tested in clinical trials for certain malignancies. E. Histone deacetylation and DNA methylation are implicated in gene silencing. Page 48 48 XV-4) Which of the following statements concerning cancer vaccines is TRUE? A. Most tumor cells have all of the necessary characteristics to elicit a strong immune response. B. The idea that the immune system can be deliberately manipulated to both recognize and eradicate cancer is a recent idea. C. Only T-cells are capable of being activated to produce an immune response against a tumor. D. Gene therapy of tumor cells with immunomodulatory cytokine genes does not enhance the efficacy of tumor vaccines. E. Sipuleucel-T (Provenge) is a form of immunotherapy approved for treatment of prostate cancer. XV-5) True statements concerning possible applications of nanotechnology in cancer diagnosis and therapy include all of the following, EXCEPT: A. Semiconductor nanocrystals called quantum dots each emit visible light of a different wavelength based upon their size. These can be used as tags for different biomolecules whose activities can be detected simultaneously. B. One major limitation of nanotechnology approaches to cancer imaging or therapy is the difficulty of the nanoprobe in crossing biological or biophysical barriers. C. Nanoscale gold-plated beads of glass called nanoshells can be targeted to cancer cells with antibodies, and then heated using an external infrared source to provide highly localized hyperthermia. D. New nanoparticulate formulations allow simultaneous imaging and therapy (“theranostics”). E. Carbon nanotubes have been used for radiotherapy. XV-6) Loss of p16 INK4a expression may alter the cellular response to radiation- induced damage because this protein normally: A. is necessary for non-homologous end joining to proceed B. is required to enhance Bcl-2 function C. inhibits the CDK4/6:cyclin D complex D. is necessary to bind to FAS ligand E. stimulates CHK2 Page 49 49 XV-7) Which of the following is NOT an oncogene? A. MYCN (N-myc) B. HER2 C. MET D. PTEN E. RET XV-8) Human cancer cells are commonly defective in which of the following proteins normally involved in G1/S checkpoint control in response to ionizing radiation? A. CDK4 B. p53 (TP53) C. cyclin B D. Myc E. ATM XV-9) Increased expression of which of the following proteins is important in the immortalization step of carcinogenesis? A. telomerase B. cyclin B C. p53 (Tp53) D. DNA-PKcs E. ligase IV XV-10) Which of the following statements is TRUE regarding the retinoblastoma protein RB (RB1)? A. RB is rarely mutated in human cancers. B. When RB and E2F are complexed together, cells are stimulated to progress from G1 into S phase. C. CDK1/cyclin B phosphorylates RB. D. Hyper-phosphorylation of RB causes a conformational change in this protein resulting in its detachment from E2F. E. Elevated levels of cyclin D protein are associated with mutated RB. Page 50 50 XVI. Total Body Irradiation XVI-1) Which of the following organs is most at risk of developing life-threatening late complications after 12 Gy of total body irradiation prior to bone marrow transplantation? A. GI tract B. Kidney C. Liver D. Skin E. Pancreas XVI-2) A radiation accident exposes an individual to a whole body dose of about 4 Gy. Which of the following statements regarding the likely clinical symptoms is INCORRECT? A. The accident victim will develop nausea and vomiting. B. The person will experience immediate and prolonged diarrhea, dehydration and weight loss. C. Following the prodromal syndrome period, the victim will be largely symptom-free for approximately 3 weeks. D. The latent period will be followed by the onset of chills, fatigue and declining blood cell counts. E. The victim will not become anemic. XVI-3) People exposed to acute, whole body radiation, can be treated in different ways depending on the total dose received. Which of the following dose treatment option pairs is correct? A. 0.5-1 Gy : transfusion with packed red blood cells B. 1-2 Gy : immediate platelet infusion C. 2-5 Gy : bone marrow transplant D. 5-10 Gy : ambulatory monitoring E. 10-20 Gy : there are no treatment options other than providing comfort Page 51 51 XVI-4) Prussian blue is used therapeutically to prevent uptake of which of the following radionuclides in the event of a nuclear accident? A. I in the thyroid B. Pu in bone C. Cs in the intestines D. Am in bone E. Rn in lung XVI-5) A terrorist explodes a “dirty bomb” that contaminates your hospital with 137 Cs. Which one of the following emergency responses is inappropriate? A. Isolate, block off, and guard the contaminated area to prevent unauthorized entry. B. Treat all life threatening physical injuries in both contaminated and uncontaminated victims of the accident. C. Notify appropriate local, state, and federal agencies. D. Monitor everyone leaving the area for radioactive contamination; decontaminate when needed. E. Administer potassium iodide to all exposed individuals within 1 hour. XVI-6) Which pair of effects and total body acute dose is incorrect? A. cerebrovascular syndrome – 50 Gy B. hypotension – 1 Gy C. hematopoietic syndrome – 4 Gy D. gastrointestinal syndrome – 12 Gy E. nausea – 2 Gy XVI-7) A worker has been exposed accidentally to a total body dose in the range of 8-10 Gy. The full benefit of current radiation treatment protocols is available, including multiple courses of antibiotics, stem cell transfusions, cytokine supplements and other supportive measures. Nevertheless, the worker dies one year later. Which of the following is most likely to have been the cause of death? A. cerebrovascular syndrome B. gastrointestinal syndrome C. radiation dermatitis D. radiation-induced lung fibrosis E. hematopoietic syndrome Page 52 52 XVI-8) Following a large-scale radiological event, the current lack of rapid assays to measure radiation dose may mean that the presence and extent of acute symptoms could be the only way to triage victims into different treatment groups. In such a scenario, which of the following statements is true? A. Granulocyte numbers will fall steadily and can be used to assess dose. B. Lack of vomiting until 12 hours after the exposure suggests a radiation dose has been received that is not life-threatening. C. The degree of erythema can serve as a biodosimeter. D. After receiving a whole body dose of 3 Gy, anemia will be detectable within 24 hours following irradiation. E. For whole body doses in the range of 2-5 Gy, prodromal symptoms will be continuous and ultimately merge with the characteristic symptoms of the whole body radiation syndromes XVI-9) Total body irradiation (TBI) prior to hematopoietic stem cell transplant (HSCT) is often given with doses of 12 Gy or more. Which of the following statements explains why death from the gastrointestinal syndrome is not inevitable after TBI prior to HSCT? A. Patients undergoing HSCT are healthy and resistant to lethal effects of myeloablative TBI. B. During TBI, the entire gastrointestinal tract is shielded and this prevents the lethal gastrointestinal syndrome. C. TBI is fractionated or delivered at low dose rate and this increases tolerance of the gastrointestinal tract. D. Hematopoietic stem cells repopulate the gastrointestinal crypts and prevent the gastrointestinal syndrome from occurring. XVI-10) A nuclear submarine’s reactor malfunctions and acutely exposes the entire crew of 100 men to the same uniform dose of ionizing radiation. The only medical officer has no access to antibiotics. It takes two months for the disabled submarine to safely return to a military base, at which time only 10 men have survived. Assuming radiation was the cause of all deaths, what is the most likely approximate dose the crew received? A. 1 Gy B. 3 Gy C. 6 Gy D. 12 Gy E. 50 Gy Page 53 53 XVI-11) A patient is undergoing a reduced intensity allogeneic stem cell transplant for acute myelogenous leukemia. After receiving a single fraction of 2 Gy of total body irradiation, the hematologist realizes that the stem cell product is an inappropriate immunologic match for transplant and will inevitably lead to graft rejection. Different courses of action may then be taken, but which of the following would be considered to be INAPPROPRIATE? A. Discharge the patient home, with a follow-up appointment in 4 days. B. Admit the patient to a low acuity medical-surgical unit for antiemetics and observation. C. Admit the patient to an isolation room on the transplant ward and give the mismatched stem cell product. D. Admit the patient to an isolation room on the transplant ward and immediately initiate a search for an alternate suitable donor. E. Admit the patient to an isolation room on the transplant ward and begin collecting the hematopoietic stem cells in preparation for an autologous transplant. XVI-12) During a mass radiologic terrorism event, if only one laboratory test could be performed to assess for radiation exposure, which would be most valuable? A. Serum TGF-beta B. Serum creatnine C. Absolute neutrophil count D. Absolute lymphocyte count E. Serum IL-1 XVI-13) During a mass radiologic terrorism in which a truck bomb is exploded in a crowded urban area that results in dispersal of a low level of radioisotopes, victims that present with hypotension should be: A. immediately triaged to receive supportive care B. assessed and treated for trauma C. sent for neuroimaging to assess for cerebrovascular syndrome D. considered for bone marrow transplant E. be treated immediately with CSFs Page 54 54 XVII. Clinically Relevant Normal Tissue Responses to Radiation XVII-1) With regard to radiation-induced changes in kidney morphology following renal irradiation, all of the following can occur, EXCEPT: A. mesangiolysis and glomerulosclerosis B. tubular changes, including tubulolysis and atrophy C. pathognomonic features specifically characteristic of radiation-induced damage D. glomerulosclerosis and tubulointerstitial fibrosis E. morphologic changes in all components of the nephron XVII-2) With regard to radiation-induced brain injury, which of the following statements is FALSE? A. Brain irradiation can lead to an acute increase in blood-brain barrier leakiness and edema. B. Gray matter necrosis often occurs following brain irradiation. C. Brain irradiation leads to an acute burst in apoptotic cell death. D. Radiation-induced tissue changes are more frequently observed in AVM patients than tumor patients. E. Structural alterations noted following brain irradiation include demyelination and vasculopathy. XVII-3) All of the following statements regarding the acute and chronic side effects of irradiation to the central nervous system are true, EXCEPT: A. White matter necrosis is typically observed within 3 weeks following whole brain irradiation. B. Transient demyelination with Lhermitte’s syndrome after spinal cord irradiation represents an early delayed injury. C. Early delayed injury can involve transient demyelination with somnolence after brain irradiation. D. Long-term survivors of brain irradiation can exhibit cognitive impairment six months or more after brain irradiation. E. The severity of radiation-induced cognitive impairment is dependent on the patient’s age at the time of irradiation. Page 55 55 XVII-4) Which of the following statements most appropriately describes the response of human skin to fractionated radiotherapy? A. There is a large volume effect at low doses. B. The onset of moist desquamation is generally seen within 7 days from the initiation of treatment. C. Epilation is observed at doses similar to those that cause an early transient erythema. D. The latency period prior to the onset of moist desquamation is independent of dose. E. The severity of erythema and moist desquamation predict for the severity of late effects. XVII-5) Which one of the following strategies to reduce the severity of radiation- induced xerostomia is INCORRECT? A. use of IMRT to reduce the total volume of salivary gland tissue included in the treatment volume B. use of saliva substitutes and sialogogues as palliative agents C. stringent dental and oral hygiene D. administration of amifostine following the completion of radiation therapy XVII-6) Which of the following factors does NOT affect the incidence of osteoradionecrosis following radiation therapy of head and neck tumors? A. total radiation dose B. age at the time of radiation C. inclusion of marrow sinusoids in the field D. tooth extraction and/or dental disease E. site of primary tumor Page 56 56 XVII-7) Concerning radiation-induced damage to the eye, which of the following is FALSE? A. Fraction size is a major factor in the development of optic nerve lesions, with lesion incidence increasing from 5% to 40% for fraction sizes of 1 and 1.9 Gy, respectively. B. Conjunctivitis sicca (dry-eye syndrome) has a 100% incidence in patients receiving a total dose >55 Gy. C. The threshold dose for radiation-induced cataracts is less than 1 Gy (single dose). D. The threshold dose for radiation retinopathy is 40 Gy, increasing to a probability of 100% for a total dose of 70 Gy. E. Radiation-induced cataracts occur in the majority of children under the age of 5 years who receive a fractionated ocular dose of 0.5 Gy. XVII-8) Which one of the following statements regarding radiation-induced heart disease (RIHD) is FALSE? A. The α/β ratio for pericarditis is approximately 8 Gy. B. Endothelial cells lining the cardiovascular capillaries are considered to be radiosensitive whereas cardiac myocytes are relatively radioresistant. C. Fraction size, total dose and volume are all factors that can affect the tolerance dose for RIHD. D. Patients <20 years and >60 years of age have an increased risk of developing RIHD. E. Anthracyclines combined with radiation enhance cardiotoxicity compared to radiation alone. XVII-9) Which of the following statements is FALSE concerning effects of radiation on the testes? A. Permanent impairment of testicular endocrine function may occur following a testicular dose in the 20-30 Gy range. B. Fractionation increases the tolerance dose for sterility. C. A total dose of 1 Gy to the testes will cause transient sterility, lasting about 1 year. D. The most radiosensitive cells in the testes are the Type B spermatogonia. E. Despite long periods of azospermia, return of fertility is possible. Page 57 57 XVII-10) With regard to retreatment tolerance of normal tissues, which of the following statements is FALSE? A. Radiation-induced acute skin damage recovers well with restoration of almost full radiation tolerance. B. The spinal cord is capable of moderate long-term recovery. C. The kidney shows complete recovery from late functional damage. D. The lung exhibits retreatment tolerance for both lung fibrosis and pneumonitis. XVII-11) Which of the following statements regarding radiation-induced oral mucositis is true? A. Epithelial breakdown precedes endothelial cell loss. B. The probability of developing consequential late effects after radiation therapy for head and neck tumors does not vary with overall treatment time. C. The early stage of oral mucositis is associated with an acute neutrophil infiltrate. D. The incidence of oral mucositis is decreased by using accelerated hypofractionation. E. Oral mucositis can be prevented by using pentoxyfiline prior to therapy. XVII-12) Which of the following is NOT a common feature of delayed (chronic) intestinal radiation injury? A. irregular intestinal crypts B. hypertrophy of the mucosa C. dilated lymphatics D. submucosal fibrosis E. vascular sclerosis XVII-13) A patient was previously treated with radiation therapy to the spinal cord (44 Gy in 2 Gy fractions) and 3 years later presents with a metastasis causing spinal cord compression within the previously irradiated field. Based on data from non-human primates, what is the roughly the risk of myeloparesis by retreating this area to 50 Gy in 25 fractions? A. 100% B. 25-50% C. 1-10% D. 0.1-1% E. 0% Page 58 58 XVIII. Mechanisms of Normal Tissue Radiation Responses XVIII-1) Which of the following statements concerning radiation-induced damage to the microvasculature is INCORRECT? A. Classic pathologic features of radiation damage to blood vessels include telangectasia, “sausaging” and blebbing. B. Historically, vascular endothelial cells are considered the target cells for vascular injury, however, damage to adventitial fibroblasts and medial smooth muscle cells may also play significant roles. C. The order of radiosensitivity (from sensitive to resistant) of blood vessel is: capillaries; arterioles; small-medium arteries; large arteries. D. Due to the slow turnover kinetics of vascular endothelial cells, subcellular changes in these cells are generally not seen until approximately 6-12 months after irradiation. E. Bone marrow transplantation can result in pulmonary and hepatic veno- occlusive disease. XVIII-2) Which one of the following statements is INCORRECT concerning radiation- induced damage to the lung? A. Combined modality therapy with a variety of chemotherapeutic agents may enhance radiation-induced lung injury. B. Likely target cells for lung injury are endothelial cells of the alveolar capillaries and type II pneumocytes. C. Despite steep dose response curves, lung tolerance doses have been difficult to define because of large individual variations, observed both clinically and experimentally. D. For partial lung irradiation, both the volume and the location of the field affect the probability of developing pulmonary complications. E. Studies have shown that elevated plasma levels of TGFβ during and after treatment are independent negative indicators for the development of symptomatic lung damage. Page 59 59 XVIII-3) Which of the following statements is true regarding radiation-induced liver disease (RILD)? A. Hepatocytes are the principal radiobiological target. B. Liver is a flexible type tissue, so volume does not affect tolerance. C. High plasma levels of TGFβ predict for a decreased probability of developing RILD. D. Fraction size is not a factor in tolerance to RILD. E. In the bone marrow transplant setting, symptoms of RILD are observable within 3 months after treatment completion. XVIII-4) The relationship between total dose and the volume of tissue irradiated can be explained by all of the following statements, EXCEPT: A. In the skin and spinal cord, migration of surviving clonogenic cells from neighboring functional subunits (FSUs) can explain the high tolerance dose when irradiating small volumes of tissue. B. FSUs of the skin are arranged in series, and show a linear tolerance dose-volume relationship at low doses. C. The radioresponse of the spinal cord is consistent with a serial arrangement of FSU’s. D. For short lengths of spinal cord (<1 cm), an increase in field size is associated with a marked increase in the probability of complications. E. The kidney and lung represent normal tissues in which FSUs are organized in a parallel arrangement. XVIII-5) Which one of the following statements is NOT a correct characterization of radiation-induced fibrosis? A. There is a chronic overproduction of pro-inflammatory cytokines. B. There is a chronic overproduction of pro-fibrotic cytokines and growth factors. C. There are chronic increases in reactive oxygen and nitrogen species. D. Once present, radiation-induced fibrosis is fixed and untreatable. E. Complex and dynamic interactions between several cell types are involved in the development of fibrosis. Page 60 60 XVIII-6) Which of the following cytokines or growth factors is least likely to be associated with the development of radiation-induced late effects? A. TNF-α B. IL-1 C. IL-6 D. TGFβ E. VEGF XVIII-7) Which of the following statements about cytokines is FALSE? A. Each cytokine has a distinct and independent role in cell signaling. B. Cytokine signaling occurs when a ligand binds to its receptor. C. Many cytokine receptors are found both bound to cell membranes and in soluble form. D. Tyrosine kinases play a critical downstream role in the transduction of cytokine signaling. E. Despite their role in the etiology of radiation injury, many cytokines have been shown experimentally to act as radioprotectors. XVIII-8) Which of the following statements concerning stem cells is FALSE? A. In both humans and rodents, stem cells rarely maintain telomerase activity, whereas their progeny, the progenitor cells, do. B. A stem cell is an undifferentiated cell that can produce daughter cells capable of either remaining stem cells, or committing to a pathway leading to differentiation. C. Contrary to conventional thinking, it now appears that most, if not all, adult organs contain stem cells, or at least can produce stem cells in culture. D. Stem cells are said to be “pluripotent” when they can produce all the cell types of the embryo, including germ cells. E. Most tumors are thought to contain a small population of putative “cancer stem cells” capable of self-renewal and differentiation, albeit poorly- regulated. Page 61 61 XVIII-9) Which of the following statements concerning clinical applications of stem cells is FALSE? A. In the 1960s, Till and McCulloch demonstrated that mouse bone marrow contained rare cells that formed colonies in the spleens of irradiated mice, some of which contained cells that were self-renewing. B. Most tumors are thought to contain “cancer stem cells” capable of self- renewal and differentiation (although aberrant), and that these should be the ultimate targets of cancer therapy. C. There is little evidence to suggest that stem cells derived from adult tissues are sufficiently “plastic” to form anything other than cell types characteristic of the tissue from where they were derived. D. Nuclear transfer technology allows the production of embryonic stem cell lines from patients with genetically determined or influenced diseases that might thereby be “correctable”. E. One great promise of stem cell therapy is the prevention or amelioration of normal tissue complications of cancer treatment. XVIII-10) Which of the following statements regarding hierarchical tissues is FALSE? A. The time to expression of radiation-induced injury is relatively insensitive to dose. B. After a single radiation dose, the time to expression of injury depends on cellular turnover rate. C. The severity of early injury after a course of fractionated radiation therapy depends on overall treatment time. D. The time to expression of injury is a reflection primarily of cellular radiation sensitivity. E. Loss of tissue function is a result of insufficient replacement of functional cells because of damage to stem and/or progenitor cells. Page 62 62 XIX. Therapeutic Ratio XIX-1) The slope of a tumor control probability (TCP) curve is likely influenced by all of the following, EXCEPT: A. presence of tumor hypoxia B. number of clonogens in the tumor C. tumor size D. volume of normal tissue in the field E. tumor histopathological type and grade XIX-2) One would expect to achieve the largest therapeutic gain using a: A. radiation sensitizer that increases response of tumors and dose limiting normal tissue by the same factor B. radiation protector that protects the tumor but not the normal tissue C. high LET ion beam with a tumor type having a large hypoxic fraction and little reoxygenation during treatment D. radiation protector that decreases the radiation response of both the tumor and normal tissues E. fractionated course of carbon ions that yields an RBE of 3.0 for the tumor and an RBE of 5.5 for the dose limiting normal tissue XIX-3) Which one of the following statements is FALSE concerning volume effects in normal tissues? A. The radiation tolerance of many tissues depends, in part, on how the tissue is organized into functional subunits. B. In a tissue such as spinal cord, where the functional subunits are organized in series, radiation response is independent of dose for relatively small volumes, but is highly dose-dependent for large volumes. C. In organs such as the kidney and liver, where the functional subunits are organized in parallel, a greater dose can be tolerated when a small volume is irradiated compared with irradiation of the whole organ. D. In tissues like skin and mucosa, a relatively high dose can be tolerated provided the treatment volume is small, because cells from an adjacent, unirradiated area can migrate into the irradiated volume. E. An advantage of clinical approaches such as IMRT and brachytherapy is that they allow higher doses to be delivered to the tumor while reducing the dose to large volumes of normal tissue. Page 63 63 XIX-4) Combining a new chemotherapy agent with radiotherapy will only be clinically successful if the drug: A. acts synergistically with radiation B. increases the response of the tumor C. does not increase injury to the dose-limiting normal tissue D. decreases the total radiation dose needed for tumor control E. increases the therapeutic ratio XIX-5) In adults, the whole breast TD5/5 is approximately 50 Gy. For pre-adolescent girls, the TD5/5: A. increases to ~ 60 Gy B. remains unchanged C. decreases to ~ 25 Gy D. decreases to ~10 Gy E. decreases to ~ 1 Gy XIX-6) Which of the following would be expected to decrease the risk of radiation pneumonitis resulting from total body irradiation prior to bone marrow transplantation? A. administration of cyclophosphamide B. delivering the radiation in 6 fractions rather than as a single fraction C. performing an allogeneic, rather than an autologous, transplant D. infection with cytomegalovirus E. limiting the total lung dose to 50 Gy XIX-7) A tumor contains approximately 10 7 cells, 1% of which are clonogenic. The effective cell survival curve is exponential and characterized by a D10 of 5 Gy. What total dose would result in a 90% chance of tumor control if radiation therapy is not initiated until the tumor has doubled its diameter? A. 30 Gy B. 35 Gy C. 40 Gy D. 45 Gy Page 64 64 XIX-8) For fractionated radiotherapy, each successive dose to the tumor results in the death of: A. progressively fewer cells B. progressively more cells C. approximately the same number of cells D. a variable number of cells E. only S phase cells XIX-9) The shape of a tumor control probability curve for a series of identical tumors, as a function of total dose above a particular threshold, would best be described as: A. parabolic B. sigmoidal C. linear D. bell-shaped E. linear-quadratic XIX-10) The ratio of the number of cells in a 6 mm diameter solid tumor compared with a 2 mm diameter tumor will be closest to: A. 3 B. 6 C. 9 D. 18 E. 27 XIX-11) A tumor contains 10 8 clonogenic cells and has an effective D10 of 6 Gy. What dose is necessary to reduce the number of clonogens to 100? A. 6 Gy B. 12 Gy C. 24 Gy D. 36 Gy Page 65 65 XIX-12) For a particular tumor whose clonogens exhibit an effective D10 of 10 Gy, the TCD50 is 70 Gy. What would the TCD50 be if roughly 90% of the tumor mass was excised prior to the irradiation? A. 30 Gy B. 40 Gy C. 50 Gy D. 60 Gy E. 70 Gy Page 66 66 XX. Time, Dose, Fractionation XX-1) Two of your four radiotherapy machines are out of action for 10 days. Your staff is overstretched and unable to work the extra shifts necessary to treat all the patients. Which of the following options will be least likely to adversely affect treatment outcome for the patients? A. Delay initiating treatment of new patients until your machines have been repaired. B. Reduce the treatment time per day for each patient by treating only some of the designated fields each day, but compensate by boosting the dose to each field, so that the tumor dose remains constant and total treatment time remains the same. C. Introduce a break in treatment for patients who have received at least half the prescribed number of fractions, and reinitiate their treatment when the machines have been repaired. D. Reduce overall treatment time for each patient by increasing fraction size and reducing the number of fractions given. Determine the revised dose schedule through a biologically effective dose calculation. E. Reduce patient load on your remaining machines by treating your patients only 3 times per week maintaining the originally prescribed fraction size. XX-2) You propose to change your usual fractionation schedule of 2 Gy given once per day to a hyperfractionated regimen consisting of 2 fractions of 1.4 Gy each per day. What is the minimum acceptable time interval between the fractions? A. 12 hrs B. 6 hrs C. 3 hrs D. 1 hr E. 0.5 hr Page 67 67 XX-3) ARCON is best described by which of the following statements? A. ARCON (accelerated radiotherapy, carbogen, nimorazole) combines accelerated fractionated radiotherapy to counteract cellular repopulation, the hyperoxic gas carbogen (95-98% O2 + 2-5% CO2) to reduce diffusion- limited hypoxia and nimorazole, a hypoxic cell sensitizer, to reduce hypoxia-related radioresistance. B. ARCON (accelerated radiotherapy, controlled oxygen, nicotinamide) combines accelerated fractionated radiotherapy to counteract cell cycle reassortment, controlled oxygen (50% O2 via CPAP) to reduce diffusion- limited hypoxia and nicotinamide, a vasodilator, to reduce perfusion- limited hypoxia. C. ARCON (accelerated radiotherapy, controlled oxygen, nimorazole) combines accelerated fractionated radiotherapy to counteract cellular repopulation, controlled oxygen (50% O2 via CPAP) to raise hemoglobin saturation and nimorazole, a hypoxic cell sensitizer, to reduce hypoxia- related radioresistance. D. ARCON (accelerated radiotherapy, carbogen, nicotinamide) combines accelerated fractionated radiotherapy to counteract cellular repopulation, the hyperoxic gas carbogen (95-98% O2 + 2-5% CO2) to reduce diffusion- limited hypoxia and nicotinamide, a vasodilator, to reduce perfusion- limited hypoxia. E. ARCON (accelerated radiotherapy, carbogen, nitric oxide) combines accelerated fractionated radiotherapy to counteract cell cycle reassortment, the hyperoxic gas carbogen (95-98% O2 + 2-5% CO2) to reduce diffusion-limited hypoxia and nitric oxide donors (e.g. nitroglycerine) to reduce perfusion-limited hypoxia. XX-4) Which one of the following pairs of approximate α/β ratios and tissue or tumor type is INCORRECT? A. 8-10 Gy - cure/control of fibrosarcoma B. 1-2 Gy - skin erythema C. 1-3 Gy – spinal cord myelopathy D. 8-10 Gy – shrinkage of villi in the jejunum E. 3-4 Gy – lung fibrosis Page 68 68 XX-5) In a conventional fractionation pattern, a head and neck cancer is treated to a total of 70 Gy in 35 fractions, given one fraction per day, 5 days a week. What is the BED for the tumor (assuming an α/β of 10 Gy)? A. 55 Gy10 B. 60 Gy10 C. 70 Gy10 D. 80 Gy10 E. 84 Gy10 XX-6) A fractionation pattern of 70 Gy in 35 fractions, one treatment a day, 5 days a week for 7 weeks yields a BED for late effects of 117 Gy3. If it is determined that the patient would be best treated by hyperfractionation, which one of the following fractionation schemes would result in the same BED for the late effects? A. 50 Gy total in 30 fractions, 5 days a week for 6 weeks B. 60 Gy total in 30 fractions, 5 days a week for 6 weeks C. 60 Gy total in 60 fractions, twice a day, 5 days a week for 6 weeks D. 75 Gy total in 35 fractions, 5 days a week for 7 weeks E. 84 Gy total in 70 fractions, twice a day, 5 days a week for 7 weeks XX-7) The slopes of isoeffect curves relating total dose to dose per fraction for late responding normal tissues exposed to carbon ions would be expected to be: A. similar to those for late responding tissues treated with X-rays B. much steeper than those for tumors treated with X-rays C. relatively flat D. highly dependent on the dose per fraction E. calculated using an RBE of 1.5 Page 69 69 XX-8) Which of the following statements is TRUE concerning time-dose- fractionation relationships? A. Fraction size is the dominant factor in determining acute effects, while overall treatment time has little influence. B. Hypofractionation involves reduction of both the dose per fraction and overall treatment time. C. A split course treatment permits recovery of late responding normal tissues. D. Fraction size and treatment time typically both determine the response of late responding tissues. E. Incomplete repair is thought to be responsible for increased late effects associated with certain accelerated fractionation regimens. XX-9) The greatest reduction in the risk of developing a late effect following exposure to a particular dose of radiation usually can be achieved through: A. the use of a protocol employing a series of small fractions, compared to fewer, larger, dose fractions B. an accelerated protocol C. use of agents that upregulate repair enzymes D. irradiation under high oxygen pressure XX-10) The use of small daily dose fractions is considered beneficial in cancer radiotherapy for all of the following reasons, EXCEPT, it: A. allows a greater opportunity for reoxygenation. B. allows repair of sublethal damage to occur in acutely responding normal tissues. C. allows progression of tumor cells into more radiosensitive stages of the cell cycle. D. permits repair of sublethal damage in tumor cells. Page 70 70 XX-11) Concerning time, dose and fractionation, which of the following statements is TRUE? A. If treatment time is extended beyond the point when an early responding normal tissue begins to exhibit compensatory proliferation, the total dose should be decreased to avoid enhancing the severity of early reactions. B. If the interval between fractions is decreased to less than 6 hours, the incidence of late normal tissue effects may decrease due to incomplete repair. C. Prolonging overall time within the normal radiotherapy range generally has a larger sparing effect on early compared with late reactions. D. Results of clinical trials have suggested that patients with tumors exhibiting Tpot values longer than 3 days would benefit from accelerated treatment. E. If the dose is titrated to maintain the same level of tumor control, then the use of a hyperfractionated schedule will likely result in a higher incidence of late effects. XX-12) For a given total dose of X-rays, which of the following effects is most likely to be enhanced if the radiation is delivered at a low dose rate over several weeks compared to the same dose deposited within a few minutes? A. chromosomal aberrations B. cell killing C. fetal effects induced during the organogenesis portion of gestation D. mutation induction Page 71 71 XXI. Brachytherapy XXI-1) The current radioisotope of choice for temporary implants for interstitial brachytherapy is: A. iridium-192 B. cesium-137 C. cobalt-60 D. iodine-131 E. americium-241 XXI-2) When short half-life radioisotopes are used for brachytherapy, the dose rate decreases significantly during the treatment regimen. This could be addressed by modifying the treatment time to maintain a prescribed dose. However, making such a simple time correction does not deliver a biologically equivalent dose primarily, because: A. longer duration treatments allow tumor cell repopulation to occur B. longer duration treatments reduce the radiosensitizing effect of cell cycle redistribution C. longer duration treatments allow more time for tumor reoxygenation to occur D. the repair of sublethal damage is incomplete for the shorter duration treatments E. the reduction in dose-rate likely encompasses a range where biological effectiveness changes rapidly with decreasing dose rate Page 72 72 XXI-3) Which one of the following statements concerning brachytherapy is TRUE? A. The linear-quadratic model predicts that the use of high dose rate compared with low dose rate intracavitary treatments should produce a therapeutic gain for treatment of a tumor with a relatively high α/β ratio when the normal tissues in the field are characterized by low α/β values. B. For breast cancer patients treated with an iridium-192 implant, the recurrence rate should decrease if the dose rate at which a specific total dose delivered was increased. C. An advantage to the use of iodine-125 and palladium-103 for permanent interstitial implants is the relatively high energy (>100 keV) of the photons emitted. D. It may be preferable to treat a tumor with a low dose rate implant if it exhibits a relatively low α/β value. E. Assuming a constant total dose, it would be expected that treatment of head and neck cancers with an iridium-192 implant delivering radiation at a dose rate lower than 0.5 Gy/hr would result in a higher tumor control rates compared with patients whose implants delivered dose rates greater than 0.5 Gy/hr. XXI-4) Which one of the following statements concerning radiolabeled immunoglobin therapy is FALSE? A. One disadvantage associated with the use of 90 Y-labeled antibodies is that the relatively low energy (<100 keV) and resulting short range of the emitted β-particles limit the cross fire effect. B. Radiation safety is an issue with the use of 131 I-labeled compounds as this isotope emits γ-rays that may pass through the patient. C. Antibody labeling with 111 In has proved useful for imaging. D. 131 I (Bexxar/tositumomab) and 90 Y (Zevalin/ibritumomab tiuxetan) are radiolabeled mouse monoclonal anti-CD20 antibodies used for the treatment of B-cell non-Hodgkin’s lymphoma. E. Recent therapies have replaced full immunoglobulins with single chain variable region fragments. Page 73 73 XXII. Radiobiological Aspects of Alternative Dose Delivery Systems XXII-1) Treatment of tumors with densely ionizing carbon ions may be advantageous over treatments with sparsely ionizing X-rays because of all of the following, EXCEPT: A. A high dose of carbon ions can be delivered to the tumor while sparing sensitive normal tissues. B. The high LET portion of the Bragg peak can be placed within the tumor volume. C. Hypoxic tumor cells would not have a survival advantage compared with aerated cells. D. Carbon ion radiotherapy is comparable in cost to traditional treatment with X-rays. E. Carbon ions have a lower OER than X-rays. XXII-2) Which of the following reasons represents the main advantage for the use of high energy protons compared with photons in cancer radiotherapy? A. Protons exhibiting a greater variation in cell cycle radiosensitivy comoared photons. B. Protons represent a high LET form of radiation. C. Protons provide a better physical dose distribution than photons. D. Protons display a substantially reduced OER. E. There is little repair following proton irradiation relative to photons. XXII-3) What is approximately the RBE of the protons used for radiotherapy? A. 0.6 B. 1.1 C. 2.8 D. 4.7 E. 7.2 Page 74 74 XXII-4) Regarding neutron radiotherapy, which of the following statements is FALSE? A. Late responding tissue morbidity discouraged the clinical use of fast neutrons from the 1940’s until the late 1960’s. B. The use of X-rays predates the use of neutrons in the treatment of cancer. C. In a randomized RTOG trial comparing neutron radiotherapy with conventional photon radiotherapy for inoperable salivary gland tumors, the complete tumor response rates were greater for neutrons. D. The results of early animal studies, in which large daily fraction sizes were used to produce acute skin effects, resulted in an underestimation of the appropriate doses to use in clinical trials of neutrons. E. A potential clinical advantage associated with the use of fast neutrons is the low OER characteristic of this form of radiation. Page 75 75 XXIII. Chemotherapeutic Agents and Radiation Therapy XXIII-1) Which of the following is an example of an agent that primarily targets the tumor stroma? A. Ibritumomab B. Gemcitabine C. Bevacizumab D. Tirapazamine E. Cyclophosphamide XXIII-2) Potential advantages of small molecule receptor tyrosine kinase inhibitors over monoclonal antibodies for anti-angiogenic therapy include all of the following, EXCEPT, the ability to: A. use of oral dosing B. target multiple receptors with a single agent C. combine these inhibitors with chemotherapy and radiation therapy D. simultaneously target stromal cells and cancer cells XXIII-3) Concerning photodynamic therapy (PDT), which of the following statements is TRUE? A. is most effective on hypoxic cells in tumors B. requires a photosensitizer, oxygen and light C. is most efficacious in vivo when short wavelength (e.g., 400 nm) light is used D. kills cells by generating hydroxyl radicals E. employs tirapazamine as the most common photosensitizer in clinical applications XXIII-4) Tumor cells obtained from patients exhibiting clinical drug resistance frequently show one or more of the following characteristics, EXCEPT: A. increased ability to repair drug-induced DNA damage B. decreased cellular uptake or increased cellular efflux of drugs C. concomitant cross-resistance to ionizing radiation D. chromosomal evidence of gene amplification, such as the presence of “double minutes” E. overexpression of p-glycoprotein on the outer cell membrane Page 76 76 XXIII-5) Given the compelling evidence that drug resistance has a genetic basis, and that drug-resistant, “mutant” cells are already present in tumors prior to the initiation of chemotherapy, which of the following clinical strategies would best reduce the selective pressure toward the drug-resistant phenotype? A. Alternate courses of chemotherapy between two equally effective, but non-cross resistant, drugs. B. Use only non-cell cycle specific chemotherapy drugs. C. Combine chemotherapy with “resistance modifying agents” that can reverse multi-drug resistance, such as the calcium channel blocker verapamil. D. Avoid the use of chemotherapy drugs derived from natural products. E. Reserve the use of chemotherapy for large tumors rather than small ones. XXIII-6) Concurrent treatment with radiation and chemotherapy may lead to improvement in the therapeutic index for a number of solid cancer types and sites. Possible mechanisms for this include all of the following, EXCEPT: A. drug-induced inhibition of the repair of radiation damage B. drug-induced killing of radiation-resistant hypoxic cells C. drug-induced changes in inherent radiation sensitivity, e.g., sensitization (of tumors) or protection (of normal tissues) D. drug-induced stimulation of tumor cell repopulation E. radiation-induced killing of drug-resistant subpopulations of tumor cells XXIII-7) Bleomycin is more toxic to oxygenated cells than to hypoxic cells. The proposed mechanism for this is: A. the more rapid proliferation of oxygenated cells coupled with the preferential toxicity of this drug to S-phase cells B. the poor penetration of the drug through avascular tissue C. the role of molecular oxygen in the action of this drug D. increased repair of drug damage in hypoxic cells E. changes in the expression of critical target genes in hypoxic cells Page 77 77 XXIII-8) Which of the following drugs is a small molecule tyrosine kinase inhibitor that targets EML4-ALK translocations that are present in some non-small cell lung cancers? A. Sunitinib (Sutent) B. Temsirolimus (Torisel) C. Ipilimumab (Yervoy) D. Crizotinib (Xalkori) E. Vemurafenib (Zelboraf) XXIII-9) Which of the following drugs is not a bifunctional alkylating agent? A. doxorubicin B. chlorambucil C. BCNU D. melphalan E. cyclophosphamide XXIII-10) Concerning antibiotics used as antineoplastic agents, which of the following statements is FALSE? A. Dactinomycin is a specific inhibitor of RNA synthesis. B. Mitomycin C is preferentially toxic to hypoxic cells. C. These drugs are usually cell cycle phase non-specific in their action. D. The liver is especially susceptible to damage caused by doxorubicin. E. These drugs often inhibit DNA synthesis. XXIII-11) The vinca alkaloids vinblastine and vincristine sulfate are cytotoxic because they inhibit: A. protein ubiquitination B. RNA synthesis C. protein synthesis D. DNA synthesis E. microtubule polymerization Page 78 78 XXIII-12) Which one of the following statements concerning mechanisms of action of chemotherapeutic agents is FALSE? A. Ara-C is a competitive inhibitor of DNA polymerase once it is converted to ara-CTP. B. Methotrexate is a competitive inhibitor of cytidine deaminase. C. 5-fluorouracil (5-FU) is an inhibitor of thymidylate synthase. D. Capecitabine (Xeloda) is an orally-administered pro-drug of 5-FU. E. Bleomycin is a radio-mimetic drug XXIII-13) Potential strategies for gene therapy in the treatment of cancer include all of the following, EXCEPT: A. introducing a wild-type tumor suppressor gene B. inserting a toxin gene under the control of a tumor-specific promoter C. blocking the expression of an oncogene by introducing a gene encoding a corresponding antisense oligonucleotide D. enhancing the immunogenicity of a tumor by introducing genes that code for the patient’s major histocompatibility antigens E. enhancing the immunogenicity of a tumor by introducing genes encoding cytokines that stimulate immune effector cells XXIV-14) Which of the following statements correctly describes vorinostat [Zolinza or suberoylanilide hydroxamic acid (SAHA)]? A. Radical scavenger using the Chinese herb sanchi (Panax pseudoginseng). B. Tyrosine kinase inhibitor used to for treatment of GIST. C. Gadolinium chelate coated gold nanoparticle used for multimodal cancer imaging. D. Poly(ADP)-ribose polymerase inhibitor for monotherapy of triple-negative breast cancers. E. Histone deacetylase inhibitor approved for treatment of cutaneous T cell lymphoma. XXIV-15) Synthetic lethality is the term associated with: A. combining systemic chemotherapy with external beam radiotherapy B. use of synthetic cancer chemotherapeutic agents C. targeting cells with a specific DNA repair defect by inhibiting a second DNA repair pathway D. bimodal therapy, such as photodynamic (PDT) or boron neutron capture therapy (BNCT) Page 79 79 XXIV. Radiosensitizers, Radioprotectors and Bioreductive Drugs XXIV-1) If lymphocytes obtained from patients previously treated with a histone deacetylase inhibitor are examined by western blotting, which one of the following would most likely be observed? A. hyper-acetylation of histone proteins B. hypo-acetylation of histone proteins C. pancytopenia D. global loss of histone protein bands E. histone protein aggregates XXIV-2) Which of the following statements concerning misonidazole is FALSE? A. It is a 2-nitroimidazole. B. Irradiation in the presence of this drug reduces the survival of hypoxic cells without altering that of aerobic cells. C. Its dose-limiting toxicity is peripheral neuropathy. D. It is selectively toxic to hypoxic cells, but only after metabolic activation by the liver. E. It must be present in the tumor during irradiation to produce radiosensitization. XXIV-3) The effectiveness of the hypoxic cell radiosensitizer nimorazole in overcoming the resistance of hypoxic cells can be attributed to all of the following, EXCEPT the: A. ability to reach chronically hypoxic cells in tumors B. delivery of oxygen to hypoxic cells C. metabolic stability D. electron affinity XIV-4) Sulfhydryl compounds, such as the active form of amifostine, radioprotect cells by: A. increasing glutathione synthesis B. preventing ion pair formation C. inhibiting intracellular free radical formation D. scavenging free radicals E. increasing cellular oxygen Page 80 80 XXIV-5) Defects in DNA repair pathways can be exploited through a concept known as synthetic lethality. Which of the following is an example of synthetic lethality in cancer? A. Defects in homologous recombination, such as mutations in BRCA1, can be exploited by agents that inhibit nucleotide excision repair. B. Tumors with HER-2 amplification can be selectively targeted by agents that cause double strand breaks, including ionizing irradiation. C. Defects in mismatch repair, which are common in colon cancer, can be exploited by use of agents that target base excision repair. D. PARP inhibitors are selectively toxic to certain tumors due to exisiting mutations that interfere with homologous recombination. E. Tumors with EGFR amplification are sensitive to radiation due to reduced DNA repair capacity. XXIV-6) Which of the following pairs of drug and proposed mechanism of action is FALSE? A. Bromodeoxyuridine – Proliferating cell radiosensitizer B. Etanidazole - Hypoxic cell radiosensitizer C. Amifostine – Thiol-containing radioprotector D. Hydroxyurea – Mitosis-specific cytostatic agent E. Tirapazamine – Hypoxic cell cytotoxin Page 81 81 XXV. Hyperthermia XXV-1) The most likely mechanism for hyperthermia-induced cell killing is: A. direct damage to cellular RNA B. direct damage to cellular DNA C. protein denaturation D. sugar degradation E. inhibition of anabolic metabolism XXV-2) Which of the following statements best describes hyperthermia-induced cell killing and thermotolerance? A. The induction of thermotolerance is independent of the level of cell killing. B. Once induced in a “parent” cell, thermotolerance will be inherited by subsequent generations. C. The more toxic a heat exposure, the greater the degree of thermotolerance induced in surviving cells. D. Thermotolerance decays in about 10 minutes after a mild heat treatment that only produces modest cell killing. E. Thermotolerance develops secondary to mitochondrial damage. XXV-3) The greatest degree of radiosensitization occurs when: A. radiation and heat are delivered simultaneously B. heat is given first followed by radiation two hours later C. radiation is delivered first followed by heat 8 hours later D. heat and radiation are administered 24 hours apart, regarless of which is given first E. heat is administered 1 week after radiotherapy is completed XXV-4) Thermotolerance is best associated with increased: A. levels of molecular chaperones B. DNA repair capacity C. oxidative stress D. cell cycle progression E. induction of p53 Page 82 82 XXVI. Radiation Carcinogenesis XXVI-1) The absolute risk model for radiation carcinogenesis: A. predicted a drop in the leukemia incidence among Japanese A-bomb survivors B. predicts that the risk of developing a radiation-induced cancer is a multiplicative function of the natural incidence C. predicts a distinct crop of radiation-induced cancers after a latent period following irradiation D. accurately predicted that radiation-induced solid tumor incidence among Japanese A-bomb survivors would rise initially and then reach a plateau E. is the model favored by the BEIR V Committee for the calculation of radiation-induced excess cancer risk XXVI-2) Which one of the following statements concerning radiation-induced malignancies is TRUE? A. Younger women appear to be more susceptible to radiation-induced breast cancer than older women. B. The average latency period for radiation-induced leukemia is 30 years. C. Dose response curves for the induction of solid tumors show a clear threshold, with no excess cancers induced for doses below approximately 0.5 Gy. D. Most radiation-induced solid tumors appear within 5 years of irradiation. E. Children are less susceptible to radiation-induced solid tumors than adults. XXVI-3) All of the following types of cancer have shown a radiation-induced excess incidence among Japanese A-bomb survivors, EXCEPT: A. leukemia B. lung C. thyroid D. breast E. prostate Page 83 83 XXVI-4) The term “stochastic” is used to describe a radiation effect in which the: A. probability of occurrence is a function of time post-irradiation B. severity of the effect depends on the dose above a tissue/organ specific threshold C. probability of occurrence increases with dose, with no apparent threshold D. normal tissue tolerance is exceeded E. effect is limited to the tumor in a radiation field XXVI-5) The type of malignancy which exhibited a radiation-induced excess among Nova Scotia multiple fluoroscopy patients was: A. melanoma B. glioblastoma C. liver cancer D. thyroid cancer E. breast cancer XXVI-6) An accident occurs at a radioactive fuel processing plant resulting in the exposure of 5 million people to an average effective dose of 2 mSv. The expected number of excess cancer deaths in the irradiated population would be approximately: A. 1 B. 10 C. 25 D. 100 E. 500 XXVI-7) Which of the following statements concerning radiation-induced cancers is TRUE? A. For risk assessment purposes, the induction of solid tumors by radiation is assumed to follow a linear dose response. B. The most common radiation-induced tumors found among the survivors of the atomic bomb attacks on Hiroshima and Nagasaki are skin cancers. C. There does not appear to be an age dependency for radiation-induced breast cancer. D. The lifetime risk of cancer fatality resulting from a whole body radiation exposure is approximately 5x10 -6 per Sievert. E. The majority of radiation-induced cancers among the Japanese A-bomb Page 84 84 XXVI-8) Radiation-induced deterministic effects: A. are rarely produced by doses in the range used in radiotherapy B. generally have long latent periods that are independent of dose C. may be produced at any dose, however small D. vary in severity in a dose-dependent manner E. include genetic effects XXVI-9) In which of the following situations is prior radiation therapy the LEAST likely cause for the development of a second cancer? A. Second cancer within the irradiated field. B. Second cancer at a distance from the irradiated field. C. Leukemia within 5 years of the initial course of radiation therapy. D. Sarcoma 40 years after the initial course of radiation therapy. E. Head and neck cancer in a patient cured of a Pancoast lung cancer. Page 85 85 XXVII. Heritable Effects of Radiation XXVII-1) The best advice for men who have undergone pelvic radiography and are concerned about transmitting deleterious mutations to their offspring would be to: A. attempt conception as soon as possible after irradiation B. never attempt conception following this type of radiologic diagnostic test C. delay conception about 3-6 months after the procedure D. not be concerned about attempting conception, as the doses received by diagnostic procedures are not sufficient to cause mutations E. delay conception about 3 years after the irradiation XXVII-2) Concerning genetic effects induced by X-rays, which of the following statements is TRUE? A. Humans are more sensitive to radiation mutagenesis than rodents. B. A threshold dose exists below which mutations are not produced. C. The genetic doubling dose for humans is approximately 0.1 Gy. D. Radiation generally produces unique mutations that are not observed spontaneously. E. Some spontaneous mutations are caused by background radiation. Page 86 86 XXVIII. Radiation Effects in the Developing Embryo and Fetus XXVIII-1) For which of the following imaging procedures should a radiologist discuss with the patient the possibility of terminating a first trimester pregnancy due to the risk of a radiation-induced congenital malformation? A. abdominal CT B. pelvic series C. barium enema study D. intravenous pyelogram (IVP) E. none of the above XXVIII-2) Based on rodent data, the greatest risk of a congenital abnormality during gestation occurs when the embryo or fetus is exposed during: A. the first day post-conception B. the first week C. weeks one-six D. the second trimester E. the third trimester XXVIII-3) Which pair of human gestational stage and radiation-induced effects at 1 Sv is CORRECT? A. Preimplantation (0-9 days) – prenatal death B. Organogenesis (1-6 weeks) – mental retardation C. Early fetal period (6-15 weeks) - congenital anomalies D. Middle fetal period (15-25 weeks) - intrauterine death E. Late fetal period (20-40 weeks) – gross congenital malformations XXVIII-4) Epidemiologic studies of the Japanese atomic-bomb survivors irradiated in utero suggest that: A. their lifetime cancer risk was approximately 10-fold higher than the general population B. liver abnormalities were the most commonly observed congenital malformation C. radiation did not cause permanent growth retardation D. the main period of sensitivity for radiation-induced mental retardation was during the 8-15 week period of gestation E. radiation did not produce abnormalities for doses lower than approximately 2 Gy Page 87 87 XXIX. Radiation Protection XXIX-1) The average annual background radiation dose from natural sources (including radon) to the U.S. general public is: A. 3 μSv B. 100 μSv C. 3 mSv D. 10 mSv E. 0.3 Sv XXIX-2) Which of the following statements is FALSE concerning the treatment of people who arrive at the emergency room following an accident at a nuclear power plant that resulted in the release of a significant quantity of radioactive material? A. Contaminated patients should be handled using standard protective clothing and gloves. B. It is important to regulate the entry and exit from the hospital’s designated “radiation emergency area” to prevent spread of radioactive materials. C. People who are only contaminated with radionuclides but not otherwise injured can be treated in non-medical facilities. D. The primary concern for most people living within 100 miles of the accident will be an increased risk of cancer and genetic effects. E. Patients need to be decontaminated before being treated for life- threatening injuries. XXIX-3) Non-occupational radiation exposure: A. should not be permitted B. is allowed for education and training purposes C. should be limited to 10 mSv per year D. should be limited to 50 mSv per year E. should assume 1 mSv per year as the negligible individual dose Page 88 88 XXIX-4) According to NCRP recommendations, which of the following pairs of maximum permitted annual dose and population group, is correct? A. no exposure permitted - a 16 year old student performing research for a school project B. 0.5 mSv - infrequent exposure for a member of the general public C. 0.1 mSv - pregnant radiation worker D. 50 mSv - radiation worker E. 1 μSv – continuous exposure for members of the public XXIX-5) Which pair of dose type and definition is correct? A. Committed equivalent dose – average dose multiplied by a radiation weighting factor B. Equivalent dose – dose resulting from internalized radionuclides integrated over a 50 year period C. Effective dose – energy deposited per unit mass D. Collective effective dose – product of the average effective dose and the number of individuals exposed E. Genetically significant dose – average dose to the gonads XXIX-6) The genetically significant dose (GSD) for diagnostic imaging procedures will be reduced by each of the following measures, EXCEPT: A. using imaging modalities that do not employ ionizing radiation for post- menopausal women B. avoiding patients younger than 20 years of age C. using the smallest possible field size for procedures involving the lower abdomen D. collimating the beam to avoid irradiating the gonads E. using gonadal shielding for male patients Page 89 89 XXIX-7) If a terrorist detonates a radiological dispersal device (RDD) or “dirty bomb” in a crowded part of a large city, which of the following statements is TRUE? A. It is likely that most people within a 10 mile radius will die as a result of the explosion. B. Exposed individuals, regardless of any injuries they may have sustained in the blast, should be treated outside of a hospital setting due to contamination concerns. C. Individuals not killed in the initial blast will likely die within days due to contamination with radioactive materials. D. Radiation experts, including radiation oncologists, should seek media access in order to encourage all citizens living within a 100 mile radius of the detonation to obtain immediate medical attention at the closest hospital. E. If the explosion was shown to result in the release of radioactive iodine, potassium iodide should be administered as soon as possible after exposure in order to block thyroid uptake of the radionuclide. XXIX-8) The greatest single source contributing to the total average effective dose for members of the general public in the United States resulting from radiation exposure is thought to be: A. cosmic rays B. medical procedures C. nuclear power plant emissions D. radon gas E. plasma and LCD television sets Page 90 90 XXX. Molecular Techniques used in Radiation and Cancer Biology XXX-1) Which of the following methods can be used to observe nuclear foci of DNA repair-related proteins in irradiated cells? A. Comet assay B. Immunofluorescence of phosphorylated H2AX C. Single-stranded conformational polymorphism D. Induction of p53 E. Filter elution assay XXX-2) Which of the following techniques can be used to monitor gene expression? A. Electrophoretic mobility shift assay (EMSA) B. Comet assay C. cDNA microarrays D. Southern blot E. FISH XXX-3) Translation is the process in which: A. DNA is made into RNA B. protein is made from DNA C. protein is made from RNA D. DNA is made from RNA E. tRNA is made from RNA XXX-4) All of the following assays are used to assess gene expression, EXCEPT: A. Northern blotting B. Southern blotting C. Microarray D. Reverse transcriptase (RT)-PCR E. RNase protection assay XXX-5) All of the following techniques detect protein-protein interactions, EXCEPT: A. Far-western blot B. Two-hybrid screen C. Immunoprecipitation D. Electrophoretic mobility shift assay (EMSA) E. Phage display Page 91 91 XXX-6) Restriction endonucleases are NOT: A. used for DNA mapping B. enzymes that make cuts at palindromic sequences C. a type of RNA that has enzymatic activity D. found in bacteria E. used to detect restriction fragment length polymorphisms XXX-7) Which technique would you use to quantify transcript levels of a certain gene in a tumor biopsy? A. Western blot analysis B. Southern blot analysis C. Southwestern blot analysis D. Northern blot analysis E. DNA fingerprinting XXX-8) Which of the following methods CANNOT be used to identify DNA double- strand breaks? A. Determining DNA fragment size using pulsed field gel electrophoresis B. Measuring the average molecular weight of DNA using neutral sucrose gradient centrifugation C. Determining the fraction of DNA able to migrate using the neutral comet assay D. Quantifying the fraction of DNA able to elute through a filter under alkaline conditions E. Identifying the presence of microscopic foci containing phosphorylated histone H2AX (γ-H2AX). XXX-9) Which molecular technology represents the most appropriate approach to identify p53-dependent, radiation-inducible, novel transcripts in p53 wild-type versus p53-null cells? A. Comparative genomic hybridization array B. Polymorphism analysis array C. Real-time reverse transcription polymerase chain reaction D. Two-dimensional gel electrophoresis followed by DNA sequencing E. Expression array Page 92 92 XXXI. Molecular Imaging XXXI-1) Compared to the Earth’s magnetic field, how many times stronger are the magnets in an MRI machine: A. 2 B. 200 C. 20,000 D. 2,000,000 XXXI-2) Which statement comparing CT with PET scans is FALSE? A. PET uses postitron-emitting radioisotopes, whereas CT uses X-rays for imaging. B. Radioisotopes used for PET typically have a short half-life C. Both PET and CT detect changes in cell metabolism and biochemistry. D. PET and CT both pose radiation risks. E. Spatial resolution of both techniques is in the mm range. XXXI-3) Which statement comparing magnetic resonance imaging (MRI) with computed tomography (CT) is FALSE? A. MRI is best suited for imaging soft tissue. B. Bones do not generate CT image data, whereas MRI provides good details about bone structures. C. Most MRI machines can produce images in any plane without moving the patient, which is not as readily possible with a CT scanner. D. Eye cosmetics must be removed before an MRI but not before CT. E. Contrast agents can be used for both techniques. XXXI-4) Which statement about positron-emitting isotopes is TRUE? A. Positron emitting radioisotopes are created within the body along the proton path during proton radiotherapy. B. There is no shift in atomic number between the parent nucleus and its daughter product following positron emission. C. Nuclei that decay by positron emission usually have fewer protons and more neutrons than compatable with stability. D. Photons resulting from positron annihilation mainly lose their energy via photoelectric absorption in tissue. E. Methods for reconstructing images for PET have not yet been developed. Page 93 93 XXXI-5) The Hounsfield Unit Scale is used for imaging using which of the following techniques? A. PET B. MRI C. CT D. Optical imaging E. Nuclear medicine XXXI-6) Which of the following is true about why CT is used for RT treatment planning? A. The soft tissue contrast is better than MRI B. Image intensities can be used to predict dose deposition C. It can be used with “time of flight” approaches to improve image quality D. CT can be integrated with MRI to provide improved image resolution XXXI-7) Which of the following could be used to provide X-ray contrast? A. FDG B. Nanomaterials that contain gold C. Compounds containing unstable radioactive isotopes such as 11-C or 64-Cu D. Iron-containing compounds E. Fluorescent molecules XXXI-8) Which of the following radiotracers would be most useful to assess tumor cell proliferation? A. [18F]fluoro-2-deoxyglucose (FDG) B. [18F]fluorothymidine (FLT) C. [18F]fluoromisonidazole (FMISO) D. [18F]fluoroazomycin-arabinoside (FAZA) XXXI-9) Dynamic imaging by CT after delivery of a contrast agent can be used to image: A. DNA damage and quantify apoptosis B. immune cells and quantify inflammation C. vasculature and quantify perfusion D. necrosis and quantify hypoxia E. tumor cells and quantify glycolysis Page 94 94 ANSWERS, EXPLANATIONS AND REFERENCES Page 95 95 GENERAL REFERENCES Please note: Unless specific references are indicated along with the answer and explanation to a question, the material addressed in each question can be found in one or more of the following textbooks: The major textbook used in radiation biology is: Hall EJ and Giaccia AJ, Eds. Radiobiology for the Radiologist, 7th Ed. Lippincott Williams & Wilkins, Philadelphia, 2012. Additional useful textbooks include: Joiner MC, van der Kogel AJ. (eds). Basic Clinical Radiobiology, Fourth Edition, Arnold, London, 2009. Lehnert S, Biomolecular Action of Ionizing Radiation. Taylor and Francis, London, 2007. Dale R and Jones B, Ed. Radiobiological Modelling in Radiation Oncology. The British Institute of Radiology, 2007. Tannock IF, Hill RP, Bristow RG, et al., Eds. The Basic Science of Oncology, 4th Ed. McGraw-Hill, Medical Pub. Division, New York, 2005. (5 th edition available June, 2012) Mettler FA and Upton AC, Eds., Medical Effects of Ionizing Radiation, 3rd Ed. W.B. Saunders, Philadelphia, 2008. Note: Those whose primary language is Chinese may find a new Chinese language version of Basic Clinical Radiobiology to be of value: Joiner MC, van der Kogel AJ. (eds). Basic Clinical Radiobiology, Fourth Edition, Simplified Chinese Edition. (Beijing: Military Medical Science Press, ISBN 978-7-80245-435-4), 2010. Chapters on radiation and cancer biology in the major radiation oncology textbooks: Haffty B and Wilson L, Eds., Handbook of Radiation Oncology: Basic Principles and Clinical Protocols. Jones and Bartlett Publishers, Sudbury MA, 2009. Page 96 96 Halperin EC, Perez CA, Brady LW, et al., Eds., Principles and Practice of Radiation Oncology, 5 th Ed. Lippincott Williams & Wilkins, Philadelphia, 2007. Gunderson LL and Tepper JE, Eds., Clinical Radiation Oncology, Second Edition Churchill Livingstone, New York, 2007. (Third Edition to be published in 2012). Hoppe R, Phillips TL and Roach M, Eds., Leibel and Phillips Textbook of Radiation Oncology, 3rd Ed. W.B. Saunders, Philadelphia, 2010. Note: Those whose primary language is Spanish may find a new Spanish language Radiation Oncology textbook to be of value: N Urdaneta, A Vera, R E Peschel, and L H Wilson,. Radiotheripa Oncologia, Enfoque Multidisciplinario. This textbook includes chapters reviewing different aspects of different areas of clinical radiation oncology. Editorial Disinlimed, Columbia, 2009. The following represent overviews of radiation and cancer biology that are recommended: Hong WK, Bast RC, Hait WN et al, Eds., B.C. Cancer Medicine 8. Decker, Hamilton, 2010. Wilson PF and Bedford JS. Chapter 1. Radiobiologic Principles, in Leibel and Phillips Textbook of Radiation Oncology, 3rd Ed. W.B. Saunders, Philadelphia, 2010. Hammond EM, Pires IM and Giaccia AJ. Chapter 2. DNA Damage and Repair, in Leibel and Phillips Textbook of Radiation Oncology, 3rd Ed. W.B. Saunders, Philadelphia, 2010. Bentzen SM. Chapter 3. Fractionation Effects in Clinical Practice, in Leibel and Phillips Textbook of Radiation Oncology, 3rd Ed. W.B. Saunders, Philadelphia, 2010. Koch CJ, Parliament MB, Brown JM and Urtasum RC. Chapter 4. Chemical Modifiers of Radiation Response, in Leibel and Phillips Textbook of Radiation Oncology, 3rd Ed. W.B. Saunders, Philadelphia, 2010. Begg AC. Chapter 5. Prediction of Radiation Response, in Leibel and Phillips Textbook of Radiation Oncology, 3rd Ed. W.B. Saunders, Philadelphia, 2010. Chinnaiyan P, Wilson GD and Harari PM. Chapter 6. Radiotherapy and Chemotherapy, in Leibel and Phillips Textbook of Radiation Oncology, 3rd Ed. W.B. Saunders, Philadelphia, 2010. Page 97 97 Rosenstein BS. Chapter 2. The Biologic Basis of Radiotherapy: in Handbook of Radiation Oncology: Basic Principles and Clinical Protocols. Haffty B and Wilson L, Eds. Jones and Bartlett Publishers, Sudbury MA, 2009. Rosenstein BS. Chapter 3. Molecular Radiobiology: in Handbook of Radiation Oncology: Basic Principles and Clinical Protocols. Haffty B and Wilson L, Eds. Jones and Bartlett Publishers, Sudbury MA, 2009. McBride WH and Withers R. Chapter 2. Biological Basis of Radiation Therapy. pp. 76 108, in Principles and Practice of Radiation Oncology, 5 th Ed. Halperin EC, Perez CA, Brady LW, et al., Eds., Lippincott Williams & Wilkins, Philadelphia, 2007. Baumann M, Cordes N, Hasse M and Zips D. Chapter 3. Molecular Cancer and Radiation Biology. pp. 109-125, in Principles and Practice of Radiation Oncology, 5 th Ed. Halperin EC, Perez CA, Brady LW, et al., Eds., Lippincott Williams & Wilkins, Philadelphia, 2007. Jain RK and Kozak KR, Chapter 4. Molecular Pathophysiology of Tumors, pp.126-141, in Principles and Practice of Radiation Oncology, 5 th Ed. Halperin EC, Perez CA, Brady LW, et al., Eds., Lippincott Williams & Wilkins, Philadelphia, 2007. Zeman EM, Chapter 1. Biologic Basis of Radiation Oncology, pp. 1-46, in Clinical Radiation Oncology, Second Edition. Gunderson LL and Tepper JE, Eds. Churchill Livingstone, New York, 2007. Constine LS, Milano MT, Friedman D, et al. Late Effects of Cancer Treatment on Normal Tissues pp. 320-355, in Principles and Practice of Radiation Oncology, 5 th Ed. Halperin EC, Perez CA, Brady LW, et al., Eds., Lippincott Williams & Wilkins, Philadelphia, 2007. Page 98 98 I. Interaction of Radiation with Matter I-1) E The reactions of hydroxyl radicals formed from water radiolysis account for the majority of biologically-relevant damage induced in cellular constituents by radiation. I-2) C The greater the LET of the type of radiation, the more likely it is to produce clustered lesions and the more complex the lesions will be. For a given dose of low LET radiation, single strand breaks are produced in greater abundance than double strand breaks. DNA-lipid crosslinks are rarely induced by ionizing radiation. Cells irradiated in the absence of oxygen are more resistant to the killing effect of radiation. Thiol compounds are radical scavengers, thus, decreasing their intracellular concentration sensitizes cells to radiation killing. Hada M, Georgakilas AG. Formation of Clustered DNA Damage after High-LET Irradiation: A Review. J Radiat Res 49(3):203-10, 2008. PubMed I-3) D Only about a quarter to a third of the damage produced in cellular macromolecules arises through direct radiation action, i.e., production of ionizations directly in the target molecules. The majority of damage is the result of indirect action. Damage to cellular proteins and lipid peroxidation following biologically relevant doses appears to be of relatively minor importance. I-4) D The probability of the photoelectric effect increases with the atomic number of the absorber. I-5) D The energy average LET for 14 MeV neutrons is approximately 75-100 keV/µm, considerably greater than the calculated track average LET. The energy average LET better reflects the biological potency of this form of radiation. I-6) B LET is inversely proportional to the incident energy of the particle. Page 99 99 I-7) A The LET of clinically-used 150 MeV protons is only about 0.5 keV/µm. Accordingly, the biologic properties of these high energy protons are similar to those of X-rays. Groesser T, Chun S, Rydberg B. Relative biological effectiveness of high-energy ion ions for micronuleus formation at low doses. Radiat Res 168(S):675-82, 2007. PubMed I-8) C The indirect effect of radiation is the result of the interaction of radiation with cellular water molecules causing their radiolysis and resulting in the production of hydroxyl and other reactive free radical species, which then react with the target molecule. Page 100 100 II. Molecular Mechanisms of DNA Damage II-1) E Although there exists some evidence that DNA thymine dimers are produced in cells following exposure to very high X-ray doses, these DNA alterations are principally associated with exposure to ultraviolet radiation. II-2) D Clustered lesions (multiply damaged sites in close proximity), which include DNA double strand breaks, can lead to chromosome breaks. If unrejoined, they cause cell death. Other lesions listed, if distributed throughout the genome, are much less likely to cause cell killing. However, they can be components of a multiply damaged site. Georgakilas AG. Processing of DNA damage clusters in human cells: current status of knowledge. Mol Biosyst 4(1):30-5, 2008. PubMed II-3) C The yield of DNA double strand breaks for γ-rays following a dose corresponding to the D0 (1-2 Gy for most mammalian cells) is about 40 per diploid mammalian cell per Gy. The number of single-strand breaks and base damages are both about 25 times higher. II-4) B Clustered DNA lesions, which can be composed of DNA base damages, abasic sites, single strand breaks and/or double strand breaks, become progressively more complex with increasing LET of the radiation. Clustered lesions (multiply damaged sites) are thought to be particularly deleterious biologically because they are not easily or accurately repaired as genetic information in the DNA sequence is often lost. Foci of DNA repair-related proteins such as 53BP1 are used as surrogates for DNA DSBs and therefore do not specifically identify clustered lesions. Mismatch repair is not thought to be involved in repair of any type of DNA damage induced by ionizing radiation. Hada M, Georgakilas AG. Formation of Clustered DNA Damage after High-LET Irradiation: A Review. J Radiat Res, 49(3):203-10, 2008. PubMed II-5) A A DNA double strand break results from a cluster of ionizations within a single electron track (arising from a single photon interaction). Because of the very small diameter of DNA, the probability of two separate electron tracks overlapping in such a small target volume is negligible, even at high therapy doses, so a quadratic component is not seen. Page 101 101 II-6) C Some dicentric chromosme will result from a single electron track (arising from a single photon interaction) that caused two double strand breaks, one in each of two chromosomes (i.e. the linear component). Other dicentrics form from two double strand breaks caused by two separate electron tracks, from two separate photons (i.e. the quadratic component). Page 102 102 III. Molecular Mechanisms of DNA Repair III-1) D In normal cells and after DNA replication, homologous recombination involving sister chromatids is considered to be largely “error-free”. The other listed pathways have been associated with greater likelihoods of errors. The error rate would also be affected, however, by other genetic alterations within the cell such as loss of p53 (TP53). Moynahan ME, Jasin M. Mitotic homologous recombination maintains genomic stability and suppresses tumorigenesis. Nat Rev Mol Cell Biol 11(3):196-207, 2010. PubMed Branzei D, Foiani M. Maintaining genome stability at the replication fork. Nat Rev Mol Cell Biol 11(3):208-19, 2010. PubMed Negrini S, Gorgoulis VG, Halazonetis TD. Genomic instability — an evolving hallmark of cancer. Nat Rev Mol Cell Biol 11(3):220-8, 2010. PubMed Lieber MR. The mechanism of human nonhomologous DNA end joining. J Biol Chem 283(1):1-5, 2008. PubMed Powell SN, Kachnic LA. Therapeutic exploitation of tumor cell defects in homologous recombination. Anticancer Agents Med Chem. 8(4):448-60, 2008. PubMed Li X, Heyer WD. Homologous recombination in DNA repair and DNA damage tolerance. Cell Res 18(1):99-113, 2008. PubMed Page 103 103 III-2) B Mutations in either MRE11 or NBS1 result in the radiation sensitivity syndromes ataxia telangiectasia-like disorder and Nijmegen breakage syndrome, respectively. Radiation induces DNA double strand breaks and the ataxia telangectasia mutated (ATM) protein initiates signaling to activate DNA strand break repair in co-ordination with the MRE11/NBS1/RAD50 (MRN) complex. In cells defective for ATM or any member of the MRN repair factors to sites of damage decreases. Similarly, BRCA1 and BRCA2 interact with RAD51, and their inactivation reduces HR-dependent DSB repair. For example, BRCA1 is required for RAD51 nuclear focus formation. In response to DNA-damaging agents, RAD51 protein relocalizes to nuclear foci that are thought to represent sites of DNA repair by HR. Williams GJ, Lees-Miller SP, Tainer JA. Mre11-Rad50-Nbs1 conformations and the control of sensing, signaling, and effector responses at DNA double-strand breaks. DNA Repair (Amst) 9(12):1299-306, 2010. PubMed Barker CA, Powell SN.Enhancing radiotherapy through a greater understanding of homologous recombination. Semin Radiat Oncol 20(4):267-73.e3, 2010. PubMed Wang W. Emergence of a DNA-damage response network consisting of Fanconi anaemia and BRCA proteins. Nat Rev Genet 8(10):735-48, 2007. PubMed III-3) C Typically, phosphorylation of the histone H2AX occurs within minutes following irradiation. The repair of DNA double strand breaks requires the recruitment of proteins to the site of damage. ATM and DNA-PK initiate signaling that the DNA has been damaged. An early step is the phosphorylation of histone H2AX molecules near the site of the DNA strand break. Therefore, γ-H2AX is a marker for DNA double strand breaks that also has been used to monitor DNA strand break repair. γ-H2AX participates in the recruitment of repair machinery such as RAD51 and MRE11. DNA strand break repair then ensues. Sak A, Stuschke M. Use of γH2AX and other biomarkers of double-strand breaks during radiotherapy. Semin Radiat Oncol 20(4):223-31, 2010. PubMed Page 104 104 III-4) B The presence of NBS1 (the protein encoded by the gene that when mutated results in Nijmegen breakage syndrome), H2AX, SMC1 or BRCA1 nuclear foci are thought to mark the physical presence of DNA DSBs in the corresponding regions of chromatin. SOD2 (superoxide dismutase 2) is a manganese superoxide dismutase that functions to scavenge free radicals. TGFβ1 (transforming growth factor, beta 1) plays a role in regulation of cell growth and proliferation. It also induces synthesis of extracellular matrix proteins and plays a role in fibrosis. DNA polymerase alpha-primase (pol α) plays a critical role in initiating DNA replication. XPF-ERCC1 is a DNA endonuclease that incises a damaged DNA strand on the 5' side of a lesion during nucleotide excision repair and has additional roles in homologous recombination and DNA interstrand crosslink repair. Lavin MF. Ataxia-telangiectasia: from a rare disorder to a paradigm for cell signaling and cancer. Nat Rev Mol Cell Biol 9(10):759-69, 2008. PubMed III-5) D All other forms of repair listed handle damage to DNA produced by exogenous sources—radiation, UV exposure, chemical damage or endogenous oxidative damage. Only mismatch repair handles repair of mismatches that occur during the process of DNA replication. Hewish M, Lord CJ, Martin SA, et al. Mismatch repair deficient colorectal cancer in the era of personalized treatment. Nat Rev Clin Oncol 7(4):197-208, Epub 2010. PubMed Helleday T, Petermann E, Lundin C, Hodgson B, Sharma RA. DNA repair pathways as targets for cancer therapy. Nat Rev Cancer 8:193-204, 2008. PubMed III-6) E The BRCA1-BRCA2-RAD51 pathway does does not appear to play a role in translesion DNA synthesis, which permits cells to replicate DNA on a damaged template. Huen, MS, Sy SM, Chen J. BRCA1 and its toolbox for the maintenance of genome integrity, Nat Rev Mol Cell Biol 11:138-48, 2010. PubMed Gudmundsdottir K, Ashworth A. The roles of BRCA1 and BRCA2 and associated proteins in the maintenance of genomic stability. Oncogene 25(43):5864-74, 2006. PubMed Zhang J and Powell SN. The Role of the BRCA1 Tumor Suppressor in DNA Double-Strand Break Repair. Mol Cancer Res 3(10):531-9, 2005. PubMed Page 105 105 III-7) E Repair of double-strand breaks in mammalian cells occurs by both nonhomologous end joining and homologous recombination. However, because nearly all double-strand-break repair by homologous recombination occurs between sister chromatids, it takes place primarily when the region containing the double-strand break has already completed DNA replication in S phase. In contrast, NHEJ predominates in those phases of the cell cycle where a replicated sister chromatid is unavailable, i.e. G0/G1. III-8) E The core components of the mammalian nonhomologous end joining complex are the catalytic subunit of DNA protein kinase (DNA-PKcs), Ku subunits Ku70 (XRCC6) and Ku80 (XRCC5), Artemis, XRCC4 and DNA ligase IV. RAD51 is involved in homologous recombination, but has not been shown to play a role in non-homologous end-joining. Mahaney BL, Meek K, Lees-Miller SP. Repair of ionizing radiation-induced DNA double-strand breaks by non-homologous end-joining. Biochem J 417(3):639-50, 2009. PubMed III-9) B Phosphorylation of the histone variant H2AX to γ-H2AX can be accomplished by either ATM or DNA-PK (cell line and mutation specific) after the formation of radiation-induced DSBs. The attachment of γ-H2AX to the DNA near DSB forms readily visible nuclear foci. Although the specific role of γ-H2AX in DSB repair is unknown, recent reports have shown that dispersal of γ-H2AX foci correlates with repair of DSBs. It has been reported that the ability to remove γ-H2AX correlates with clonogenic survival. Bonner WM, Redon CE, Dickey JS, et al. Gamma H2AX and cancer. Nat Rev Cancer 8(12):957-67, 2008. PubMed III-10) C DNA glycosylases break the bond between the base and the deoxyribose and thus are capable of removing damaged bases. Robertson AB, Klungland A, Rognes T, et al. DNA repair in mammalian cells: Base excision repair: the long and short of it. Cell Mol Life Sci 66(6):981-93, 2009. PubMed Caldecott KW. Single-strand break repair and genetic disease. Nat Rev Genet 9(8):619-31, 2008. PubMed Page 106 106 III-11) B ATM is a protein kinase that phosphorylates proteins involved in repair of DNA double strand breaks and cell cycle checkpoint control. Among the substrates for ATM are NBS1, MRE11, H2AX, 53BP1, Chk2, FANCD2, SMC1, MDC1, BRCA1 and p53. ATM is not involved in oxygen radical scavenging, it is not a ligase, it does not sequester Bcl-2 and it is not a DNA polymerase. Bhatti S, Kozlov S, Farooqi AA, et al. ATM protein kinase: the linchpin of cellular defenses to stress. Cell Mol Life Sci 68(18):2977-3006, 2011. PubMed Derheimer FA, Kastan MB. Multiple roles of ATM in monitoring and maintaining DNA integrity. FEBS Lett 584(17):3675-81, 2010. PubMed III-12) D ICL-based chemotherapy is one of the most widely used forms of cancer treatment Deans AJ, West SC. DNA interstrand crosslink repair and cancer. Nat Rev Cancer 11(7):467-80, 2011. PubMed Page 107 107 IV. Chromosome and Chromatid Damage IV-1) E Dicentric chromosomes in peripheral blood lymphocytes can be detected following a dose as low as ~0.25 Gy. The induction of chromatid breaks by X-rays follows a linear function of dose. Chromosome-type aberrations, damage on both sister chromatids, are produced when a cell is irradiated prior to DNA synthesis. Ring chromosomes are lethal aberrations as are anaphase bridges and dicentrics. Fluorescent in situ hybridization (FISH) is able to detect stable chromosomal aberrations such as reciprocal translocations many years after irradiation. Vral A, Thierens H, Baeyens A, et al. Induction and disappearance of G2 chromatid breaks in lymphocytes after low doses of low-LET γ -rays and high-LET fast neutrons. Int J Radiat Biol 78(4):249-57, 2002. PubMed IV-2) E Dicentric chromosomes are asymmetrical aberrations, meaning there has been an unequal exchange or a loss of chromosome material. A–C are all symmetrical, chromosome-type aberrations, D is by definition not a “chromosome-type” aberration. IV-3) A Chromatid (terminal) deletions are one hit aberrations and increase linearly as dose increases. IV-4) A Reciprocal translocations are relatively stable and therefore can be detected at long times after exposure to ionizing radiation. All of the other chromosome aberrations are unstable and therefore can be detected only for a few cell generations, at most, after exposure before the irradiated cells (or their progenitors) die. Tucker JD, Cofield J, Matsumoto K, et al. Persistence of Chromosome Aberrations Following Acute Radiation: I, Paint Translocations, Dicentrics, Rings, Fragments, and Insertions. Environ Mol Mutagen 45(2-3):229-48, 2005. PubMed IV-5) D A translocation is a two-hit chromosome-type aberration. A dicentric is a two- hit chromosome-type aberration; an inversion is a two-hit chromosome-type aberration; an anaphase bridge is a two-hit chromatid-type aberration; a ring chromosome is a two-hit chromosome-type aberration. IV-6) C Chromatid aberrations are produced when cells are irradiated following DNA synthesis. Page 108 108 V. Mechanisms of Cell Death V-1) B Autophagy is a homeostatic cellular recycling mechanism that mediates removal of old or dysfunctional proteins and organelles, and is particularly important for cell survival during conditions of metabolic stress Janku F, McConkey DJ, Hong DS, et al. Autophagy as a target for anticancer therapy. Nat Rev Clin Oncol 8(9):528-39, 2011. PubMed V-2) E In the apoptotic process, endonucleases cut the DNA at precise sites in the linker region between nucleosomes, leading to the formation of fragments that are multiples of units comprising 180-200 bp. There is no cell swelling such as observed in necrosis, but rather cell shrinkage after the apoptotic process begins, followed by condensation of chromatin at the periphery of the nucleus. During the apoptotic process, the plasma membrane remains intact and later engulfs the apoptotic bodies. Apoptosis is a process of programmed cell death, which is tightly regulated. In necrosis there is usually random cleavage of DNA resulting in a mixture of different sizes of fragments. An early event occurring in apoptosis is the flipping of phosphotidyl-serine from the inner plasma membrane to the outer leaflet of the plasma membrane. Apoptosis is a physiological process involving single cells while necrosis is most often a non-physiological death involving a number of cells in an area and associated with development of inflammation. The apoptotic pathway is a highly conserved pathway, and several genes involved in apoptosis were originally discovered in the C. elegans worm. A cell will initiate the apoptotic pathway in response to a ligand such as FAS or any member of the TNF family by activating the death receptors, activating the extrinsic pathway for apoptosis. Cells that involve the mitochondrial pathway in the apoptotic response are using the so-called intrinsic apoptotic pathway. There are a number of inhibitors along the apoptotic pathway: members of the Bcl2 family that neutralize the pro-apoptotic members of this family and IAPs (Inhibitor of Apoptosis Proteins), which inhibit both activated caspase 9 and 3. Taylor RC, Cullen SP, Martin SJ. Apoptosis: controlled demolition at the cellular level. Nat Rev Mol Cell Biol 9(3):231-41, 2008. PubMed Kroemer G, Levine B. Autophagic cell death: the story of a misnomer. Nat Rev Mol Cell Biol. 9(12):1004-10, 2008. PubMed Maiuri MC, Zalckvar E, Kimchi A, et al. Self-eating and self-killing: crosstalk between autophagy and apoptosis. Nat Rev Mol Cell Biol 8(9):741-52, 2007. PubMed Page 109 109 Rupinder SK, Gurpreet AK, Manjeet S. Cell suicide and caspases. Vascul Pharmacol 46(6):383-93, 2007. PubMed Klionsky DJ. Autophagy: from phenomenology to molecular understanding in less than a decade. Nat Rev Mol Cell Biol 8(11):931-7, 2007. PubMed V-3) A There are two major types of caspases involved in the apototic process: initiator caspases and executioner caspases. The initiator caspases help propagate the apoptotic signal while the executioner caspases dismantle major components of the cell. Examples of initiator caspases are caspase 8 and 9, which can activate executioner caspases such as caspase 3, 6 and 7. None of the other enzymes mentioned are involved in apoptosis. V-4) A Mitotic catastrophe is caused by an aberrant mitosis and is associated with the formation of multinucleate, giant cells that contain uncondensed chromosomes or cells with ‘micro-nuclei’ that are lost upon division. It is not considered a ‘mode’ of cell death, but rather a ‘mechanism’ of cell death that can subsequently lead to apoptosis, necrosis, or senescence. The G2 checkpoint of the cell cycle is responsible for blocking entry into mitosis in cells that sustained DNA damage, so that the cell arrests and repairs its DNA damage before attempting to divide. However, if the G2 checkpoint is defective or if damage cannot be repaired in a reasonable period of time, the cell enters mitosis prematurely, before DNA replication is complete or DNA damage has been repaired. It then undergoes a form of mitotic catastrophe, which leads to cell death. Vakifahmetoglu H, Olsson M, Zhivotovsky B. Death through a tragedy: mitotic catastrophe. Cell Death Differ 15(7):1153-62, 2008. PubMed V-5) A Cells with elevated levels of DNA content can be found among those that have undergone mitotic catastrophe following irradiation. These cells may subsequently die by apoptosis, but could also die through necrosis or enter senescence. The other choices all represent features associated with apoptosis. Page 110 110 V-6) D It is important to distinguish between causes of cell death and modes of cell death. Mitotic catastrophe following irradiation functions as a cause of subsequent cell death which can occur through apoptosis or necrosis. Mitotic catastrophe can also lead to permanent senescence and thus loss of clonogenicity. According to studies using time-lapse video microscopy, mitotic catastrophe itself is not a mode of cell death. Some cells can continue to be metabolically active for extended periods even when they demonstrate various aspects of the mitotic catastrophe phenotype. Cells may even attempt multiple cell devisions and go through failed mitosis more than once. Eriksson D, Stigbrand T. Radiation-induced cell death mechanisms. Tumour Biol 31(4):363-72, 2010. PubMed Chu K, Teele N, Dewey MW, et al. Computerized Video Time Lapse Study of Cell Cycle Delay and Arrest, Mitotic Catastrophe, Apoptosis and Clonogenic Survival in Irradiated 14-3-3sigma and CDKN1A (p21) Knockout Cell Lines. Radiat Res 162(3): 270-86, 2004. PubMed V-7) E X-linked inhibitor of apoptosis protein (XIAP) is a member of the inhibitor of apoptosis proteins family of caspase inhibitors that selectively binds and inhibits caspases-3, -7 and -9, but not caspase-8. As such, XIAP blocks a substantial portion of the apoptosis pathway and is an attractive target for novel therapeutic agents for the treatment of malignancy. Additional proteins that regulate apoptosis, which are molecular targets for drug development, include anti-apoptotic factors Bcl2 (BCL2) and BclX (BCL2L1)L that prevent release of cytochrome c from mitochondria. Cleaved BID binds to BAX and BAK, causing the release of cytochrome c. Pro-apoptotic BCL2 family members (such as BAK, BAX, BAD and BID) promote the release of cytochrome c. Cytochrome c then interacts with apoptotic protease-activating factor 1 (APAF1), which activates caspase 9. Caspase 9 in turn activates downstream caspases such as caspase 3, leading to apoptosis. Vandenabeele P, Balluzzi L, Vanden Berghe T, et al. Molecular mechanisms of necroptosis: An ordered cellular explosion, Nat Rev Mol Cell Biol 11(10):700- 14, 2010. PubMed Cotter TG. Apoptosis and cancer: The genesis of a research field. Nat Rev Cancer 9(7):501-7, 2009. PubMed Eisenberg A, Bialik S, Simon HU, et al. Life and death partners: Apoptosis, autophagy and the cross-talk between them. Cell Death Diff 16(7):966-75, 2009. PubMed Page 111 111 Ohtani N, Mann DJ, Hara E. Cellular senescence: Its role in tumor suppression and aging. Cancer Sci 100(5):792-7, 2009. PubMed Letai AG. Diagnosing and exploiting cancer's addiction to blocks in apoptosis. Nat Rev Cancer 8(2):121-32, 2008. PubMed Youle RJ, Strasser A. The BCL-2 protein family: opposing activities that mediate cell death. Nat Rev Mol Cell Biol 9(1):47-59, 2008. PubMed Ow YP, Green DR, Hao Z, et al. Cytochrome c: functions beyond respiration. Nat Rev Mol Cell Biol 9(7):532-42, 2008. PubMed Johnstone RW, Frew AJ, Smyth MJ. The TRAIL apoptotic pathway in cancer onset, progression and therapy. Nat Rev Cancer 8(10):782-98, 2008. PubMed Riedl SJ, Salvesen GS. The apoptosome: signalling platform of cell death. Nat Rev Mol Cell Biol 8(5):405-13, 2007. PubMed V-8) E The absence of sphingomyelinase in endothelial cells has been reported to lead to a marked resistance to high-dose radiation-induced apoptotic cell death. Radiation acts directly on the plasma membrane activating acid sphingomyelinase, which generates ceramide by enzymatic hydrolysis of sphingomyelin. Ceramide then acts as a second messenger initiating an apoptotic response via the mitochondrial system. In some cells and tissues, BAX is activated downstream of ceramide, regulating commitment to the apoptotic process via release of mitochondrial cytochrome c. Kolesnick R and Fuks Z, Radiation and Ceramide-Induced Apoptosis. Oncogene 22(37):5897-906, 2003. PubMed V-9) A Increased expression and phosphorylation of p53 play a major role in the response of cells to DNA damage. This activation can lead to modified expression of genes of the BCL-2/BAX family (upregulating proapoptotic genes such as BAX and downregulating antiapoptotic genes such as Bcl2), which in certain cell types can cause mitochondrial-dependent apoptosis. Survivin is a gene that is a member of the IAP family (inhibitors of apoptosis proteins) and has been associated with increased radiation resistance in a number of cell types. Activation of EGFR may cause activation of MAP kinase pathways and resistance to apoptosis; antibodies blocking activation of EGFR such as cetuximab may induce apoptosis. PI-3K is a gene that can be activated in cells under stress, including following irradiation and can activate pathways that reduce sensitivity to apoptosis. VEGF does not play a direct role in radiation-induced apoptosis. Page 112 112 Okada H, Mak TW. Pathways of Apoptotic and Non-Apoptotic Death in Tumour Cells. Nat Rev Cancer 4(8):592-603, 2004. PubMed V-10) B Bystander effects have been observed when unirradiated cells were treated with medium taken from cell cultures previously exposed to low-dose, low- LET radiation. Bystander effects would not be demonstrated by comparing single-dose and fractionated irradiation, which would be useful to examine sublethal damage repair. The phenomenon by which cells die from excessive sensitivity to small single doses of ionizing radiation below approximately 0.2- 0.3 Gy has been termed “low-dose hyper-radiosensitivity”. Protection from a subsequent high dose exposure resulting from exposure to a low dose is termed the “adaptive response”. Adaptive responses may in certain instances be induced through a bystander effect, but could be caused by other mechanisms as well. Bystander effects have been shown after both low and high LET radiations. Effects observed generations later are considered to involve delayed mutations, not bystander effects. Blyth BJ, Sykes PJ. Radiation-Induced Bystander Effects: What Are They, and How Relevant Are They to Human Radiation Exposures? Radiat Res 176(2): 139-57, 2011. PubMed Loeb LA. Human cancers express mutator phenotypes: origin, consequences and targeting. Nat Rev Cancer 11(6);450-7, 2011. PubMed Averbeck D. Non-targeted effects as a paradigm breaking evidence. Mutat Res 687(1-2):7-12, 2010. PubMed Prise KM, O'Sullivan JM. Radiation-induced bystander signalling in cancer therapy, Nat Rev Cancer 9(5):351-60, 2009. PubMed Marples B, Wouters BG, Collis SJ, et al. Low-Dose Hyper-Radiosensitivity: A Consequence of Ineffective Cell Cycle Arrest of Radiation-Damaged G2- Phase Cells. Radiat Res 161(3):247-55, 2004. PubMed Mothersill C, Seymour C. Radiation-Induced Bystander Effects and Adaptive Responses--the Yin and Yang of Low Dose Radiobiology? Mutat Res 568(1):121-28, 2004. PubMed Mothersill C. Seymour CB. Radiation-Induced Bystander Effects--Implications for Cancer. Nat Rev Cancer 4(2):158-64, 2004. PubMed Hall EJ and Hei TK, Genomic Instability and Bystander Effects Induced by High-LET Radiation. Oncogene 22(45):7034-42, 2003. PubMed Page 113 113 Morgan WF. Is There a Common Mechanism Underlying Genomic Instability, Bystander Effects and Other Nontargeted Effects of Exposure to Ionizing Radiation? Oncogene 22(45):7094-9, 2003. PubMed Morgan WF. Non-Targeted and Delayed Effects of Exposure to Ionizing Radiation: I. Radiation-Induced Genomic Instability and Bystander Effects in Vitro. Radiat Res 159(5):567-80, 2003. PubMed V-11) A There are no immediate morphological changes that can be detected immediately following irradiation in most cells, although a characteristic of radiosensitive cell populations with a pro-apoptotic tendency is the observation of apoptosis within hours of irradiation. A sphingomyelin- dependent signaling pathway has been reported to be triggered in the membrane of endothelial cells by high-dose ionizing radiation in the absence of DNA damage, suggesting that the cell membrane may play a role in radiation-induced apoptosis in some situations. Some cycling cells will undergo up to four abortive mitotic cycles before undergoing lysis as a result of mitotic catastrophe. In contrast, those cells that enter permanent growth arrest or senescence can remain metabolically active for varying amounts of time, but do not proliferate and will eventually die by necrosis. Although many of these conditions are associated with morphological changes, the majority of cells show no morphological indication of radiation damage until they attempt to divide. V-12) D Lymphocytes in general are prone to undergoing apoptosis following irradiation, most likely through a p53-induced mitochondrial-dependent mechanism. Fibroblasts, astrocytes, hepatocytes and neurons are generally resistant to radiation-induced apoptosis. Page 114 114 VI. Cell and Tissue Survival Assays VI-1) B Cells exclude Trypan Blue if their cell membrane is intact and the active transport systems that efflux the dye are active. Cells with damaged cell membranes take up the dye and are stained blue. VI-2) D The spleen colony assay has been used to measure the radiosensitivity of bone marrow stem cells. It requires approximately 9 days and involves treatment of recipient animals with a lethal radiation dose prior to bone marrow transplantation. Till JE, McCulloch EA. A direct measurement of the radiation sensitivity of normal mouse bone marrow cells. Radiat Res 14:213-22, 2961. PubMed VI-3) A These assays require the use of radiation doses high enough, generally above about 5 Gy, such that only one clonogenic cell survives to repopulate the structural unit examined. The assays do measure cell survival, not just functional end-points, and can be used to measure the D0. They do not require explanting cells to measure colony formation because colony formation by the parenchymal cells, not vascular cells, is assayed in the irradiated tissue. Page 115 115 VII. Models of Cell Survival VII-1) D When colonies arise from units containing several cells, every clonogenic cell in the cluster would need to be killed to inhibit formation of the colony. The presence of multiple clonogenic cells within the colony-forming unit therefore would increase “n” and increase the width of the shoulder (i.e., increase Dq) on the survival curve over that which would be seen for single cells. D0 would remain unchanged. VII-2) C A single dose of 6 Gy would produce a similar level of cell killing as that produced by the total dose (10 Gy) given in two fractions minus the Dq (4 Gy). VII-3) C For X-rays, typical D0 values for aerobic cells are in the range of 1- 2 Gy, while OER values for hypoxic cells are generally 2-3. Therefore, most D0 values for hypoxic cells fall into the range of 2-6 Gy. VII-4) B The loss of reproductive integrity as measured by the ability of cells to form colonies of cells, is considered the “gold standard” for measurement of radiosensitivity for actively dividing cells. This is considered the most relevant parameter for radiotherapy because of its applicability to tumor cure. VII-5) C The D0 will equal the D37 only when there is no shoulder on the survival curve; the n will therefore equal 1. VII-6) E The Dq does not provide any information on length of S phase. VII-7) D In order to achieve a 90% probability of tumor control, there can be only a 10% probability of one clonogenic cell remaining in a tumor following radiotherapy. Therefore, since the tumor began with 10 8 clonogenic cells, the radiation must produce a surviving fraction of 10 -9 which would require a dose of 5 Gy X 9 = 45 Gy Page 116 116 VII-8) D Within the framework of the linear-quadratic model, at low doses, cell killing results from lethal lesions occurring primarily from the passage of a single ionization track that simultaneously produces breaks in two chromosomes. This can lead to formation of a dicentric. At higher doses, it becomes more likely that breaks in two chromosomes will result from separate ionization tracks. VII-9) C The resulting surviving fraction is the product of the surviving fractions from each dose. Therefore, three series of these 9 Gy regimens (5 X 1.8 Gy), which each result in a surviving fraction of 10 -3 , would reduce survival to 10 -9 (3 X 9 Gy = 27 Gy). Alternatively, calculate D10 (= 3 Gy). Nine logs of cell killing would require 9 x 3 Gy = 27 Gy. Information about the fraction size/number is not required to solve this problem. VII-10) D The Dq (which measures the shoulder width) is very small for high LET radiations. VII-11) E The number of cells already sterilized generally does not affect the sensitivity of the remaining viable cells VII-12) B A survival curve with a low α/β ratio has a relatively large β component and substantial curvature which is a reflection of killing that requires the accumulation of sublethal damage. A “breaking” survival curve in which the slope abruptly decreases is an indication of a subset of cells with differing radiosensitivities, such as might occur when there is a mix of aerated and hypoxic cells. Exponential survival curves would have a very high or undefined α/β ratio since the β value would essentially be zero or close it. In a cell population, radiation would kill a greater number of cells per Gy at low doses compared with high doses. Page 117 117 VIII. Linear Energy Transfer VIII-1) A The RBE value depends on the isosurvival/isoeffect level at which it has been calculated. The RBE values measured at high cell survival levels (that is, a “mild” endpoint) are higher than those at low cell survival levels (i.e,, more “severe” endpoint). At low doses or very low dose rates, the RBE tends towards the ratio of the α coefficients for the high LET radiation and X- rays, reaching a plateau when the dose-effect curves become indistinguishable from their tangents at the origin. This limiting value of RBE is an important factor in radiation protection. For most endpoints, the RBE for a high LET form of radiation varies with its energy. The RBE for a high LET radiation is greater for hypoxic compared with aerated cells due to the radioresistance of X-irradiated cells irradiated under hypoxic compared with aerated conditions and the relative lack of this differential response for a high LET form of radiation. Cells characterized by survival curves with small α/β ratios – indicating a large capacity for accumulating and repairing sublethal damage – tend to have large RBEs for high LET radiations, compared to cells characterized by survival curves with a larger α/β value. VIII-2) A The total dose would have the greatest impact since the other parameters listed would have relatively little effect for a high LET radiation. VIII-3) B The RBE of α-particles, which is the ratio of the X-ray dose to the dose of α- particles to produce a particular effect, is dependent on dose rate. This occurs since the X-ray dose to produce a particular biologic effect is sensitive to dose rate because of repair of damage during the irradiation, whereas the α-particle total dose to produce the effect is little changed for irradiations at different dose rates. The α-particle RBE for most effects would be greater than 2, and is generally greater for late effects than early effects. RBE is also dependent on the severity of the endpoint chosen, with RBE decreasing with increased cell kill; it is likewise greater for a fractionated compared with a single dose exposure. VIII-4) B Since the identification of (presumptive) hypoxic regions in human tumors during the 1950s, one of the main rationales for the use of high LET radiation has been to lessen the oxygen effect with respect to tumor cell killing. Particulate high LET radiations, because they are of a high molecular weight and/or are highly charged, must be accelerated to higher energies than X- rays to achieve an adequate dose to deep-seated tumors. Compared to X- rays, high LET radiations are more, not less, expensive to produce. They show an advantage in controlling some slowly growing tumors in some clinical trials and exhibit less of a cell cycle phase dependency. Page 118 118 IX. Oxygen Effect IX-1) A Since 200 MeV protons would have a similar OER to 250 kVp X-rays, tumors irradiated under clamped hypoxia would be more resistant than tumors in mice breathing air whether irradiated with protons or X-rays. The tumors in mice breathing carbogen (95% oxygen, 5% CO2) would be expected to be more sensitive – and hence more curable – than tumors in mice breathing air because more oxygen would reach the hypoxic cells in the tumor. Carbon ions are more effective at inactivating tumors (cells) than X-rays or protons and also have a relatively low OER. IX-2) B Carbon ions in the Bragg peak, where the LET would reach a peak value, would yield a very low OER. In contrast, the OER for 250 kVp X-rays is in the range of 2-3. 10 MeV neutrons would exhibit an OER of about 1.3-1.6. 160 MeV protons, even in the Bragg peak where the LET increases slightly compared with the plateau, still exhibit a relatively high OER. Carbon ions in the plateau region of the beam display a low LET and therefore a greater OER compared with the OER for cells situated in the Bragg peak. IX-3) D Bromodeoxyuridine and fluorodeoxyuridine produce radiosensitization when sufficient levels are incorporated into DNA, but the sensitization is not specific for hypoxic cells. Pimonidazole and nimorazole are both hypoxic cell sensitizers, but are only effective if present during irradiation. Amifostine scavenges free radicals and is a radioprotector. IX-4) B Oxygen can be present at a concentration well below that typically observed in arterial blood and still produce sensitization. Oxygen gradients form around isolated vessels and fall from arterial values (>50 mmHg) to almost zero at distances ranging from 100-150 µm or 10-15 cell layers. IX-5) B Oxygen’s K-value, the oxygen concentration corresponding to radiosensitivity halfway between the fully oxygenated and fully hypoxic response, occurs at a pO2 of approximately 2-10 mm Hg. Full radiosensitization occurs only if oxygen is present at the time of irradiation or added within microseconds after radiation and plateaus at pO2 levels greater than ~50 mm Hg. The OER decreases up to values of about 100 keV/μm when it reaches a minimum of 1.0 indicating that oxygen does not sensitize. Page 119 119 IX-6) D The fraction of hypoxic cells in a tumor can be estimated following exposure to a large radiation dose from the ratio of the surviving fraction of tumor cells for animals irradiated under aerated conditions divided by the surviving fraction for tumor cells derived from animals irradiated under hypoxic conditions. IX-7) E Pimonidazole, which is a 2-nitroimidazole compound whose metabolic breakdown products form adducts to biomolecules only in hypoxic cells, has been very useful in the detection and quantitation of tumor hypoxia. Cervical carcinoma patients whose tumors had pO2 levels >10 mmHg exhibited increased survival compared with patients whose tumors had lower pO2 levels. Tumor oxygen status changes as treatment progresses. Detection of patients with tumors containing hypoxic cells would be helpful for the selection of patients who would benefit from the use of a hypoxic cell radiosensitizer or a hypoxic cell cytotoxin, but not necessarily for accelerated treatment, particularly if there is little or no reoxygenation. There are several non-invasive imaging techniques (e.g., PET or SPECT) that can detect labeled nitroimidazoles and therefore regions of hypoxia in tumors. Page 120 120 X. Repair at the Cellular Level X-1) C Numerous in vitro studies have shown that the repair of sublethal damage in most cell types is complete within 2 hours. This repair time may be longer in vivo, especially for some late-responding normal tissues like spinal cord. X-2) B As dose rate is reduced from 1 Gy/min, the slope of a cell survival curve decreases (D0 increases) and the shoulder decreases (n approaches 1.0). X-3) B Melanoma cells are typically characterized by a relatively small α/β ratio for their cell survival curves after exposure to low LET radiation, such as X-rays or energetic protons, and thus display a substantial increase in cell survival for a fractionated dose compared with an acute irradiation. Cell survival curves for bone marrow stem cells exposed to low LET radiation typically exhibit a large α/β ratio and therefore relatively little sparing with fractionation. Cells, regardless of cell type, will show little or no increase in surviving fraction with a split dose when irradiated with high LET alpha particles or carbon ions. X-4) D The inverse dose rate effect is manifest as an increase in cell killing at low dose rates caused by cell cycle redistribution and arrest in the radiosensitive G2 phase of the cell cycle. It is not the result of bystander effects (occurrence of biological responses in unirradiated cells nearby or sharing medium with irradiated cells) or potentially lethal damage repair (PLDR). The inverse dose rate effect would not be expected to occur in non-proliferating cells, since it is based on cells moving through the cell cycle and being blocked in the radiosensitive G2 phase. X-5) C Evidence for a role of potentially lethal damage (PLD) occurs when modification of the post-irradiation environmental conditions results in a change in biologic outcome. If cell survival increases, this is referred to as repair of PLD whereas if survival decreases it is interpreted as expression of PLD. Page 121 121 X-6) D In most cases, a low dose rate irradiation results in less toxicity than a high dose rate exposure since sublethal damage can be repaired during the course of the irradiation. The number of free radicals formed depends mostly on total dose rather than dose rate. Although some cell division may take place during an irradiation at 1 Gy/hr, this will be minimal because of the G2 block. The principal reason that irradiation at a low dose rate is less lethal, is due to sublethal damage repair. The extent of free radical scavenging and chemical restitution are relatively unaffected by the differences in dose rate indicated. X-7) C The kinetics of the increase in survival observed with increasing time between fractions is similar to the kinetics of DNA double strand break repair. The less time between fractions, the less sublethal damage repair (SLDR) is possible. SLDR is of diminished importance for exposure to high LET compared with low LET radiations since a relatively small percentage of cells is killed due to accumulated sublethal damage. SLDR occurs both in vivo and in vitro. A change in biologic effect due to a modification of post-irradiation environmental conditions is defined operationally as either repair or expression of potentially lethal damage (PLD) rather than SLDR. X-8) E Kidney, lung, spinal cord and breast tissue are all late responding normal tissues with relatively low α/β ratios. As such, these tissues are more likely to be “spared” by lowering the dose rate or using hyperfractionation. In contrast, early responding bone marrow has a nearly exponential survival curve (high α/β) and undergoes little sparing upon lowering of the dose rate or fractionating the dose. Page 122 122 XI. Solid Tumor Assay Systems XI-1) D Of the endpoints listed, tumor growth delay provides the best metric for measuring the effects of the treatment and for comparing different treatments. Tumor dose for cure of 50% of the tumors (TCD50) would be expected to have the most relevance to the clinic. Begg AC. Analysis of Growth Delay Data: Potential Pitfalls. Br J Cancer 4:93-7, 1980. PubMed XI-2) E In the lung colony assay, the lung tumors derived from intravenously injected tumor cells are counted. This clonogenic (colony formation) assay has been used to measure cell survival after treatment for cells suspended from solid tumors of several different types. When injected intravenously, leukemia cells usually home to spleen and their survival can be measured using a spleen colony assay. XI-3) D The surviving fraction is equal to the TD50-control/ TD50-irradiated = 2/8 = 0.25 XI-4) E C3H mice that carry the mouse mammary tumor virus (MMTV) and pass it to their offspring in their milk have a high incidence of mammary carcinomas and were once widely used to study the development of breast cancers. They have normal immune systems and would reject xenografts unless heavily immunosuppressed by treatment with radiation and/or drugs. Nude mice and rats carry spontaneous mutations causing thymic involution that leads to defects in cell-mediated immunity (the same mutation leads to lack of fur, which is the reason for the name.) The immunologic defects in nude mice and nude rats are sufficiently severe that xenografted tumors will grow, although in some strains of nude mice, residual immune responses can complicate experiments. SCID mice (C.B.17) carry a mutation that leads to defective T- and B-cell immunity and also to defects in V(D)J recombination and DNA repair. Xenografted tumors grow well in SCID mice; however, the DNA repair deficit in the hosts causes the mice to be unusually sensitive to radiation and some drugs, complicating some experiments. The hamster cheek pouch is an immunologically privileged site. Xenografts implanted into this site will grow and can be used in experiments. Page 123 123 XII. Tumor Microenvironment XII-1) A Hypoxic cells are typically more resistant to low LET ionizing radiation and at least some chemotherapy agents and therefore may be more likely to survive treatment. However, even if a tumor contained cells of mixed intrinsic radiosensitivity, hypoxia would not exert greater toxicity against the radiosensitive cells. Hypoxia facilitates genomic instability, enhances the survival of cells with diminished apoptotic potential in solid tumors and inhibits cell proliferation. Laboratory studies and clinical trials suggest increased metastatic potential in hypoxic cell populations. Wilson WR, Hay MP. Targeting hypoxia in cancer therapy. Nat Rev Cancer 11(6):393-410, 2011. PubMed XII-2) B Reoxygenation of hypoxic cells within a tumor that survive a dose of radiation are known to reoxygenate and become more sensitive to a subsequent dose. Reoxygenation may occur through loss and removal of oxygenated cells, through reduction in oxygen metabolism by irradiated cells or by transient changes in blood flow. Thus, both chronic and acutely hypoxic cells can change oxygenation status between fractions. XII-3) E Experience with various methodologies to measure tumor hypoxia have revealed extensive heterogeneity in the levels amongst different patients. With oxygen electrode needle measurements, this variation ranges from 0% of readings below 5 mm Hg to 100% of readings below 5 mm Hg. Immunohistochemical detection of hypoxia has demonstated that oxygenation through the tumor is highly spatially heterogeneous as might be expected based on its association with tumor vasculature. Oxygenation levels in tumors vary from complete anoxia to normal levels, and can vary significantly over time due to changes in blood flow. XII-4) A Existing imaging modalities such as CT, MRI, PET, MRS and ultrasound are already non- or minimally-invasive. Page 124 124 XII-5) B One potential advantage of stroma-directed cancer therapies is that stromal cells are not as genetically unstable as cancer cells and are therefore less likely to develop drug resistance. The interaction of cancer cells and tumor stroma plays a critical role in several steps of carcinogenesis. Cancer cell- tumor stromal interactions lead to a constantly evolving microenvironment commonly resulting in cancer cell drug resistance. Cancer cells alter their adjacent stroma through the production of a range of growth factors and proteases to form a permissive and supportive environment for tumor progression. The resultant tumor stroma is described as “reactive.” Reactive stromal cells, in turn, can promote tumor progression through the production of growth factors, pro-migratory extracellular-matrix components, and proteases. Increasing evidence suggests tumor stroma may play a direct role in tumorigenesis by acting as a mutagen by producing oncogenic signals. Hiscox S, Barrett-Lee P, Nicholson RI. Therapeutic targeting of tumor-stroma interactions. Expert Opin Ther Targets 15(5):609-21, 2011. PubMed XII-6) A [18F]FDG is commonly used for cancer detection since F-18 is a positron emitter and therefore can be detected using PET. FDG, as a glucose analog, is taken up by high-glucose-using cells such as cancer cells, where phosphorylation prevents the glucose from being released. As a result, the distribution of 18 F-FDG is a good reflection of glucose uptake and metabolism by cancer cells in the body. Although glucose uptake can be stimulated in hypoxic cells through the HIF transcription factor, many other cellular pathways influence glucose metabolism as well. As a consequence, FDG uptake and regions of hypoxia are not correlated across patients. Pimonidazole is a 2-nitroimadazole analog of misonidazole that is reductively activated and bound to cellular macromolecules in the presence of low oxygen levels. It is detected by staining tumor biopsies or sections with a labeled monoclonal antibody. FAZA and FMISO are nitroimidazole compounds that undergo bioreductive metabolism under conditions of hypoxia producing free radical metabolites that are further reduced and bound to cell constituents under hypoxic conditions. FAZA and FMISO are labeled with 18 F for detection using PET. CU-ATSM (non-nitroimidazole) contains the positron emitter 64 Cu. Reduction of Cu(II)–ATSM takes place in both normoxic and hypoxic cells resulting in unstable Cu(I)–ATSM which slowly dissociates. Cu(I)-ATSM completely dissociates in hypoxic cells and becomes irreversibly trapped, but in the presence of oxygen (normoxic cells) the Cu(I)–ATSM is reoxidized to Cu(II)–ATSM and diffuses back out of the cells. Imam SK. Review of positron emission tomography tracers for imaging of tumor hypoxia. Cancer Biother Radiopharm 25(3):365-74. PubMed Page 125 125 Lapi SE, Voller TF, Welch MJ. PET imaging of hypoxia. PET Clin 4(1):39-47, 2009. PubMed Brindle K. New approaches for imaging tumour responses to treatment. Nat Rev Cancer 8(2):94-107, 2008. PubMed XII-7) B Because of its ability to correct anemia and improve the quality of life of cancer patients by reducing fatigue, erythropoietin (EPO) has been prescribed to cancer patients. Although EPO significantly reduces the need for red blood cell transfusions in cancer patients, some clinical studies have reported decreased survival and disease control following EPO treatment. In addition, the expression of EPO receptors on tumor cells may influence tumor growth by EPO and EPO receptor expression on endothelial cells may cause EPO stimulation of tumor angiogenesis. Thus, the impact of anemia correction on prognosis remains controversial, although anemia has been hypothesized to lead to tumor hypoxia, angiogenesis and resistance to chemotherapy and radiotherapy. Several studies have indicated that tumor oxygenation and activation of hypoxic signaling pathways (e.g. HIF-1α) are independent of hemoglobin levels. In both the German/Swiss head and neck cancer trial and the BEST trial evaluating potential benefits of exogenous erythropoietin, treatment raised hemoglobin levels but had deleterious effects on survival. In general, EPO has not been used in radiation victims, but CSFs have. Dicato M, Plawny L. Erythropoietin in cancer patients: pros and cons. Curr Opin Oncol 22(4):307-11, 2010. PubMed Tóvári J, Pirker R, Tímár J, et al. Erythropoietin in cancer: an update. Curr Mol Med 8(6):481-91, 2008. PubMed Sytkowski AJ. Does erythropoietin have a dark side? Epo signaling and cancer cells. Sci STKE 2007 (395):pe38, 2007. PubMed Page 126 126 XII-8) A Tumor vasculature is functionally abnormal and differs from normal tissue vasculature in several critical ways. Tumor blood vessels are hyperpermeable, tortuous, dilated, saccular, and have a haphazard pattern of interconnection. In addition, they are poorly and/or abnormally invested with supporting pericytes. The basement membranes found in tumor vascular beds vary from abnormally thick to entirely absent. These structural abnormalities contribute to spatial and temporal heterogeneity in tumor blood flow. Furthermore, compressive forces generated by proliferating cancer cells collapse intratumoral blood and lymphatic vessels impairing blood and lymph flow. Taken together, these vascular abnormalities yield a microenvironment characterized by elevated interstitial fluid pressure, hypoxia and acidosis, all of which can compromise the efficacy of cancer therapies. These differences can profoundly limit the ability to deliver adequate concentrations of oxygen and chemotherapeutics to the entire tumor. Goel S, Duda DG, Xu L, et al. Normalization of the vasculature for treatment of cancer and other diseases. Physiol Rev 91(3):1071-121, 2011. PubMed XII-9) E Hypoxia-inducible factor-1 (HIF-1) is a heterodimer that acts as a key regulator of several oxygen-responsive proteins, including erythropoietin and vascular endothelial growth factor. HIF-1 was first identified as a DNA- binding protein that mediated the up-regulation of the erythropoietin gene under hypoxic stress. Subsequent studies have implicated HIF-1 in the regulation of a broad range of oxygen responsive genes including VEGF, VEGF receptors, angiopoietins, nitric oxide synthase, fibroblast growth factors and platelet-derived growth factor. Under aerobic conditions, HIF-1α is hydroxylated by HIF prolyl hydroxylases. Hydroxylation at two prolyl residues targets HIF-1α to the von Hippel-Lindau E3 ubiquitin ligase resulting in HIF-1α ubiquitination and subsequent proteosomal degradation, thereby limiting upregulation of target genes. Because the hydroxylation catalyzed by prolyl hydroxylases requires molecular oxygen, HIF escapes inactivation under hypoxic conditions. Dewhirst MW. Relationships between cycling hypoxia, HIF-1, angiogenesis and oxidative stress. Radiat Res 172(6):653-65, 2009. PubMed Cao Y. Off-tumor target –beneficial site for antiangiogenic cancer therapy? Nat Rev Clin Oncol 7(10):604-9, 2010. PubMed Fandrey J, Gassmann M. Oxygen sensing and the activation of the hypoxia inducible factor 1 (HIF-1)--invited article. Adv Exp Med Biol 648:197-206, 2009. PubMed Page 127 127 Kaelin WG Jr. The von Hippel-Lindau tumour suppressor protein: O2 sensing and cancer. Nat Rev Cancer 8(11):865-73, 2008. PubMed Bertout JA, Patel SA, Simon MC. The impact of O2 availability on human cancer. Nat Rev Cancer 8(12):967-75, 2008. PubMed Bristow RG, Hill RP. Hypoxia and metabolism. Hypoxia, DNA repair and genetic instability. Nat Rev Cancer 8(3):180-92, 2008. PubMed Dewhirst MW, Cao Y, Moeller B. Cycling hypoxia and free radicals regulate angiogenesis and radiotherapy response. Nat Rev Cancer 8(6):425-37, 2008. PubMed XII-10) A Hypoxia-inducible factors control genes that encode products that control tumor invasiveness and metastasis. De Bock K, Mazzone M, Carmeliet P. Antiangiogenic therapy, hypoxia, and metastasis: risky liaisons, or not? Nat Rev Clin Oncol 8(7):393-404, 2011. PubMed Ebos JM, Kerbel RS. Antiangiogenic therapy: impact on invasion, disease progression, and metastasis. Nat Rev Clin Oncol 8(4):210-21, 2011. PubMed Ellis LM, Hicklin DJ. VEGF-targeted therapy: mechanisms of anti-tumour activity.Nat Rev Cancer 8(8):579-91, 2008. PubMed Jain RK. Lessons from multidisciplinary translational trials on anti-angiogenic therapy of cancer. Nat Rev Cancer 8(4):309-16, 2008. PubMed XII-11) A HIFs bind to DNA as heterodimers composed of an oxygen-sensitive alpha- subunit and a constitutively expressed beta-subunit, also known as the ARNT (aryl hydrocarbon receptor nuclear translocator). XII-12) A The presence of intratumoural hypoxia is a negative prognostic indicator since it has been associated with increased local failure following radiotherapy and increased distant metastatic spread. Bristow RG, Hill RP. Hypoxia and metabolism. Hypoxia, DNA repair and genetic instability. Nat Rev Cancer 8(3):180-92, 2008. PubMed Page 128 128 XIII. Cell and Tissue Kinetics XIII-1) C Following ionizing radiation exposure, virtually all proliferating cells (independent of p53 (TP53) status) will show a radiation dose-dependent arrest in the G2 phase. Typically, cells with functional wild-type p53 will also arrest in the G1 phase of the cell cycle, and some cells may also arrest in S phase after radiation. XIII-2) B In response to ionizing radiation-induced DNA damages, principally DNA DSBs, ATM undergoes a conformational change and is then autophosphorylated. The activated ATM kinase then phosphorylates the p53 transcription factor which then binds to the promoter of p21 causing this inhibitor of cyclin-dependent kinases to be up-regulated. Lee JH, Paull TT. Activation and regulation of ATM kinase activity in response to DNA double-strand breaks. Oncogene 26(56):7741-48, 2007. PubMed Lavin MF, Kozlov S. ATM activation and DNA damage response. Cell Cycle, 6(8):931-42, 2007. PubMed Kastan MB and Bartek J. Cell-Cycle Checkpoints and Cancer. Nature 432(7015):316-23, 2004. PubMed XIII-3) D Cells are generally most resistant in the latter part of S phase. XIII-4) B Most proliferating human tumor cells have a doubling time of 1-5 days. The volume doubling time of the tumor as a whole is generally much longer because not all cells in the tumor are proliferating and there is often substantial cell loss XIII-5) D Since the growth fraction = 1.0, then LI = λTs/Tc or (LI)(Tc)/λ = Ts = (0.2)(3 days)(24 hours/day)/0.7 = 20.6 hours. XIII-6) C In a percent labeled mitosis experiment, the time between the point at which 50% of the mitotic cells are labeled and the time after the peak at which the percentage falls back to 50% is good approximation of the length of S phase for those cells. Page 129 129 XIII-7) C The cell cycle time is 20 hours. Therefore, the fraction of the proliferating cells in S phase should be 5/20 or 0.25, but this has to be multiplied by 0.5 to take into account the growth fraction giving 0.125. Thus, of the 2 x 10 7 cells, it would be anticipated that (2 x 10 7 )(0.125) = 2.5 x 10 6 cells would be in S phase. XIII-8) C Ts can be estimated from the relative movement, which is derived from the fraction of labeled cells that are present in G2 at the time of biopsy and the time from infusion of the label. TS and the LI are necessary to calculate the Tpot. The biopsy is taken typically 4-8 hours after administration of the label. Unfortunately, clinical studies using this approach were not able to validate Tpot as a helpful predictor of which patients might benefit from accelerated radiotherapy. XIII-9) A CDK1 is associated with progression of cells from G2 into M. Schwartz GK, Shah MA. Targeting the Cell Cycle: A New Approach to Cancer Therapy. J Clin Oncol 23(36):9408-21, 2005. PubMed XIII-10) B G0 cells are G1 cells that have ceased (temporarily or permanently) active progression through the cell cycle and thus have a similar DNA content to G1 cells. Page 130 130 XIV. Molecular Signaling XIV-1) B RAS does not appear to be related to ATM activation. Overexpression or activation of receptors such as the epidermal growth factor (EGF) receptor increases RAS signaling. Moreover, activating mutations in the RAS pathway are found in many forms of cancer. RAS signaling activates both the mitogen-activated protein kinase (MAPK) pathway and the phosphoinositide 3 kinase (PI3K) pathway. Both of these signaling pathways increase cell viability, therefore, radiation resistance is influenced by RAS activation. RAS signaling involves farnesyltransferase; inhibitors of farnesyltransferase therefore diminish RAS-induced radiation resistance. Karnoub AE, Weinberg RA. Ras oncogenes: split personalities. Nat Rev Mol Cell Biol 9(7):517-31, 2008. PubMed Schubbert S, Shannon K, Bollag G. Hyperactive Ras in developmental disorders and cancer. Nat Rev Cancer 7(4):295-308, 2007. PubMed Sousa SF, Fernandes PA, Ramos MJ. Farnesyltransferase inhibitors: a detailed chemical view on an elusive biological problem. Curr Med Chem 15(15):1478-92, 2008. PubMed XIV-2) A ERBB2/HER-2 appears to be overexpressed in roughly 20-30% of metastatic breast cancers. Receptor tyrosine kinases, like those in the ERBB family, activate signal transduction through the PI3K/AKT signaling pathway to enhance cancer cell survival. Antagonists of the ERBB family of receptor tyrosine kinases are therefore potential radiation sensitizers. Mutations in the EGF receptor, another member of the ERBB family, can be found in malignant gliomas and lung cancer. EGF receptor kinase inhibition can be achieved using antibodies or small molecule kinase inhibitors. Trastuzumab (Herceptin) is an anti-c-ERBB2 monoclonal antibody and has become a standard component of chemotherapy for c-ERBB2-positive advanced breast carcinoma. Marmor MD, Skaria KB and Yarden Y. Signal Transduction and Oncogenesis by ERBB/HER Receptors. Int J Radiat Oncol Biol Phys 58(3):903-13, 2004. PubMed Valerie K, Yacoub A, Hagan MP, et al. Radiation-Induced Cell Signaling: Inside-Out and Outside-In. Mol Cancer Ther 6(3):789-801, 2007. PubMed Page 131 131 XIV-3) C The mammalian target of rapamycin (mTOR) is a kinase that functions as a key regulator of cellular metabolism, protein synthesis, and autophagy. mTOR activity influences cell survival and death pathways and is often activated in cancer due to mutations in upstream signaling pathways. Drugs such as rapamycin are promising cancer drugs as they suppress cell growth and surivival. mTOR has at least two main downstream consequences. The first is to stimulate protein synthesis and cell growth. The second is to suppress autophagy. The PI3K–AKT cell viability pathway activates mTOR. Rapamycin analogues that inhibit mTOR can be radiation sensitizing in some cell types and can also slow the growth of tumors in animal models without displaying significant toxicity. Although mTOR is a member of the evolutionarily conserved family of phosphoinositol kinase-related kinases (PIKK) that also includes DNA-PK, ATM and ATR, it functions primarily as a serine/threonine protein kinase. Recent literature points to an interaction between the pathways for mTOR and the ubiquitin-proteasome system in the regulation of cell growth and survival. Zoncu R, Efeyan A, Sabatini DM. mTOR: from growth signal integration to cancer, diabetes and ageing. Nat Rev Mol Cell Biol 12(1):21-35, 2010. PubMed Wouters BG, Koritzinsky M. Hypoxia signalling through mTOR and the unfolded protein response in cancer. Nat Rev Cancer 8(11):851-64, 2008. PubMed Petroulakis E, Mamane Y, Le Bacquer O, et al. mTOR Signaling: Implications for Cancer and Anticancer Therapy. Br J Cancer 94(2):195-9, 2006. PubMed XIV-4) C p53 (TP53) plays an important role in radiation-induced apoptosis through interactions with proteins such as BAX and PUMA. All the other proposed activities of p53 are false. Murray Zmijewski F, Slee EA, Lu X. A complex barcode underlies the heterogeneous response of p53 to stress. Nat Rev Mol Cell Biol 9(9):702-12, 2008. PubMed Vazquez A, Bond EE, Levine AJ, et al. The genetics of the p53 pathway, apoptosis and cancer therapy. Nat Rev Drug Discov 7(12):979-87, 2008. PubMed Sengupta S and Harris CC. p53: Traffic Cop at the Crossroads of DNA Repair and Recombination. Nat Rev Mol Cell Biol 6(1):44-55, 2005. PubMed Chipuk JE, Green DR. Dissecting p53-dependent apoptosis. Cell Death Differ 13(6):994-1002, 2006. PubMed Page 132 132 XIV-5) A MDM2 negatively regulates p53 by inhibiting p53 transcriptional activity and promoting its ubiquitin-mediated proteasome degradation. The MDM2 ring finger domain has been shown to possess E3 ligase activity and to be a necessary domain for targeting p53 degradation. Manfredi JJ. The Mdm2-p53 relationship evolves: Mdm2 swings both ways as an oncogene and a tumor suppressor. Genes Dev 24(15):1580-9. PubMed XIV-6) C Bortezomib (PS-341; Velcade) is a proteasome inhibitor. Celecoxib (Celebrex) is a COX-2 inhibitor. Rituxan is a humanized monoclonal antibody to the CD20 on B cell lymphomas. Taxol is a mitotic spindle poison. Celecoxib is a cyclooxygenase-2 (COX-2) inhibitor. Erbitux (cetuximab; IMC- C225) is a monoclonal antibody inhibitor of EGFR. Martini M, Vecchione L, Siena S, et al.Targeted Therapies: how personal should we go? Nat Rev Clin Oncol Epub ahead of print 2011. PubMed XIV-7) C Farnesyltransferase (FTase) inhibitors (FTIs) were originally developed to target GTPases such as the RAS proteins. It was hoped that FTIs would act as anticancer agents and, possibly, radiation sensitizers of cells that overexpress RAS. However, recent studies have shown that FTIs may exert their actions in a RAS-independent, and possible even an FTase- independent, fashion. Berndt N, Hamilton AD, Sebti SM. Targeting protein prenylation for cancer therapy. Nat Rev Cancer 11(11):775-91, 2011. PubMed Sousa SF, Fernandes PA, Ramos MJ. Farnesyltransferase inhibitors: a detailed chemical view on an elusive biological problem. Curr Med Chem 15(15):1478-92, 2008. PubMed XIV-8) A Of the genes listed, the only one that has been used for the purpose of creating a radiation-inducible chimeric gene used is EGR1 (early growth response-1). Laboratory and pre-clinical studies have led, for example, to development of a replication-deficient adeno-vector carrying a transgene encoding human tumor necrosis factor alpha (TNF-α) under control of the radiation-inducible promoter from EGR-1 that is now entering clinical trials. Weichselbaum RR, Kufe D. Translation of the radio- and chemo-inducible TNFerade vector to the treatment of human cancers. Cancer Gene Ther 16(8):609-19, 2009. PubMed Page 133 133 XIV-9) B Iressa (Gefitinib) is a small molecule inhibitor of the epidermal growth factor receptor (EGFR). Gleevec is an inhibitor of not only the ABL tyrosine kinase, but also the tyrosine kinase activity of KIT and PDGFR. Wortmanin is a PI3K inhibitor. Gemcitabine is a nucleoside analog that causes S-phase-specific cytotoxicity. Herceptin (Trastuzumab) is a monoclonal antibody to the HER2/NEU growth factor receptor. Pao W, Chmielecki J. Rational, biologically based treatment of EGFR-mutant non- small-cell lung cancer. Nat Rev Cancer 10(11):760-74, 2010. PubMed XIV-10) B Transfection is the direct transfer of DNA molecules into a cell and does not represent a signaling pathway. Extracellular signaling pathways begin following a trigger, frequently a cytokine or growth factor, and culminate in the cellular response, e.g. growth, differentiation, etc. The extracellular signal (ligand) binds to the membrane-spanning receptor, inducing autophosphorylation of the intracellular domains of tyrosine residues. The receptor tyrosine kinases then recruit cytoplasmic signal molecules, usually through phosphorylation-dephosphorylation interactions. The signaling activity is tightly regulated; one means of regulating receptor activity is through internalization and degradation, linked through the ubiquitin pathway. The overall process for the transmission of the signal from cell surface to the nucleus is known as signal transduction. McBride WH, Chiang CS, Olson JL, et al. A Sense of Danger from Radiation. Radiat Res 162(1):1-19, 2004. PubMed Dent P, Yacoub A, Contessa J, et al. Stress and Radiation-Induced Activation of Multiple Intracellular Signaling Pathways. Radiat Res 159(3):283-300, 2003. PubMed XIV-11) B Mutational activation of one of the three RAS proteins occurs in roughly one- third of human cancers. Vigil D, Cherfils J, Rossman KL, Der CJ. Ras superfamily GEFs and GAPs: Validated and tractable targets for cancer therapy? Nat Rev Cancer 10(12):842-57, 2010. PubMed Page 134 134 XIV-12) D ERBB-directed therapeutics have demonstrated clinical efficacy; however, the antitumour effects are often not as strong as predicted from preclinical studies. There are likely to be various reasons why this is so, an important one being that other tumor-cell alterations influence the tumour response to ERBB-targetedinhibitors. Therefore, rational drug-combination strategies have great potential to combat the complexity of tumor biology. Hynes NE, Lane HA.ERBB Receptors and Cancer: The Complexity of Targeted Inhibitors. Nat Rev Cancer 5(5):341-54, 2005. PubMed Page 135 135 XV. Cancer XV-1) A Rearrangements involving the RET gene are common in radiation-associated papillary thyroid cancer (PTC). The RET/PTC1 type of rearrangement is an inversion of chromosome 10 mediated by illegitimate recombination between the RET and the H4 genes, which are 30 megabases apart. Wells SA Jr, Santoro M. Targeting the RET pathway in thyroid cancer. Clin Cancer Res 15(23):7119-23, 2009. PubMed XV-2) C Many cancers have mutations in a variety of oncogenes and tumor suppressor genes. Experimental studies have suggested that some of these mutations may result in tumor cell radioresistance. Data for mutated RAS appears to be the most consistently associated with increased radioresistance. Mutations in the other genes listed generally have either no impact or increase radiosensitivity. Gupta AK, Bakanauskas VJ, Cerniglia GJ, et al. The Ras radiation resistance pathway. Cancer Res 61(10):4278-82, 2001. PubMed Huang S, Benavente S, Armstrong EA, et al. p53 Modulates Acquired Resistance to EGFR Inhibitors and Radiation. Cancer Res 71(22):7071-9, 2011. PubMed Pylayeva-Gupta Y, Grabocka E, Bar-Sagi D. RAS oncogenes: weaving a tumorigenic web. Nat Rev Cancer 11(11):761-74, 2011. PubMed Fackenthal JD, Olopade OI. Breast cancer risk associated with BRCA1 and BRCA2 in diverse populations. Nat Rev Cancer 7(12):937-48, 2007. PubMed XV-3) C Epigenetics refers to stable changes in gene expression NOT associated with changes in the DNA coding sequence; there are both transcriptional and post-transcriptional mechanisms for the epigenetic regulation of genes. At present, two main molecular mechanisms of epigenetic regulation are present mammalian cells: DNA methylation of cytosine (primarily CpG), and histone post-translational modifications by methylation, acetylation and ubiquitination. Both hematological and solid tumors show substantial changes in epigenetic markers, including hypermethylation of the promoter regions of many genes, hypomethylation of some repetitive sequence motifs and centromeric DNA and aberrant histone modification due to presumptive defects in the corresponding histone acetylases and/or (de)methylases. DNA hypermethylation silences genes at the transcriptional level, and as such, many tumor suppressor genes are found to be silenced in cancer. This Page 136 136 is NOT the case for oncogenes which are activated in cancer, not inactivated. Deacetylation of histone proteins is also implicated in gene silencing. Decitabine, an analog of 2’-deoxycytidine, is a drug that is both directly toxic to tumor cells and also appears able to affect cellular differentiation by re-activating silenced genes (including tumor suppressors) through demethylation. In Phase I/II clinical trials, it has shown some promise for the treatment of hematological malignancies. Rodríguez-Paredes M, Esteller M. Cancer epigenetics reaches mainstream oncology. Nat Med 17(3):330-9, 2011. PubMed Tsai HC, Baylin SB. Cancer epigenetics: linking basic biology to clinical medicine. Cell Res 21(3):502-17, 2011. PubMed Taby R, Issa JP. Cancer epigenetics. CA Cancer J Clin 60(6):376-92, 2010. PubMed Baylin SB, Jones PA. The next 10 years Timeline: A decade of exploring the cancer epigenome — biological and translational implications. Nat Rev Cancer 11(10):726-34, 2011. PubMed XV-4) E Active immunotherapy involves strategies designed to elicit the host’s immune system to attack tumor cells in vivo. The idea of manipulating a host’s immune system to attack a malignant tumor dates back more than 250 years. The success rate in the development of cancer vaccines in clinical trials had been low been low, until the development of Provenge. Patients undergo leukapheresis where peripheral blood mononuclear cells are cocultured with GM-CSF and the target antigen prostatic acid phosphatase (PAP). Cells are then transfused back into the patient intravenously. However, use of Provenge has been controversial, particularly in view of it’s high cost. One of the fundamental difficulties with cancer vaccines is that most tumor cells do NOT possess all the necessary characteristics to elicit a strong immune response and/or actively suppress their own immunogenicity. In model systems however, both T- and B-cells have been shown to be activated and attack tumor cells after priming with cancer vaccines. In addition, if tumor cells are first transfected with immunomodulatory cytokine genes, the immune response is strengthened. Moul JW. Putting Provenge in perspective. Oncology (Williston Park). 25(3):255, 258, 2011. PubMed Cheever MA, Higano CS. PROVENGE (Sipuleucel-T) in prostate cancer: the first FDA-approved therapeutic cancer vaccine. Clin Cancer Res 17(11):3520-6, 2011. PubMed Page 137 137 Emens LA. Cancer vaccines: on the threshold of success. Expert Opin Emerg Drugs 13(2):295-308, 2008. PubMed XV-5) B Nanotechnology involves the manipulation of atoms and molecules leading to the construction of nanometer scale objects or devices with unique properties suited for, among other things, the (earlier) detection of disease, monitoring of disease over time, and/or treatment of disease. Nanoparticles can be used for targeted drug delivery, biological testing for the presence or activity of specific molecular processes, medical imaging, and, in theory, repair of tissues damaged by disease or disease treatment. The National Cancer Institute is investing heavily in nanotechnology research as it relates to cancer. One advantage of using ultra-small scale nanodevices is that they can be specifically “engineered” to avoid biological or biophysical barriers. Grimm J, Scheinberg DA. Will nanotechnology influence targeted cancer therapy? Semin Radiat Oncol 21(2):80-7, 2011. PubMed Banerjee HN, Verma M. Application of nanotechnology in cancer. Technol Cancer Res Treat 7(2):149-54, 2008. PubMed Heath JR, Davis ME. Nanotechnology and cancer. Annu Rev Med 59:251-65, 2008. PubMed XV-6) C p16 INK4a normally inhibits the CDK4/6:cyclin D complex. Hence, its loss results in enhanced cellular proliferation. Li J, Poi MJ, Tsai MD. Regulatory mechanisms of tumor suppressor P16(INK4A) and their relevance to cancer. Biochemistry 50(25):5566-82, 2011. PubMed Musgrove EA, Caldon CE, Barraclough J, et al. Cyclin D as a therapeutic target in cancer. Nat Rev Cancer 11(8):558-72, 2011. PubMed Page 138 138 XV-7) D PTEN is a tumor suppressor gene whereas the others are all oncogenes. MYCN, HER2, MET and RET are amplified in neuroblastoma, breast and ovarian cancer, renal cell carcinoma and multiple endocrine neoplasia type II, respectively. Croce CM. Oncogenes and cancer. N Engl J Med 358(5):502-11, 2008. PubMed Cosgrove D, Park BH, and Vogelstein B. Tumor suppressor genes pp 86-101 In Cancer Medicine 8, Hong WK, Bast RC, Hait WN et al, Eds., B.C. Decker, Hamilton, 2010. Pierotti MA, Frattini M, Sozzi G and Croce C. Oncogenes pp 68-85 In Cancer Medicine 8, Hong WK, Bast RC, Hait WN, et al, Eds., B.C. Decker, Hamilton, 2010. Meyer N, Penn LZ, Reflecting on 25 years with MYC. Nat Rev Cancer 8(12):976-90, 2008. PubMed XV-8) B p53 (TP53) is defective in approximately half of human cancers. Phosphorylation of p53 by ATM is important in the G1/S cell cycle checkpoint. Phosphorylated p53 becomes a transcription factor, leading to expression of p21 (CDKN1A, WAF1/CIP1), which inhibits Cyclin E/CDK2. Giono LE, Manfredi JJ. The p53 tumor suppressor participates in multiple cell cycle checkpoints. J Cell Physiol 209(1):13-20, 2006. PubMed XV-9) A Although telomeres are maintained in germ line cells, most human somatic cells do not express sufficient telomerase to maintain telomeres on the ends of chromosomes, which shorten with each cell division. In contrast, cancer cells may have increased expression of telomerase which permits unlimited cell division to occur. Martínez P, Blasco MA.Telomeric and extra-telomeric roles for telomerase and the telomere-binding proteins. Nat Rev Cancer 11(3):161-76, 2011. PubMed O’Sullivan RJ, Karlseder J. Telomeres: protecting chromosomes against genome instability, Nature Reviews Molecular Cell Biology, 11:171-181, 2010. PubMed link Harley CB. Telomerase and cancer therapeutics. Nat Rev Cancer 8(3):167-79, 2008. PubMed Gilson E, Géli V. How telomeres are replicated. Nat Rev Mol Cell Biol 8(10):825-38, 2007. PubMed Page 139 139 XV-10) D RB (RB1) is frequently mutated in human cancer. RB and E2F are normally complexed together, which prevents entry of cells from G1/G0 into S phase. When conditions are right for movement from G1 into S, CDK's 4 and 2 are activated through binding with their cyclin partners, cyclins D and E, respectively. The CDK's then (hyper-) phosphorylate RB, causing a conformational change that make it detach from E2F. "Free" E2F then acts as a transcription factor for the actual genes required to move a cell from G1/G0 into S-phase. In tumor cells, either mutation of RB or elevation of cyclin D can lead to dysregulation of cell cycle progression. However, both conditions do not occur simultaneously. Udayakumar T, Shareef MM, Diaz DA, et al. The E2F1/Rb and p53/MDM2 pathways in DNA repair and apoptosis: understanding the crosstalk to develop novel strategies for prostate cancer radiotherapy. Semin Radiat Oncol 20(4):258-66, 2010. PubMed Burkhart DL, Sage J. Cellular mechanisms of tumour suppression by the retinoblastoma gene. Nat Rev Cancer 8(9):671-82, 2008. PubMed Knudsen ES, Knudsen KE. Tailoring to RB: tumour suppressor status and therapeutic response. Nat Rev Cancer 8(9):714-24, 2008. PubMed Macleod KF. The role of the RB tumour suppressor pathway in oxidative stress responses in the haematopoietic system. Nat Rev Cancer. 8(10):769-81, 2008. PubMed Page 140 140 XVI. Total Body Irradiation XVI-1) B Whole body irradiation is commonly used as part of the conditioning regimen for bone marrow transplantation. Such irradiation causes radiation damage to many tissues. Radiation-induced injury of the kidney demonstrates that lethal injury can result from doses used for transplant patients. The lung is also a concern particularly when the conditioning regimen includes cyclophosphamide and/or busulphan along with radiation. Faraci M, Barra S, Cohen A ,et al. Very late nonfatal consequences of fractionated TBI in children undergoing bone marrow transplant. Int J Radiat Oncol Biol Phys 63:1568-75, 2005. PubMed Sampath S, Schultheiss TE, Wong J. Dose Response and Factors Related to Interstitial Pneumonitis after Bone Marrow Transplant. Int J Radiat Oncol Biol Phys 63(3):876-84, 2005. PubMed Dawson LA, Kavanagh BD, Paulino AC, et al. Radiation-associated kidney injury. Int J Radiat Oncol Biol Phys 76(3 Suppl):S108-15, 2010. PubMed XVI-2) B Prolonged diarrhea, dehydration and loss of weight are seen after higher doses, approximately 8-10 Gy or more, that cause the gastrointestinal syndrome that is seen within days of exposure. A dose of 4 Gy to the whole body will result in the development of the hematopoietic syndrome, resulting from damage to the bone marrow. The clinical course following such a dose will be an initial prodromal syndrome, consisting of nausea, vomiting, abdominal cramps, fatigue etc. Since radiation will sterilize the mitotically- active precursor cells in the bone marrow, there will be a failure to replace mature blood cells lost from normal cell turnover. After irradiation, there will be a symptom free latent period of several weeks, after which the failure to replace the mature cells will result in a gradual depression of various blood cells, including granulocytes and platelets, resulting in the onset of chills, fatigue, and a decline in blood cell counts. Anemia is usually not observed. Williams JP, McBride WH. After the bomb drops: a new look at radiation-induced multiple organ dysfunction syndrome (MODS). Int J Radiat Biol 87(8):851-68. 2011. PubMed Mettler FA Jr, Gus’kova AK, Gusev I. Health effects in those with acute radiation sickness from the Chernobyl accident. Health Phys 2007 93(5):462-9. PubMed Page 141 141 XVI-3) E Individuals receiving an acute whole body dose of 10-20 Gy will die from the gastrointestinal syndrome. Currently, even aggressive medical care is of no value and only measures that provide comfort, such as pain management, administration of antiemetic and antidiarrheal agents, psychological support and pastoral counseling are advised. For people who receive less than 2 Gy, careful watching and response to specific symptoms would be most appropriate. It is unlikely transfusion will be necessary, and even if there is a drop in the blood constituents, this will not take place immediately. A person exposed to 5-10 Gy should be hospitalized under conditions that minimize infection, given antibiotics, receive cytokine therapy and be considered for a possible bone marrow transplant (at least in the higher end of this range). DiCarlo AL, Maher C, Hick JL, et al. Radiation injury after a nuclear detonation: medical consequences and the need for scarce resources allocation.Disaster Med Public Health Prep 5 Suppl 1:S32-44, 2011. PubMed Dainiak N, Gent RN, Carr Z, et al. First global consensus for evidence-based management of the hematopoietic syndrome resulting from exposure to ionizing radiation. Disaster Med Public Health Prep 5(3):202-12. PubMed Dainiak N, Gent RN, Carr Z, et al. Literature review and global consensus on management of acute radiation syndrome affecting nonhematopoietic organ systems. Disaster Med Public Health Prep 5(3):183-201, 2011. PubMed Wolbarst AB, Wiley AL, Nemhauser JB, et al.,Medical response to a major radiological emergency: A primer for medical and public health practioners. Radiology 254,660-677, 2010. Planning Guidance for Nuclear Detonation, first edition June 2010, Homeland Security Council Interagency Policy Coordination Subcommittee for Preparedness and Response to Radioological and Nuclear Threats. pdf link Medical Management of Radiological Casualties, Third Edition, 2009. pdf link Emergency Radiation Medicine Response—AFRRI Pocket Guide, 2008. pdf link ACR Disaster Preparedness for Radiology Professionals, A Primer for Radiologists, Radiation Oncologists and Medical Physicists, Government Version 3.0, 2006. pdf link Flynn DF, Goans RE. Nuclear terrorism: triage and medical management of radiation and combined-injury casualties. Surg Clin North Am 86(3):601-36, 2006. PubMed Page 142 142 Turai I, Veress K, Gunalp B, et al. Medical response to radiation incidents and radionuclear threats. BMJ 328(7439):568-72, 2004. PubMed Leikin JB, McFee RB, Walter FG, et al. A Primer for Nuclear Terrorism. Dis Mon 49(8):485-516, 2003. PubMed Moulder JE, Cohen EP. Future strategies for mitigation and treatment of chronic radiation-induced normal tissue injury. Semin Radiat Oncol 17(2):141-8, 2007. PubMed Waselenko JK, MacVittie TJ, Blakely WF, et al. Medical Management of the Acute Radiation Syndrome: Recommendations of the Strategic National Stockpile Radiation Working Group. Ann Intern Med 140(12):1037-51, 2004. PubMed XVI-4) C Prussian blue is ferric III hexacyanoferrate II which can be used to block the uptake of 137Cs in the intestines. Radioactive iodine uptake by the thyroid can be blocked by KI, but blocking of other agents is predominantly in experimental stages. XVI-5) E Potassium iodide decreases thyroid uptake of radioactive isotopes of iodine, which might be released in accidents or incidents involving a nuclear reactor. These isotopes would not be present after this incident, therefore administration of KI would provide no benefit in this case. Barnett DJ, Parker CL, Blodgett DW, et al. Understanding radiologic and nuclear terrorism as public health threats: preparedness and response perspectives.J Nucl Med 47(10):1653-61, 2006. PubMed Pellmar TC, Rockwell S. Priority List of Research Areas for Radiological Nuclear Threat Countermeasures. Radiat Res 163(1):115-23, 2005. PubMed Leikin JB, McFee RB, Walter FG, et al. A Primer for Nuclear Terrorism. Dis Mon 49(8):485-516, 2003. PubMed XVI-6) B Radiation-induced hypotension develops in people exposed to a whole body dose of 6-8 Gy. Following a whole body dose of 2 Gy there is relatively little medical treatment necessary even though nausea may be experienced. It is only in the dose range of 8-10 Gy in a single dose that a bone marrow transplant may improve survival. For doses less than approximately 8 Gy, a person is likely to survive with good nursing care and use of antibiotics and cytokines (e.g. G-CSF). Above 10 Gy, death is currently inevitable from GI syndrome. Page 143 143 XVI-7) D Although a whole body dose in the 8-10 Gy range may cause death due to a combination of gastrointestinal or hematopoietic syndrome, this would likely occur within approximately two months following the irradiation. Also, with sufficient stem cell support and antibiotic availability, many victims can survive both the hematopoietic and gastrointestinal syndromes produced by doses in this dose range. However, survival past two months is not a guarantee of long-term survival and the person may die from one of several possible late radiation effects, such as lung fibrosis. Cerebrovascular syndrome is not experienced at these relatively low doses and would have been seen within hours to days of exposure whatever the case. Although there will be localized skin problems, this dose is nevertheless low enough to allow for sufficient recovery of skin as a whole. Baranov AE, Selidovkin GD, Butturini A, et al. Hematopoietic Recovery after 10-Gy Acute Total Body Radiation. Blood 83(2):596-9, 1994. PubMed XVI-8) B Nausea and vomiting demonstrate a dose-related incidence and severity with doses >2 Gy inducing vomiting within 4 hours. Therefore, 4 hours is considered by many to be a triage point. Due to the relatively long lifetimes of red blood cells, anemia will not be seen until >1 month post-exposure, although the other conditions associated with hematologic deficits associated conditions (neutropenia, lymphocytopenia) will have a much earlier onset. Granulocyte numbers can show an increase, known as an abortive rise, possibly reflecting mobilization of reserves. Even though skin does show a dose response in the form of erythema, variability between individuals prevents its use as a dosimeter (although it could be used to assess localization of exposure). For whole body doses in the range of 2-5 Gy, the symptoms of the prodromal syndrome are followed by a symptom-free period known as the latent period. Even after exposure to very large doses sufficient to cause the cerebrovascular syndrome, victims can still experience a short latent time. XVI-9) C Therapeutic total body irradiation requires treatment of the entire body, and importantly, the bone marrow. Shielding of the entire gastrointestinal tract is not feasible and would likely compromise the therapeutic effect of radiation. In clinical settings, total body irradiation is delivered in a fractionated manner or at low dose rate, and this increases the tolerance for most normal tissues. Hematopoeitic stem cells are not known to repopulate the gastrointestinal tract. Page 144 144 Appelbaum FR, Badger CC, Bernstein ID, et al. Is there a better way to deliver total body irradiation? Bone Marrow Transplant 10 Suppl 1:77-81, 1992. PubMed XVI-10) C The LD50/60 (radiation dose that will result in lethality to 50% of a population at 60 days) for whole body irradiation is approximately 3-4 Gy. The LD50/60 may be slightly lower for the elderly and infirmed, and slightly higher for the young and healthy. Presumably, crew aboard this submarine are mostly young and healthy, and therefore the LD50/60 might be expected to be somewhat higher than 3-4 Gy. Moreover, in this scenario, only 10% of the crew survived. This suggests that a dose higher than the LD50/60 of 3-4 Gy was received by the crew. With antibiotics and nursing the LD50/60 could be expected to double, to approximately 6-8 Gy, but for the situation presented, these measures were not available. It is likely that no one would survive for two months following a whole body dose greater than 8-10 Gy since death from gastrointestinal syndrome within several weeks would be inevitable. XVI-11) C Following exposure to a whole body dose of <2 Gy, outpatient management may be appropriate with appropriate instructions about home care and observation. For patients receiving a dose of >2 Gy, hospital admission for observation, supportive care, transfusions, cytokine administration, and tissue typing for hematopoietic stem cell transplant may be appropriate. In this situation, either outpatient or inpatient management might be appropriate, but administering a graft with 100% likelihood of rejection would likely cause harm, and is therefore inadvisable. Dainiak N, Gent RN, Carr Z, et al. First global consensus for evidence-based management of the hematopoietic syndrome resulting from exposure to ionizing radiation. Disaster Med Public Health Prep 5(3):202-12, 2011. PubMed XVI-12) D Laboratory studies may be of value in an uncontrolled radiologic event. Serum TGF-beta and IL-1 may be cytokines that are released in response to radiation, but the association of dose and time of exposure have not been precisely characterized in humans. Serum creatnine could rise in response to kidney damage, although this might not happen for several weeks. Absolute neutrophil count would be expected to first rise and then fall after exposure, and therefore is not a reliable measure of exposure (especially if time of exposure is unknown). Absolute lymphocyte count is the most sensitive measure of radiation exposure, and is readily available in most laboratory services. Page 145 145 Mettler FA Jr, Voelz GL. Major radiation exposure--what to expect and how to respond. N Engl J Med 346(20):1554-61, 2002. PubMed XVI-13) B Hypotension can be caused by any number of pathophysiologic processes, and during a mass radiologic terrorism event following an explosion it is important not to forget that it can occur for reasons other than radiation exposure. Patients should be managed as if they have not been exposed to radiation, which includes assessment and management of traumatic injuries. While hypotension as a result of radiation exposure is a poor prognostic sign, its presence in the setting of a radiologic event should not distract the clinician from other more likely causes. Furthermore, for the scenario envisioned, radiation-induced hypotension would be very unlikely since exposure was to a low level of radioisotopes. Fry DE, Schecter WP, Hartshorne MF. The surgeon and acts of civilian terrorism: radiation exposure and injury. J Am Coll Surg 202(1):146-54, 2002. PubMed Page 146 146 XVII. Clinically Relevant Normal Tissue Responses to Radiation XVII-1) C There are no pathognomonic features observed in the irradiated kidney. Irradiation of the kidneys leads to radiation nephropathy, seen clinically some six months or more after irradiation as anemia, azotemia and an increased blood pressure. These functional changes are accompanied by changes in renal histology. Radiation nephropathy is associated with functional and morphologic changes. The latter consist of changes in all components of the nephron, including the glomerulus and the tubules. These morphologic lesions are progressive, leading to glomerulosclerosis, tubulointerstitial fibrosis, and ultimately renal failure. Cohen EP, Fish BL, Sharma M, et al. Role of the angiotensin II type-2 receptor in radiation nephropathy. Transl Res 150(2):106-15, 2007. PubMed Moulder JE, Fish BL, Cohen EP. Treatment of radiation nephropathy with ACE inhibitors and AII type-1 and type-2 receptor antagonists. Curr Pharm Des 13(13):1317-25, 2007. PubMed Cohen EP and Robbins ME. Radiation Nephropathy. Semin Nephrol 23(5):486-99, 2003. PubMed XVII-2) B Chronic lesions have classically been described in terms of demyelination, vasculopathies and necrosis, and are generally confined to the white matter. Radiation-induced brain injury can lead to both acute and chronic changes. The acute lesions include an increase in blood brain barrier permeability and edema, due in part to a rapid burst of apoptotic cell death, which is seen in some normal cells found in the brain, including endothelial cells. Interestingly, it has been noted that radiation-induced tissue changes are more frequently observed in AVM (arteriovenous malformation) patients than tumor patients, suggesting that dose-volume relationships should be studied separately in these two different patient groups. Kim JH, Brown SL, Jenrow KA, et al. Mechanisms of radiation-induced brain toxicity and implications for future clinical trials. J Neurooncol 87(3):279-86, 2008. PubMed Levegrün S, Ton L and Debus J. Partial Irradiation of the Brain. Semin Radiat Oncol 11(3):259-67, 2001. PubMed Tofilon PJ, Fike JR. The Radioresponse of the Central Nervous System: A Dynamic Process. Radiat Res 153(4):357-70, 2000. PubMed Page 147 147 XVII-3) A Based on the time of expression, radiation-induced CNS injury has been classified into three reactions: acute, early delayed, and late delayed. Acute injury is expressed in days to weeks after irradiation. Early delayed injury is seen 1-6 months after irradiation and includes transient demyelination with somnolence and Lhermitte’s syndrome after brain and spinal cord irradiation, respectively. White matter necrosis and cognitive impairment represent late delayed effects that are seen more than 6 months post-irradiation. There is a growing appreciation of the risk of cognitive impairment in long-term survivors. Although long recognized to be a major problem in children, where the severity of injury is inversely related with the age of the child at the time of irradiation, recent data show that cognitive impairment is a growing concern in adults as well. Johannesen TB, Lien HH, Hole KH, et al. Radiological and Clinical Assessment of Long-Term Brain Tumour Survivors after Radiotherapy. Radiother Oncol 69(2):169-76, 2003. PubMed Tofilon PJ, Fike JR. The Radioresponse of the Central Nervous System: A Dynamic Process. Radiat Res 153(4):357-70, 2000. PubMed Crossen JR, Garwood D, Glatstein E, et al. Neurobehavioral Sequelae of Cranial Irradiation in Adults: A Review of Radiation-Induced Encephalopathy. J Clin Oncol 12(3):627-42, 1994. PubMed XVII-4) D Skin irradiation leads to clonogenic cell loss in the stem cells present in the basal epidermis, with a resultant failure to replace the non-dividing, fully differentiated keratinocytes that continue to be sloughed off the skin surface at a normal rate. Hence, the latency period prior to the onset of moist desquamation is relatively independent of dose and is a reflection of the turnover time for the mature functional cells in the skin. At low doses, there is recovery from both surviving clonogens and adjacent areas, so little volume effect is seen. At higher doses, moist desquamation is normally seen at approximately 5 weeks after irradiation. Epilation is associated with doses greater than those required to initiate early transient erythema. The specific target cells involved in the acute and late effects observed after skin irradiation are distinct, and thus the severity of acute effects generally do not predict the severity of the late effects. Hopewell JW. The Skin: Its Structure and Response to Ionizing Radiation. Int J Radiat Biol 57(4):751-73, 1990. PubMed Page 148 148 XVII-5) D The radioprotector, amifostine, appears to be of benefit when present at the time of irradiation, serving as a classic radioprotector through its actions as a radical scavenger. It therefore does not provide any protective effect if used following irradiation. The use of IMRT to decrease the total volume of salivary gland tissue irradiated in head and neck cancer patients has been shown to markedly reduce the severity of radiation-induced xerostomia. Stringent oral hygiene is important due to (i) an increase in salivary viscosity leading to impaired lubrication and buffering capacity with increased risk for dental caries; oral flora become more pathogenic; (ii) failure to maintain oral hygiene will lead to plaque accumulation and risk of dental decay; (iii) the development of dental caries is accelerated in the presence of xerostomia due to a reduction in delivery of antimicrobial proteins normally present in saliva. A number of agents, including saliva substitutes and sialogogues, can provide palliative care. Redman RS. On approaches to the functional restoration of salivary glands damaged by radiation therapy for head and neck cancer, with a review of related aspects of salivary gland morphology and development. Biotech Histochem 83(3):103-30, 2008. PubMed Eisbruch A, Rhodus N, Rosenthal D, et al. The Prevention and Treatment of Radiotherapy - Induced Xerostomia. Semin Radiat Oncol 13(3):302-8, 2003. PubMed Chao KS, Majhail N, Huang CJ, et al. Intensity-Modulated Radiation Therapy Reduces Late Salivary Toxicity without Compromising Tumor Control in Patients with Oropharyngeal Carcinoma: A Comparison with Conventional Techniques. Radiother Oncol 61(3):275-80, 2001. PubMed XVII-6) C Many factors are accepted as affecting the development of osteoradionecrosis. These include the site of the primary tumor (mouth floor and oropharynx carrying the greatest risk), proximity of the tumor to the bone, and radiation dose. Tooth extraction and dental disease also have long been recognized as major risk factors. In addition, age, sex, nutritional status as well as alcohol and tobacco use may influence onset. Although the inclusion of the bone marrow sinusoids may affect the hematopoietic effects of irradiation, they appear to have no effect on the development of osteoradionecrosis. Wahl MJ. Osteoradionecrosis prevention myths. Int J Radiat Oncol Biol Phys 64(3):661-9, 2006. PubMed Teng MS, Futran ND. Osteoradionecrosis of the Mandible. Curr Opin Otolaryngol Head Neck Surg 13(4):217-21, 2005. PubMed Page 149 149 XVII-7) E Even in children less than 1 year of age, doses ≥5 Gy delivered as part of a standard fractionated protocol are generally required to induce cataracts. It also should be noted that recent studies suggest that the threshold dose for the development of the radiation-induced cataract has decreased to less than 1 Gy, possibly as low as 0.1 Gy, from the previously estimated threshold of 2 Gy. Neriishi K, Nakashima E, MinamotoA, et al. Postoperative cataract cases among atomic bomb survivors: radiation dose response and threshold. Radiat Res 168(4):404-8, 2007. PubMed Ainsbury A, Bouffler SD, Dörr W, et al. Radiation cataractogenesis: A review of recent studies. Radiat Res 172:1-9, 2009. PubMed XVII-8) A The α/β ratio for pericarditis has been estimated at roughly 2-3 Gy, indicating a large impact of fraction size. Classic studies performed in rabbits showed that the cells of greatest radiobiological importance in the heart are endothelial cells of the blood capillaries, whereas the cardiac myocytes are post-mitotic cells and are therefore much less radioresponsive. Large clinical studies on both Hodgkin’s and breast cancer patients have identified risk factors for the development of cardiac sequelae, and these include fraction size, dose and volume, with 2 separate trials indicating that age, particularly >60 years, has a strong influence. Adams MJ, Lipshultz SE, Schwartz C, et al. Radiation-Associated Cardiovascular Disease: Manifestations and Management. Semin Radiat Oncol 13(3):346- 56, 2003. PubMed Prosnitz RG, Chen YH, Marks LB. Cardiac toxicity following thoracic radiation.Semin Oncol 32(2 Suppl 3):S71-80, 2005. PubMed XVII-9) B In contrast to most radiation effects, the tolerance dose for sterility decreases with fractionation with the TD50/5 (the dose to produce the effect in 50% of people within a five-year period following irradiation of the particular organ/tissue) decreasing from approximately 6 Gy for a single dose to 2 Gy for a fractionated exposure. The reason for this unusual response is the long cell cycle time of Ap-spermatogonia and the presence of resistant populations as a result. Therefore, a single dose may not kill cells in the more resistant phases of the cell cycle. However, fractionation allows reassortment of these cells into radiosensitive portions of the cell cycle. In most other type- H tissues, the cell cycle time is much shorter and any enhanced effect due to Page 150 150 reassortment is counterbalanced by repopulation. Also, repair capacity, which normally counterbalances sensitization from reassortment appears to be low in spermatogonia. Testes are extremely sensitive to the effects of irradiation, with the radiobiological target identified as the type B spermatogonia. A dose of 1 Gy will result in a transient sterility due to azospermia; recovery is likely at about 1 year, although function is unlikely to return if sterility continues beyond 3 years. Although a permanent sterilizing dose occurs at approximately 5-6 Gy, total impairment of gonadal endocrine function does not occur until doses of 20-30 Gy are received. Hermann RM, Henkel K, Christiansen H, et al. Testicular Dose and Hormonal Changes after Radiotherapy of Rectal Cancer. Radiother Oncol 75(1):83-8, 2005. PubMed XVII-10) C The kidney does not appear capable of recovery from late functional damage. Preclinical data in rodents and pigs suggest that both radiation- induced acute and late skin injury can exhibit almost complete recovery. Experiments conducted in nonhuman primates (NHP) demonstrated a large capacity for recovery in the spinal cord; following an initial dose of 44 Gy in 2.2 Gy fractions only 4/45 NHP developed myeloparesis following retreatment with 57.2-66 Gy given 1-3 years after the initial radiation. Experimental studies in mice demonstrate recovery of re-irradiation tolerance in the lung for lethal pneumonitis. It appears likely that retreatment tolerance also occurs for late lung fibrosis, although it may be more limited compared with pneumonitis. Ang KK, Jian G-L, Feng Y, et al. Extent and Kinetics of recovery of occult spinal cord injury. Int J Radiat Oncol Biol Phys 50(4):1013-20, 2001. PubMed Terry NH, Tucker SL, Travis EL. Residual Radiation Damage in Murine Lung Assessed by Pneumonitis. Int J Radiat Oncol Biol Phys 14(5):929-38, 1988. PubMed Simmonds RH, Hopewell JW, Robbins ME. Residual Radiation-induced Injury in Dermal Tissue: Implications for Retreatment. Br J Radiol 62(742):915-20, 1989. PubMed Page 151 151 XVII-11) A The reduction in epithelial cell number takes place within one week following the start of radiotherapy in patients undergoing treatment for head and neck cancers whereas endothelial cell loss represents a late effect. While the early stage of oral mucositis is associated with mononuclear cell infiltration, there is generally no significant infiltration of neutrophils. The shorter the overall treatment time, the higher the incidence of (consequential) late effects after treatment of head and neck cancer. Kelland L. The resurgence of platinum-based cancer chemotherapy. Nat Rev Cancer 7(8):573-84, 2007. PubMed Rosenthal DI, Trotti A. Strategies for managing radiation-induced mucositis in head and neck cancer. Semin Radiat Oncol 19(1):29-34, 2009. PubMed Sonis ST. The Pathobiology of Mucositis. Nat Rev Cancer 4(4):277-84, 2004. PubMed Sonis ST, Elting LS, Keefe D, et al. Perspectives on Cancer Therapy-Induced Mucosal Injury: Pathogenesis, Measurement, Epidemiology, and Consequences for Patients. Cancer 100(9 Suppl):1995-2025, 2004. PubMed XVII-12) B Chronic radiation enteropathy is generally characterized by mucosal atrophy, rather than mucosal hypertrophy. The other features are common in chronic intestinal radiation injury. Wang J, Boerma M, Fu Q, et al. Significance of endothelial dysfunction in the pathogenesis of early and delayed radiation enteropathy. World J Gastroenterol 13(22):3047-55, 2007. PubMed XVII-13) C Pre-clinical data in monkeys demonstrate that the spinal cord has a large capacity to recover from prior radiation injury at a dose of 44 Gy in 2 Gy fractions. Only 4 of 45 monkeys developed myeloparesis after retreatment with 57.2 to 66 Gy given after 1-3 years after initial therapy. Although these data suggest that with sufficient time for repair, most of the spinal cord tolerance can be regained after radiation therapy with a standard dose and fraction schedule, in practice, many radiation oncologists discount only half of the prior radiation dose when considering the tolerance of the spinal cord to re-irradiation to maintain a margin for safety. Ang KK, Jiang GL, Feng Y. Extent and kinetics of recovery of occult spinal cord injury. Int J Radiat Oncol Biol Phys 50(4):1013-20, 2001. PubMed Page 152 152 XVIII. Mechanisms of Normal Tissue Radiation Responses XVIII-1) D Despite having cell turnover times that can be as long as months, changes in vascular endothelial cells, including alterations in the basement membrane and rough endoplasmic reticulum, have been seen within hours of irradiation. Fajardo LF, Berthrong M and Anderson RE, Eds., Radiation Pathology, Oxford University Press, New York, 2001 XVIII-2) E In attempts to individualize patient treatment, investigators have shown that high plasma levels of the profibrotic cytokine, TGFβ, both at the start of and, independently, continuing through the treatment period correlate positively with the later development of pneumonitis and fibrosis. The earliest responses seen in the lung following irradiation are apoptosis in the alveolar capillaries and release of surfactant, suggesting that the principal targets are the endothelial cells and the type II pneumocytes. However, the lung’s overall radiation response is unusual in some respects including the observation, seen both experimentally and clinically, that the response differs between inferior and superior irradiation fields. In addition, enormous individual variations have been seen in the response to radiation, particularly in the setting of bone marrow transplantation, making the identification of dose thresholds for different effects difficult to assess. A large number of drugs have appeared to increase the development of pulmonary late effects, although many have been through independent pathways. Brush J, Lipnick SL, Phillips T, et al. Molecular mechanisms of late normal tissue injury. Semin Radiat Oncol 17(2):121-30, 2007. PubMed Anscher MS, Vujaskovic Z. Mechanisms and Potential Targets for Prevention and Treatment of Normal Tissue Injury after Radiation Therapy. Semin Oncol 32(2 Suppl 3):S86-91, 2005. PubMed Rodemann HP, Blaese MA. Responses of Normal Cells to Ionizing Radiation. Semin Radiat Oncol 17(2):81-8, 2007. PubMed Page 153 153 XVIII-3) E Hepatic complications represent a major cause of morbidity and mortality in the bone marrow transplantation setting, particularly among pediatric patients, and symptoms typically occur within three months. Since the principal observed injury in the liver is vascular in nature (vascular occlusive disease), the radiobiological target has been hypothesized as the endothelial cell. The liver is deemed a flexible type of tissue and volume has been shown to have a significant effect on its response, with the tolerance (TD5/5) for liver complications estimated at 50 Gy, 35 Gy and 30 Gy following irradiation to 33%, 50% and 100% of the liver, respectively. Both total dose and fraction size have been shown to affect liver tolerance. Research to identify patients at risk of hepatic late effects following transplantation has identified high plasma levels of TGFβ as a positive indicator for increased probability of developing RILD. Dawson LA, Ten Haken RK. Partial volume tolerance of the liver to radiation. Semin Radiat Oncol 15(4):279-83, 2005. PubMed XVIII-4) B Functional sub-units (FSUs) are hypothesized to play a role in the volume effect seen in irradiated normal tissues. In clinical radiotherapy, the volume of tissue irradiated is an important factor determining the clinical tolerance of the organ. This can be very different from the radiosensitivity of the cells comprising the tissue per se. The kidney and lung are among the most radiosensitive organs when the total volume is irradiated. However, small volumes can be irradiated to much higher doses with little overall effect on organ function. The radiosensitivity of an organ depends on the arrangement of its FSUs and the ability of surviving, clonogenic cells to migrate from one FSU to another. In the kidney and lung, the FSUs are hypothesized to be arranged in a parallel manner. In the spinal cord, FSUs are hypothesized to be arranged serially. Loss of FSUs in a segment of the cord will lead to loss of function as the nerve impulse will no longer be able to be transmitted. The probability of this occurring will increase with the volume of cord irradiated. In addition, the clonogens within the FSUs in the spinal cord and skin are thought to be able to migrate over limited distances, accounting for the marked tolerance of these tissues to irradiation of small volumes. Since skin can recover from surviving clonogens and from adjacent areas, it does not exist in a serial arrangement and, at low doses, will demonstrate little volume effect. Page 154 154 XVIII-5) D Irradiation of normal tissues can result in chronic, progressive fibrosis. Radiation-induced fibrosis is now viewed as a dysregulated wound-healing response involving complex and dynamic interactions between several cell types rather than simply replacement of normal tissue by fixed fibrotic tissue. Experimental and clinical data indicate chronic production of a variety of inflammatory and profibrotic cytokines/growth factors produced by different cell types following irradiation of late responding tissues. In part, because of the multifactorial nature of the fibrotic response, studies during the last decade have shown that intervention in the steps comprising this process can partially reverse the development of fibrosis. Yarnold J, Brotons MC. Pathogenetic mechanisms in radiation fibrosis. Radiother Oncol 97(1):149-61, 2010. PubMed Kim K, McBride WH. Modifying radiation damage. Curr Drug 11(11):1352-65. PubMed Milano MT, Constine LS, Okunieff P. Normal tissue tolerance dose metrics for radiation therapy of major organs. Semin Radiat Oncol 17(2):131-40, 2007. PubMed Brush J, Lipnick SL, Phillips T, et al. Molecular mechanisms of late normal tissue injury. Semin Radiat Oncol 17(2):121-30, 2007. PubMed Fleckenstein K, Gauter-Fleckenstein B, Jackson IL, et al. Using biological markers to predict risk of radiation injury. Semin Radiat Oncol 17(2):89-98, 2007. PubMed Bentzen SM. Preventing or reducing late side effects of radiation therapy: radiobiology meets molecular pathology. Nat Rev Cancer 6(9):702-13, 2006. PubMed Anscher MS, Vujaskovic Z. Mechanisms and Potential Targets for Prevention and Treatment of Normal Tissue Injury after Radiation Therapy. Semin Oncol 32(2 Suppl 3):S86-91, 2005. PubMed Robbins ME, Zhao W. Chronic Oxidative Stress and Radiation-Induced Late Normal Tissue Injury: A Review. Int J Radiat Biol 80(4):251-59, 2004. PubMed Dent P, Yacoub A, Contessa J, et al. Stress and Radiation-Induced Activation of Multiple Intracellular Signaling Pathways. Radiat Res 159(3):283-300, 2003. PubMed Page 155 155 Williams J, Chen Y, Rubin P, et al. The Biological Basis of a Comprehensive Grading System for the Adverse Effects of Cancer Treatment. Semin Radiat Oncol 13(3):182-88, 2003. PubMed Denham JW, Hauer-Jensen M. The radiotherapeutic injury - a complex "wound". Radiother Oncol 63(2):129-45, 2002. PubMed XVIII-6) E It is now widely recognized that cytokines and growth factors play major roles in the development of radiation injuries to normal tissues. TNFα, IL-6, IL-1α and IL-1β are all important proinflammatory cytokines whose expressions are up-regulated within hours of irradiation. Their expression persists in a cyclical fashion until late effects are manifest. Interestingly, TNFα and IL-6 may play roles in tissue homeostasis due to their pro- and anti-apoptotic activities. TGFβ is upregulated both acutely and chronically following radiation injury, and its chronic expression is associated with progression to radiation- induced fibrosis. Although angiogenesis is an integral part of wound healing, VEGF does not appear to play a role in the induction of normal tissue late effects. Ikushima H, Miyazono K. TGFβ signalling: A complex web in cancer progression, Nat Rev Cancer 10(6):415-24, 2010. PubMed Schaue D, McBride WH. Links between innate immunity and normal tissue radiobiology. Radiat Res 173(4):406-17, 2010. PubMed . Deorukhkar A, Krishnan S. Targeting inflammatory pathways for tumor radiosensitizationBiochem Pharmacol 80(12):1904-14, 2010. PubMed Okunieff P, Chen Y, Maguire DJ, et al. Molecular markers of radiation-related normal tissue toxicity. Cancer Metastasis Rev 27(3):363-74, 2008. PubMed Fleckenstein K, Gauter Fleckenstein B, Jackson IL, et al. Using biological markers to predict risk of radiation injury. Semin Radiat Oncol 17(2):89-98, 2007. PubMed Bierie B, Moses HL. Tumour microenvironment: TGFbeta: the molecular Jekyll and Hyde of cancer. Nat Rev Cancer 6(7):506-20, 2006. PubMed McBride WH, Chiang CS, Olson JL, et al. A sense of danger from radiation. Radiat Res 162(1):1-19, 2004. PubMed Page 156 156 XVIII-7) A A major milestone achieved in recent years has been recognition by the scientific community of the role played by cytokines in radiation injury. Cytokines are proteins that regulate communications between cells. Many cytokines exhibit different activities on different cells, a characteristic known as pleiotropy. In addition, many functions are shared by different cytokines, an attribute known as redundancy. XVIII-8) A A stem cell is an undifferentiated cell that can produce progeny capable of either remaining stem cells, or committing to a pathway leading to differentiation. In keeping with their self-renewal ability for the entire lifetime of a tissue or organ, stem cells maintain telomerase activity. Stem cells are thought to reside in a special microenvironment, often referred to as a “niche”, with which they actively interact and can modify their reproductive behavior accordingly. The progeny of stem cells that are committed to differentiation are called progenitor cells or “transit amplifying cells”. These cells do not express telomerase, however they are usually rapidly- proliferating and expand the total population of cells while gradually differentiating and losing the ability to reproduce. Most, if not all, adult organs are now thought to contain stem cells, or at least to be able to produce stem cells in vitro; until recently, the prevailing view was that only select tissues (such as bone marrow, skin and several other epithelial tissues) contained stem cell compartments. For some tissues, the stem cells have yet to be identified unambiguously. It is likely that most human solid tumors likewise contain cancer stem cells that are also capable of self-renewal and differentiation, albeit poorly regulated. Some tumors also appear to recapitulate the [stem cell – committed progenitor cell – differentiated cell] sequence that occurs in their normal tissue of origin. Tumor stem cells are difficult to identify, and their response to traditional cancer therapies remains unclear. For example, it is possible that cell cycle-specific chemotherapeutics preferentially target the rapidly-proliferating progenitor cells, and not the stem cells themselves, that, relatively speaking, proliferate slowly. Soltanian S, Matin MM. Cancer stem cells and cancer therapy. Tumour Biol 32(3):425-40, 2011. PubMed Hittelman WN, Liao Y, Wang L, et al. Are cancer stem cells radioresistant? Future Oncol 6(10):1563-76, 2010. PubMed Pajonk F, Vlashi E, McBride WH. Radiation resistance of cancer stem cells: the 4 R's of radiobiology revisited. Stem Cells 28(4):639-48, 2010. PubMed Page 157 157 Visvader JE, Lindeman GJ. Cancer stem cells in solid tumours: accumulating evidence and unresolved questions. Nat Rev Cancer 8(10):755-68, 2008. PubMed Baumann M, Krause M, Hill R. Exploring the role of cancer stem cells in radioresistance. Nat Rev Cancer 8(7):545-54, 2008. PubMed Ishii H, Iwatsuki M, Ieta K, et al. Cancer stem cells and chemoradiation resistance. Cancer Sci 99(10):1871-77, 2008. PubMed Slack JMW. Origin of Stem Cells in Organogenesis. Science 322(5907):1498-501, 2008. PubMed Visvader JE, Lindeman GJ. Cancer Stem Cells in Solid Tumours: Accumulating Evidence and Unresolved Questions. Nat Rev Cancer 8(10):755-68, 2008. PubMed Cho RW, Clarke MF. Recent Advances in Cancer Stem Cells. Current Opin Gen Dev 18(1):48-53, 2008. PubMed XVIII-9) C Till and McCulloch were awarded the 2005 Lasker Prize for Basic Medical Research for their pioneering work in hematopoetic stem cell biology, effectively setting the stage for today’s research on adult and embryonic stem cells. Their work also led to the development of the “spleen colony assay”, a classic in vivo survival assay of radiosensitivity familiar to radiation oncology and biology. Both normal tissues and tumors are thought to contain stem cells capable of both self-renewal and differentiation, although for many tissues, these have yet to be identified. The progeny of stem cells that are committed to differentiation are called progenitor cells; they are usually rapidly-proliferating and expand the total population of cells while, in the case of normal tissues at least, gradually differentiating and losing the ability to reproduce. It has been suggested that the tumor equivalent of progenitor cells may be the ones most affected by traditional cancer therapies, whereas in fact, it is the slowly proliferating stem cells that should be considered the ultimate “tumor clonogen” requiring eradication. Many studies in recent years have demonstrated an unprecedented degree of “plasticity” of adult stem cells, including the ability to form cell types not native to their own tissues of origin. Although these findings remain controversial, clearly, adult stem cell therapy represents a tantalizing possibility. One possible application of adult (and/or embryonic) stem cells would be the prevention or amelioration of normal tissue damage caused by cancer therapy. Nuclear transfer is a controversial technology that allows the production of embryonic stem cell lines from adult somatic cells, including those from patients with genetically determined or influenced diseases. Once established in culture, such stem Page 158 158 cells could be manipulated using gene therapy to “correct” genetic abnormalities, and then returned to the patient (with which they would still be fully compatible) as a form of disease treatment. Weissman I. Stem Cell Research: Paths to Cancer Therapies and Regenerative Medicine. JAMA 294(11):1359-66, 2005. PubMed Mayhall EA, Paffett-Lugassy N, Zon LI. The Clinical Potential of Stem Cells. Curr Opin Cell Biol 16(6):713-20, 2004. PubMed Raff M. Adult Stem Cell Plasticity: Fact or Artifact? Annu Rev Cell Dev Biol 19:1-22, 2003. PubMed XVIII-10) D In hierarchically-organized tissues such as the intestine and oral mucosa, division and differentiation of stem and progenitor cells produce the mature cells that are responsible for maintaining tissue function. Loss of tissue function occurs as a result of insufficient replacement of functional cells. In such tissues, latency time (the time to expression of tissue injury) depends on cellular turnover rate (the faster the turnover rate, the shorter the latency time), but not on the radiation dose or on cellular radiation sensitivity. During fractionated radiation, there is usually significant repopulation and compensatory proliferation during treatment, and thus there is a strong dependence of tissue effect on overall treatment time. Page 159 159 XIX. Therapeutic Ratio XIX-1) D Any factor that could cause heterogeneity in the tumor would decrease the slope of the tumor control versus dose curve. Tumor heterogeneity can be caused by hypoxia, differences in the number of clonogens (which will affect tumor size) and tumor type and grade. Although the inherent radiosensitivity and volume of normal tissue in the field are also important to the extent that they could limit the total dose that can be given, they should not affect the slope of the tumor response curve. XIX-2) C In order for a treatment to result in a therapeutic gain, there must be an increase in the differential response between tumor and normal tissue, either making the tumor more sensitive without changing the normal tissue response, or protecting the normal tissue without protecting the tumor. Of the possibilities listed, only using a high LET ion beam with a tumor type having a large hypoxic fraction meets those criteria. Since the OER for high LET radiation is much lower than for X-rays, the hypoxic cells in the tumor will be more effectively killed than they would be the case for low LET radiation. XIX-3) B Generally, the radiation tolerance of normal tissues depends on the volume irradiated and the spatial organization of the functional subunits (FSUs). For the spinal cord, where the FSUs are arranged in series, loss of any one FSU has some probability of causing a complication. Hence, at small volumes (e.g., < 1 cm), the incidence of complications depends strongly on volume since undamaged cells from adjacent can migrate to the irradiated site and maintain function. However, if the radiation field is greater than approxiomately 1 cm, the distance is too great for viable cells from adjacent FSUs to repopulate the irradiated segment. XIX-4) E The critical factor is that the combination of the new drug with radiation must increase the therapeutic ratio. To benefit the patient, the new treatment must selectively increase the differential between tumor control and normal tissue toxicity resulting from treatment. If the response of the tumor and damage to the normal tissue both increase (or decrease) equally with the combination, the patients will not benefit. Page 160 160 XIX-5) D When preadolescents are irradiated, breast development is inhibited in 5% of the population at 10 Gy and in 50% of the population at 15 Gy. The risk of radiation-induced breast cancer is also increased in young girls and adolescents. XIX-6) B Even though the radiation may already be delivered at a low dose rate, fractionation produces further protection against radiation pneumonitis. Lung injury from chemotherapy, viral infection and smoking all increase the risk of pneumonitis. Because lung injury from graft-vs.-host disease also increases the risk of pneumonitis, allogeneic transplant recipients are at greater risk. XIX-7) B The tumor contains (10 7 ) x (0.01) = 10 5 clonogenic cells initially. However, during treatment delay the diameter of the tumor has doubled, meaning that the volume (and presumably, the number of clonogenic cells) has increased approximately 8-fold since volume is proportional to the cube of the diameter. Therefore, when treatment begins, the tumor contains roughly 10 6 clonogenic cells. For a 90% cure rate, there should only be a 10% probability that a single clonogen will survive and repopulate the tumor. Thus the radiation dose must produce a surviving fraction of 10 -7 . Since the effective D10 is 5 Gy, a dose of (7) x (5 Gy) = 35 Gy would be necessary to achieve a 90% cure rate. XIX-8) A Due to the exponential killing response produced by radiation, progressively fewer total cells are killed by each dose, although the fraction of cells killed is effectively the same. For example, if a tumor initially contained 1,000,000 cells and the dose per fraction resulted in a surviving fraction of 10%, then the first fraction would kill 900,000 cells, leaving 100,000 cells. The next dose would kill 90,000 cells leaving 10,000 cells, and so on. XIX-9) B Assuming that the tumor control probability (TCP) follows Poisson statistics, TCP is equal to e -(M)(SF) where M is the initial number of clonogenic cells and SF is the final surviving fraction for the total dose used in the treatment protocol. For a high SF (low levels of cell killing), the probability of tumor control would be extremely low and remain effectively zero over a large dose range. As the dose increases, eventually the TCP begins to increase and approach 100% as a maximum at relatively high doses (high cell killing and therefore low levels of SF), thus producing a sigmoidal response. In clinical practice, the increase in tumor control probability is not as steep as suggested by this equation due to tumor and treatment heterogeneity. Page 161 161 XIX-10) E Assuming that a solid tumor can be roughly modeled as a sphere, the volume of a sphere is proportional to the cube of its diameter. Assuming that the number of cells is proportional to the volume of a tumor, then the ratio of the number of cells in a 6 mm compared with a 2 mm diameter tumor is (6/2) 3 = 27. XIX-11) D To reduce 10 8 clonogenic cells to 10 2 , it is necessary to use a dose that would produce a surviving fraction of 10 -6 . Since 6 Gy results in a surviving fraction of 10 -1 , then (6) X (6 Gy) = 36 Gy would result in a surviving fraction of 10 -6 and reduce the number of clonogenic cells to 100. XIX-12) D Since 90% of the tumor cells were removed prior to irradiation, the dose to achieve the TCD50 can be reduced by one D10 value to a total of 60 Gy. Page 162 162 XX. Time, Dose, Fractionation XX-1) A None of the options is ideal, but delaying treatment for a short period is likely to be less detrimental than the other options for most solid tumors. Options B and D will increase the probability of normal tissue complications (NTCP) because fraction size is being increased for the normal tissue and NTCP is likely to be more affected by fraction size than tumor control probability (TCP). Options C and E will decrease TCP because the overall treatment time will be extended and this will increase the detrimental effect of accelerated repopulation occurring in the latter half of the treatment course. Huang J, Barbera L, Brouwers M, et al. Does Delay in Starting Treatment Affect the Outcomes of Radiotherapy? A Systematic Review. J Clin Oncol 21(3):555- 63, 2003. PubMed XX-2) B The rationale for switching to hyperfractionation is that radiation effects on late responding normal tissues will be spared preferentially relative to tissues or tumors that express radiation damage early after treatment. However, it is still critical to allow sufficient time between the fractions for full repair to occur. Half-times for repair in tissues are usually in the range of 1-1.5 hrs (spinal cord is an exception being 2 hours or more). Since roughly 4 half- times are required for what may be considered full repair, most authors recommend a minimum of about 6 hours between fractions of a hyperfractionated regimen. XX-3) D ARCON (accelerated radiotherapy, carbogen, nicotinamide) combines accelerated fractionated radiotherapy to counteract cellular repopulation, the hyperoxic gas carbogen (95-98% O2 + 2-5% CO2) to oxygenate hypoxic regions within the tumor and nicotinamide, a vasodilator to improve tumor blood flow and reduce perfusion-limited hypoxia. Hoskin P, Rojas A, Saunders M. Accelerated Radiotherapy, Carbogen, and Nicotinamide (ARCON) in the Treatment of Advanced Bladder Cancer: Mature Results of a Phase II Nonrandomized Study. Int J Radiat Oncol Biol Phys 73(5):1425-31, 2009. PubMed Jonathan RA, Wijffels KI, Peeters W, et al. The prognostic value of endogenous hypoxia-related markers for head and neck squamous cell carcinomas treated with ARCON. Radiother Oncol 79(3):288-97, 2006. PubMed Page 163 163 XX-4) B The α/β ratio for skin erythema is approximately 8-11 Gy. XX-5) E BED = nd x [1 + d/α/β] = 70 x [1 + (2/10)] =84 Gy10 XX-6) E The BED for the protocol described in option E is 117 Gy3, which is identical to the BED produced by the standard protocol.The BED values for options A through D, respectively, are 78, 100, 80 and 129 Gy3. But, without the need to perform calculations, A, B and D can be eliminated because they are not hyperfractionated patterns. Although option C describes a hyperfractionated protocol, since the total dose is lower than the standard protocol, it would not be possible for the BED values of the two protocols to be equal. XX-7) C Since carbon ions are high LET radiation, the survival curves for early and late responding tissues as well as for tumors would be expected to be nearly exponential. Accordingly, it would be anticipated that fractionation would have little impact. Thus, the isoeffect curves would be relatively flat as there would be little effect resulting from changing the dose per fraction. Late responding tissues treated with X-rays exhibit steeper isoeffect curves and greater dependence on dose per fraction than early responding tissues or tumors. The RBE value does not factor into determining the slopes of isoeffect curves. XX-8) E When patients are treated with more than one fraction per day in an effort to accelerate treatment, there is the risk of increased normal tissue toxicity since sufficient time may not be available between fractions to permit full repair of sublethal damage. This is a particular concern for the repair of damage associated with the development of late effects in the CNS. In fact, this problem has arisen in clinical trials, such as EORTC 22851, when only 4 hours were permitted between fractions for some patients. Both fraction size and overall time play an important role in determining acute effects. Hypofractionation involves the use of larger than traditional fraction sizes, but not necessarily a reduction in the total time to complete treatment. A split schedule permits recovery of early responding tissues, but may also lead to a loss of tumor control for more rapidly growing tumors. Fraction size and total dose are generally the most important factors affecting the probability of a late effect, whereas total treatment time plays a relatively minor role, unless the treatment is accelerated to such an extent that the acute responses are so severe that they develop into consequential late effects. Page 164 164 XX-9) A Irradiation with a series of small fractions, rather than a few large doses, generally reduces the risk of development of late effects caused by exposure to radiation. An accelerated protocol could reduce the amount of repair as there may not be sufficient time between fractions to allow full repair. To date, no agents have been identified that enhance repair capacity. Even if such an agent were available, it would not be certain whether this would reduce the risk of late effects for which the development of a perpetual cytokine cascade is probably of greatest importance. Irradiation under high oxygen tension will only enhance radiation damage in select hypoxic tissues. XX-10) D Although it is true that the use of small fraction sizes permits repair of sublethal damage in tumor cells, this enhances the survival of tumor cells and therefore does not represent a benefit. However, small fraction sizes of 1.8-2 Gy are typically used in radiotherapy since the sparing of normal tissues may be greater than the increased survival of cancer cells compared with the use of a large fraction size. In addition, reoxygenation of hypoxic tumor cells is thought to occur to a greater extent with the longer overall treatment times associated with the use of small dose fractions. XX-11) C Prolonging overall treatment time would likely have a larger sparing effect on early compared with late reactions since compensatory proliferation of early responding tissues may take place during treatment, whereas it is unlikely in late responding tissues. If treatment time is extended beyond the point when an early responding normal tissue begins to exhibit compensatory proliferation, the total dose may be increased without resulting in an increased severity of early reactions. However, this may be detrimental as it could result in an increased severity of late radiation reactions that do not benefit from compensatory proliferation. If the interval between fractions is decreased to less than 6 hours, the incidence of late normal tissue effects may increase due to incomplete repair. It is unlikely that patients with slowly growing tumors, i.e. those exhibiting Tpot values longer than 3 days, would benefit from accelerated treatment. In principle, it is patients with more rapidly proliferating cancers who would benefit from accelerated treatment. If the dose is titrated to maintain the same level of tumor control, then the use of a hyperfractionated schedule may result in a lower incidence of late effects. This is a principal reason for the use of hyperfractionation. XX-12) C All of the effects listed decrease following low dose-rate exposures, except possibly fetal effects induced during organogenesis. Even though low dose- rate irradiation may afford the opportunity for greater repair and recovery of fetal tissues, this is counterbalanced by the increased risk for congenital anomalies associated with the longer period of irradiation which may result in exposure of an organ that enters a critical period in its development during the course of irradiation. Page 165 165 XXI. Brachytherapy XXI-1) A Iridium-192 is currently the most widely used radioisotope for temporary implants. It has a number of advantages that include its small source size, lower photon energy that makes radiation protection less problematic, and usefulness with respect to computer-controlled remote afterloading. A disadvantage associated with the use of iridium-192, however, is that a range of dose rates will occur as the source decays (70 day half-life). Therefore, the dose administered must be adjusted for the dose rate. XXI-2) E As relatively short-lived isotopes used for brachytherapy, such as iridium-192 (70 day half-life) decay, both the time required to deliver a given total dose increases and the dose rate decreases over the course of treatment. The dose rate decrease is over a range where biological effectiveness changes rapidly due to the dose rate effect. Therefore, not only would an increase in treatment time be necessary to maintain the same total dose, but in fact the total dose should be increased to compensate for decreasing biological effectiveness as the dose rate drops. This concept was demonstrated clinically by Mazeron et al. in studies of local control and complication rates in head and neck cancers treated with interstitial iridium-192; for a given total dose, both local tumor control and probability of necrosis were less if the dose rate was less than 0.5 Gy/hour compared to 0.5 Gy/hour and above. Mazeron JJ, Simon JM, Le Pechoux C, et al. Effect of Dose Rate on Local Control and Complications in Definitive Irradiation of T1-2 Squamous Cell Carcinomas of Mobile Tongue and Floor of Mouth with Interstitial Iridium-192. Radiother Oncol 21(1):39-47, 1991. PubMed XXI-3) B Tumor control may increase if a total dose is delivered at a higher dose rate, although this could also increase damage to normal tissues that receive a significant radiation dose. A high dose rate would be beneficial if the tumor had a relatively low α/β ratio since the tumor cells would be more sensitive to changes in dose rate. Presumably, the accumulation of sublethal damage plays a large role in the killing of cells with a low α/β. Conversely, if the tumor is characterized by a relatively large α/β ratio, then treatment at a low dose rate would increase the therapeutic ratio since the late responding tissues, which likely possess a lower α/β, would be capable of repairing greater levels of sublethal damage resulting in diminished toxicity. Both I- 125 and Pd-103 emit relatively low energy photons, 28 keV and 21 keV respectively, which may be advantageous as their range is limited, thus resulting in lower doses to surrounding normal tissues. Page 166 166 XXI-4) A The β-particles produced by 90 Y are of a relatively high energy (900 keV). This provides a cross fire effect in which cells that did not take up label may still be irradiated. Pouget JP, Navarro-Teulon I, Bardiès M, et al. Clinical radioimmunotherapy-the role of radiobiology. Nat Rev Clin Oncol 8(12):720-34, 2011. PubMed Karagiannis TC. Comparison of different classes of radionuclides for potential use in radioimmunotherapy. Hell J Nucl Med 10(2):82-8, 2007. PubMed Macklis RM. Radioimmunotherapy as a therapeutic option for Non-Hodgkin's lymphoma. Semin Radiat Oncol 17(3):176-83, 2007. PubMed Hernandez MC, Knox SJ. Radiobiology of Radioimmunotherapy with 90 Y Ibritumomab Tiuxetan (Zevalin). Semin Oncol 30(6 Suppl 17):6-10, 2003. PubMed Page 167 167 XXII. Radiobiological Aspects of Alternative Dose Delivery Systems XXII-1) D Certainly the current cost for the production of carbon ions makes this form of treatment more expensive than X-rays. Densely ionizing radiations exhibit a reduction in the oxygen enhancement effect, making them valuable for the treatment of hypoxic tumors. Light ions, such as carbon, combine the high LET advantage of neutrons with the improved depth dose distribution of protons. Carbon ions deposit energy non-uniformly along the Bragg peak. The high LET portion of the Bragg peak can be placed in the tumor, sparing sensitive normal tissues around the tumor. Furthermore, the depth-dose profile of the carbon ion Bragg peak permits sparing at the entrance region of the beam. Okada T, Kamada T, Tsuji H, et al. Carbon ion radiotherapy: clinical experiences at National Institute of Radiological Science (NIRS). J Radiat Res (Tokyo) 51(4):355-64, 2010. PubMed Goitein M. Trials and tribulations in charged particle radiotherapy. Radiother Oncol 95(1):23-31, 2009. PubMed Jones B. The Case for Particle Therapy. Br J Radiol 79(937):24-31, 2006. PubMed Schulz-Ertner D, Jäkel O, Schlegel W. Radiation therapy with charged particles. Semin Radiat Oncol 16(4):249-59, 2006. PubMed XXII-2) C The main advantage to the use of protons is their advantageous dose distribution, enabling the delivery of a relatively large dose to a tumor when situated in the Bragg peak. In contrast, the biological properties of the high energy protons used for radiotherapy are similar to photons in that they are a low LET form of radiation, have a high OER and exhibit a similar variation in cell cycle radiosensitivity compared with photons. XXII-3) B Radiotherapy protons have an RBE of approximately 1.1 when compared with megavoltage photons. Page 168 168 XXII-4 D The late toxicities of neutrons were underestimated because of incorrect estimation of the RBE for critical tissues in patients. These RBEs were based on experiments that used large doses per fraction and measured early skin reactions in mice. In the clinical trials, the early skin reactions in patients receiving X-rays and neutrons were similar. However, the late reactions in patients treated with neutrons were much greater than anticipated because of the larger effect of fraction size for late compared with acute radiation responses (i.e. the α/β ratios for late effects are much lower than the α/β values for early skin effects). Since RBE is a ratio (X-ray dose to produce an effect divided by the test radiation dose to produce the same effect), the RBE used, which was based upon early radiation effects underestimated the RBE for late effects. Thus many of the patients treated with neutrons in the early trials developed intolerable late toxicities. Page 169 169 XXIII. Chemotherapeutic Agents and Radiation Therapy XXIII-1) C Human solid tumors are composed of two distinct but related compartments: malignant cells and supporting connective tissue (tumor stroma). Tumor stroma is composed of an extracellular matrix and a variety of cell types including immune/inflammatory cells, fibroblasts, vascular endothelial cells and perivascular cells. Growing evidence suggests that close interactions between the tumor and stromal compartments are critical in several steps of cancer development including tumorigenesis, tumor growth, and metastasis. Several lines of evidence suggest that agents targeting the tumor stroma may hold promise as cancer therapies. Bevacizumab is a monoclonal antibody directed against vascular endothelial growth factor and therefore is a stroma-targeted agent. Ibritumomab is a radiotherapeutic antibody directed against the B-cell antigen, CD20, and is used to target CD20+ malignant cells. Gemcitabine and cyclophosphamide are classic cytotoxic agents. Tirapazamine is a bioreductively activated DNA-damaging agent selective for hypoxic cells. The distinction between stroma-targeted and cancer cell-targeted agents is not entirely discrete as most cancer cell- targeted agents also have some effects on stromal cells and some stroma- targeted agents can directly affect cancer cells. Jain RK, Duda DG, Clark JW, et al., Lessons from Phase III Clinical Trials on Anti- VEGF Therapy for Cancer. Nat Clin Pract Oncol 3, 1:24-40, 2006. PubMed XXIII-2) C Both small molecule receptor tyrosine kinase inhibitors (TKIs) and monoclonal antibodies have been used in combination with chemotherapy and radiation therapy. TKIs are ATP-mimetics and include imatinib, gefitinib and erlotinib, among others. TKIs are generally administered orally. These agents show varied specificities against receptor tyrosine kinases and typically inhibit multiple targets at pharmacologically relevant concentrations. For example, imatinib (Gleevec) inhibits ABL, c-KIT and the PDGF receptor. In contrast, monoclonal antibodies display very high specificity and typically target a single receptor tyrosine kinase. Small molecule TKIs have been developed that inhibit receptor tyrosine kinases in both stromal and cancer cells. For example, sorafenib is a multi-targeted TKI that inhibits, among others, both tumor cell RAF kinase and stromal cell vascular endothelial growth factor receptor. Since the target of TKIs is the ATP binding domain of receptor tyrosine kinases as opposed to the ligand or the ligand binding domain typically targeted by monoclonal antibodies, these small molecule inhibitors are theoretically capable of inhibiting ligand-independent, constitutively active receptor tyrosine kinases. Page 170 170 Natoli C, Perrucci B, Perrotti F, et al. Tyrosine kinase inhibitors. Curr Cancer Drug Targets 10(5):462-83, 2010. PubMed Krause DS, Van Etten RA. Tyrosine Kinases as Targets for Cancer Therapy. N Engl J Med 353(2):172-87, 2005. PubMed XXIII-3) B Photodynamic therapy (PDT) uses a photosensitizing agent, oxygen and light of the appropriate wavelength to produce singlet oxygen (not hydroxyl radicals) that is toxic to cells. The efficacy of this approach depends on the ability of photosensitizers to localize preferentially in tumors. Because PDT requires oxygen, it is not effective against hypoxic cells. The range of activating light is usually 600-900 nm; at wavelengths less than 600 nm, tissue penetration is limited because of light absorption by endogenous molecules. In the clinic, the most commonly used photoactive agents are hematoporphyrin and its derivatives. Tirapazamine is a hypoxic cell cytotoxin and radiosensitizer, not a photosensitizing agent. Agostinis P, Berg K, Cengel KA, et al. Photodynamic therapy of cancer: an update Cancer J Clin. 61(4):250-81, 2011. PubMed XXIII-4) C The drug resistant phenotype, usually a consequence of gene amplification leading to changes in cellular DNA repair capacity and/or drug transport, has little, if any, bearing on sensitivity to ionizing radiation. Crespan E, Zucca E, Maga G. Overcoming the drug resistance problem with second-generation tyrosine kinase inhibitors: from enzymology to structural models. Curr Med Chem 18(19):2836-47, 2011. PubMed Cole S and Tannock I, Chapter 18. Drug Resistance. pp. 376-99, in The Basic Science of Oncology, 4th Ed. Tannock IF, Hill RP, Bristow RG, et al., Eds. McGraw-Hill, Medical Pub. Division, New York, 2005. XXIII-5) A On the assumption that drug resistant cells are already present in tumors prior to the initiation of chemotherapy, repeat exposures to the same drug(s) would tend to select for these resistant cells, ultimately leading to a loss of treatment response in the tumor as a whole. Therefore, the best way to avoid development of the drug-resistant phenotype would be to treat the tumor when it contains the fewest total cells, that is, when the presumed subset of resistant cells is likewise at its smallest. However, this is usually an impractical solution because even the smallest-sized tumor detectable with current imaging techniques probably already contains hundreds of millions of cells, including drug-resistant ones. Since the drug resistant phenotype is not Page 171 171 a cell cycle phenomenon per se, there would also be no benefit to using a non-cycle specific drug versus a cycle- or phase-specific one. And while multidrug resistance does tend to be greatest for chemotherapy agents derived from natural plant toxins, this is not exclusively the case, nor is multidrug resistance the only type of drug resistance overall. Finally, although calcium channel blockers such as verapamil or quinidine have been shown to reverse multidrug resistance in vitro, the doses required to accomplish this to a sufficient degree in vivo could result in intolerable toxicities. Therefore, of the answer options given, the best, if still not ideal, way to reduce the likelihood of a patient’s tumor becoming drug resistant would be to alternate courses of chemotherapy between two equally effective, but non-cross resistant, drugs. Theoretically, concomitant therapy may be even more effective, if the toxicity were tolerable. Boyer M and Tannock I. Chapter 17. Cellular and Molecular Basis of Drug Treatment for Cancer. pp. 349-75, in The Basic Science of Oncology, 4th Ed. Tannock IF, Hill RP, Bristow RG, et al., Eds. McGraw-Hill, Medical Pub. Division, New York, 2005. Cole S and Tannock I. Chapter 18. Drug Resistance. pp. 376-99, in The Basic Science of Oncology, 4th Ed. Tannock IF, Hill RP, Bristow RG, et al., Eds. McGraw-Hill, Medical Pub. Division, New York, 2005. Stewart F and Bartelink H. Chapter 20. The Combination of Radiotherapy and Chemotherapy. pp. 217-30, in Basic Clinical Radiobiology, 3rd Ed. Steel GG, Ed. Arnold, London, 2002. XXIII-6) D Drug-induced stimulation of repopulation would diminish the therapeutic index as this would increase the total number of tumor cells and therefore necessitate a larger radiation dose to achieve control. Begg AC, Stewart FA, Vens C. Strategies to improve radiotherapy with targeted drugs. Nat Rev Cancer 11(4);239-53, 2011. PubMed Strebhardt K, Ullrich A. Paul Ehrlich's magic bullet concept: 100 years of progress. Nat Rev Cancer 8:473-80, 2008. PubMed Helleday T, Petermann E, Lundin C. DNA repair pathways as targets for cancer therapy. Nat Rev Cancer 8(3):193-204, 2008. PubMed XXIII-7) C Redox reactions involving metals and O2 are directly involved in the chemical reactions that lead to DNA damage from bleomycin treatment. Page 172 172 XXIII-8) D In approximately 5% of patients with NSCLCs, interstitial deletion and inversion within chromosome 2p results in fusion of the N-terminal portion of the protein encoded by the echinoderm microtubule-associated protein-like 4 (EML4) gene with the intracellular signaling portion of the anaplastic lymphoma kinase (ALK) gene. EML4-ALK undergoes constitutive dimerization through interaction between the coiled-coil domain within the EML4 region of each monomer, thereby activating ALK and generating oncogenic activity. Tumor cells expressing EML4-ALK are “addicted” to its continued function. Crizotinib (Xalkori) is a small molecule inhibitor of the receptor tyrosine kinase activity of ALK. The EML4-ALK translocations are found predominantly in relatively young NSCLC patients who never smoked. It is estimated that roughly 10,000 new cases of NSCLC represent people with EML4-Alk translocations. Sunitinib (Sutent) is a small-molecule multiple receptor tyrosine kinase inhibitor of PDGFR-alpha, PDGFR-beta, VEGFR1, VEGFR2 and VEGFR3, KIT, FLT3, CSF-1R, and RET that is used to treat gastrointestinal stromal tumors (GIST) and renal cell carcinoma. Romidepsin (Istodax) is a histone deacetylase (HDAC) inhibitor approved to treat cutaneous T-cell lymphoma. Temsirolimus (Torisel) is an mTOR inhibitor used to treat renal cell carcinoma. Ipilimumab (Yervoy) is a recombinant, human monoclonal antibody approved to treat melanoma that binds to the cytotoxic T-lymphocyte-associated antigen 4 (CTLA-4), a molecule on T cells that suppresses the immune response. Blockade of CTLA-4 augments T-cell activation and proliferation and stimulates anti-tumor immune responses. Vemurafenib (Zelboraf) is used to treat melanomas that specifically possess the BRAFV600E mutation. Approximately 40 to 60% of melanomas carry mutations in the BRAF kinase that lead to constitutive activation of downstream signaling through the MAPK pathway. 90% of these mutations result in the substitution of glutamic acid for valine at codon 600(BRAF V600E). Mena AC, Pulido EG, Guillén-Ponce C. Understanding the molecular-based mechanism of action of the tyrosine kinase inhibitor: sunitinib. Anticancer Drugs 21 Suppl 1:S3-11,2010. PubMed Dancey JE, Curiel R, Purvis J. Evaluating temsirolimus activity in multiple tumors: a review of clinical trials. Semin Oncol 36 Suppl 3:S46-58, 2009. PubMed Culver ME, Gatesman ML, Mancl EE, et al. Ipilimumab: a novel treatment for metastatic melanoma. Ann Pharmacother 45(4):510-9, 2011. PubMed Minchom A, Young K, Larkin J. Ipilimumab: showing survival benefit in metastatic melanoma. Future Oncol 7(11):1255-64, 2011. PubMed Page 173 173 Shaw AT, Yasothan U, Kirkpatrick P. Crizotinib. Nat Rev Drug 10(12):897-8, 2011. PubMed Shaw AT, Yeap BY, Solomon BJ, et al. Effect of crizotinib on overall survival in patients with advanced non-small-cell lung cancer harbouring ALK gene rearrangement: a retrospective analysis. Lancet Oncol 12(11):1004-12. PubMed Chapman PB, Hauschild A, Robert C, et al. Improved survival with vemurafenib in melanoma with BRAF V600E mutation. N Engl J Med 364(26):2507-16. PubMed XXIII-9) A Doxorubicin is an antibiotic that directly binds to DNA and inhibits both DNA and RNA synthesis. XXIII-10) D The main dose limiting toxicity of doxorubicin is cardiac damage. XXIII-11) E The vinca alkaloids produce their main antitumor activity by binding to microtubule proteins and inhibiting microtubule polymerization. Komlodi-Pasztor E, Sackett D, Wilkerson J, et al. Mitosis is not a key target of microtubule agents in patient tumors. Nat Rev Clin Oncol 8(4):244-50, 2011. PubMed XXIII-12) B Methotrexate is a competitive inhibitor of dihydrofolate reductase. Page 174 174 XXIII-13) D Transfecting genes for patient-specific histocompatibility antigens into tumor cells would make them appear more “normal” to the host’s immune system. To enhance immunogenicity of tumors, it would be preferable to transfect genes that encode foreign antigens and/or genes encoding immune system stimulators. Addition of wild-type tumor suppressor genes, tumor-activated toxin genes and antisense oligonucleotides to oncogenes are other gene therapy strategies currently under investigation. Altaner C. Prodrug cancer gene therapy. Cancer Lett 270(2):191-201, 2008. PubMed Berinstein N. Chapter 21. Biological Therapy of Cancer. pp. 453-484, in The Basic Science of Oncology, 4th Ed. Tannock IF, Hill RP, Bristow RG, et al., Eds. McGraw-Hill, Medical Pub. Division, New York, 2005. XXIV-14) E Vorinostat (Zolinza) or suberoylanilide hydroxamic acid is a histone deacetylase inhibitor (HDACi) with activity against hemologic and solid cancers. Histones are part of the core proteins of nucleosomes. Acetylation and deacetylation of these proteins plays a role in the regulation of gene expression. HDACi can affect transcription by enhancing acetylation of histones, transcription factors, and other proteins regulating transcription. This is because acetylation relaxes the DNA packing around the histone core, making it accessible to transcription machinery. HDACi-related effects include growth arrest, cell death by apoptosis, autophagy and mitotic catastrophe, senescence, anti-angionesis, and ROS-facilitated cell death. Blumenschein GR Jr, Kies MS, Papadimitrakopoulou VA, et al. Phase II trial of the histone deacetylase inhibitor vorinostat (Zolinza, suberoylanilide hydroxamic acid, SAHA) in patients with recurrent and/or metastatic head and neck cancer. Invest New Drugs 26(1):81-7, 2008. PubMed Vorinostat (Zolinza) for cutaneous T-Cell lymphoma. Med Lett Drugs Ther 49(1256):23-4. PubMed Xu WS, Parmigiani RB, Marks PA. Histone deacetylase inhibitors: molecular mechanisms of action. Oncogene 26(37):5541-52, 2007. PubMed Page 175 175 XXIV-15) C Synthetic lethality is a promising approach to enhance cancer therapy option by targeting DNA repair proteins. A proof-of-principal for this approach has been realized with the discovery that inhibitors of the single-stranded break (SSB) DNA repair protein poly-(ADP)-ribose polymerase (PARP) are synthetic lethal in homologous repair (HR) deficient-cells with mutations in breast cancer susceptibility proteins, BRCA1 and BRCA2. In this case, when PARP1 is inhibited, SSBs persist, which leads to the collapse of replication forks in dividing cells and, ultimately, the formation of potentially lethal DNA double-strand breaks (DSB). In BRCA-deficient cells, the absence of HR which normally provides an error-free pathway to deal with replication fork associated lesions results in cell killing by concentrations of PARP inhibitor that are not toxic to HR-proficient cells. In addition to BRCA mutant-related cancers, PARP inhibitors such as ABT-888 (oliparib) and BSI 201 (iniparib) have activity against triple-negative breast cancers and several other solid tumors. Several ongoing clinical trials are investigating radiosensitizing effects of PARPi based on the radionale that some of radiation-induced SSB could convert to DSBs, if repair is inhibited by PARPi. Rouleau M, Patel A, Hendzel MJ, et al. PARP inhibition: PARP1 and beyond. Nat Rev Cancer 10(4);293-301, 2010. PubMed Rehman FL, Lord CJ, Ashworth A. Synthetic lethal approaches to breast cancer therapy. Nat Rev Clin Oncol 7(12):718-24, 2010. PubMed Chalmers AJ, Lakshman M, Chan N, et al. Poly(ADP-ribose) polymerase inhibition as a model for synthetic lethality in developing radiation oncology targets. Semin Radiat Oncol 20(4):274-81, 2010. PubMed Helleday T, Petermann E, Lundin C, et al. DNA repair pathways as targets for cancer therapy. Nat Rev Cancer 8(3):193-204, 2008. PubMed Kaelin WG Jr. The concept of synthetic lethality in the context of anticancer therapy. Nat Rev Cancer 5(9):689-98, 2005. PubMed Page 176 176 XXIV. Radiosensitizers, Radioprotectors and Bioreductive Drugs XXIV-1) A Studies have shown that inhibition of the large class of enzymes called histone deacetylases (HDACs) leads to an increase in radiosensitivity. This has been noted for numerous cell lines assayed both in vitro and in vivo. Some of the more common HDACi drugs are valproic acid, MS-275 and SAHA. A clinical biomarker for effective HDAC inhibition is to measure the extent of histone acetylation in circulating lymphocytes. This can likewise be measured in tumor biopsy specimens. Evidence for the effectiveness of HDACi drugs is the presence of hyper-acetylated histone proteins. Lane AA, Chabner BA. Histone deacetylase inhibitors in cancer therapy J Clin Oncol 27(32):5459-68, 2009. PubMed XXIV-2) D Activation by the liver is not required for cellular bioreduction of misonidazole to cause its conversion either to a hypoxic cell radiosensitizer or for any direct cytotoxicity this drug may have against hypoxic tumor cells. XXIV-3) B The hypoxic cell radiosensitizer nimorazole mimics the role of oxygen in radiation-induced free radical reactions, leading to radiosensitization; it does not increase oxygen concentration. Overgaard J. Hypoxic radiosensitization: adored and ignored. J Clin Oncol 25(26):4066-74, 2007. PubMed XIV-4) D Sulfhydryl compounds react with free radicals preventing these chemical species from reacting with and damaging critical cellular constituents. Kouvaris JR, Kouloulias VE, Vlahos LJ. Amifostine: the first selective-target and broad-spectrum radioprotector. Oncologist 12(6):738-47, 2007. PubMed Page 177 177 XXIV-5) D PARP inhibitors interfere with base excision repair (BER), leading to the formation of double strand breaks in S phase that require homologous recombination (HR) for their repair. In normal cells, PARP inhibitors are relatively non-toxic because functional HR is able to efficiently and accurately repair this type of damage. However, in patients with defective HR, such as those with mutations in BRCA1 and BRCA2 that occur in some breast, ovarian and pancreatic tumors, PARP inhibitors lead to double strand breaks that cannot be repaired and are thus lethal. Therefore, BER and HR are synthetically lethal in this context. Since BER is required for the repair of endogenously produced damage, the use of PARP inhibitors on their own in HR defective tumors is effective. No overlap in damage response between HR and NER, or MMR and BER is known. Amplification of receptor tyrosine kinases has been shown to stimulate repair and thus make cells more radiation resistant. Chan DA, Giaccia AJ. Harnessing synthetic lethal interactions in anticancer drug discovery Nat Rev Drug Discov 10(5):351-64, 2011. PubMed XXIV-6) D Hydroxyurea is an inhibitor of ribonucleotide reductase and kills S phase cells. Page 178 178 XXV. Hyperthermia XXV-1) C Treatment of cells at high temperatures leads to cell death primarily from protein denaturation. Griffin RJ, Dings RP, Jamshidi-Parsian A, et al. Mild temperature hyperthermia and radiation therapy: role of tumour vascular thermotolerance and relevant physiological factors.Int J Hyperthermia 26(3):256-63, 2010. PubMed Moyer HR, Delman KA. The role of hyperthermia in optimizing tumor response to regional therapy. Int J Hyperthermi 24(3):251-61, 2008. PubMed Dewhirst MW, Vujaskovic Z, Jones E, et al. Re-setting the biologic rationale for thermal therapy. Int J Hyperthermia 21(8):779-90, 2005. PubMed XXV-2) C Higher levels of cell killing yield higher levels of thermotolerance. Thermotolerance depends upon the level of cell killing, is not an inherited characteristic, takes longer than 10 minutes to decay and does not develop due to damage to mitochondria. XXV-3) A The increase in radiation sensitivity resulting from heat treatment is greatest when both are delivered simultaneously. Horsman MR, Overgaard J. Hyperthermia: a potent enhancer of radiotherapy. Clin Oncol (R Coll Radiol) 19(6):418-26, 2007. PubMed Kampinga HH, Dikomey E. Hyperthermic radiosensitization: mode of action and clinical relevance. Int J Radiat Biol 77(4):399-408, 2001. PubMed XXV-4) A Thermotolerance is associated with increased levels of molecular chaperones. Lepock JR. How do cells respond to their thermal environment? Int J Hyperthermia 21(8):681-7, 2005. PubMed Page 179 179 XXVI. Radiation Carcinogenesis XXVI-1) C The absolute risk model predicts the appearance of a distinct crop of radiation-induced cancers after a latent period following irradiation. This model did not predict a drop in the incidence of leukemia among Japanese A-bomb survivors. The relative risk model states that the risk of a radiation- induced cancer is a function of the natural incidence multiplied by a particular factor and it is this model that is favored by the BEIR VII committee to describe radiation-induced solid tumors. The overall radiation-induced cancer incidence among Japanese A-bomb survivors did NOT reach a plateau; excess cancers continue to rise even 60 years after the bombings. Health Risks from Exposure to Low Levels of Ionizing Radiation: BEIR VII Phase 2 (2006) National Research Council, National Academies Press, 2006. XXVI-2) A The younger the woman at the time of irradiation, the greater the probability of developing a radiation-induced breast cancer. The average latent period for radiation-induced leukemia is about 5-7 years. It remains unclear whether there is a threshold dose for cancer induction although the BEIR VII committee recommends use of the linear, no threshold (LNT) model. Radiation-induced solid tumors may appear decades after irradiation and generally arise at the same time as the natural incidence of that particular tumor type in the general population. Children are more susceptible to radiation-induced tumors compared with adults. Douple EB, Mabuchi K, Cullings HM, et al. Long-term radiation-related health effects in a unique human population: lessons learned from the atomic bomb survivors of Hiroshima and Nagasaki. Disaster Med Public Health 5 Suppl 1:S122-33. PubMed Review Article Newhauser WD, Durante M. Assessing the risk of second malignancies after modern radiotherapy. Nat Rev Cancer 11(6):438-48, 2011. PubMed Armstrong GT, Stovall M, Robison LL. Long-term effects of radiation exposure among adult survivors of childhood cancer: Results from the childhood cancer survivor study. Radiat Res 174(6):840-50, 2010. PubMed Shore RE. Low-dose radiation epidemiology studies: status and issues. Health Phys 97(5):481-6, 2009. PubMed Sadetzki S, Mandelzweig L. Childhood exposure to external ionising radiation and solid cancer risk. Br J Cancer 100(7):1021-5, 2009. PubMed Page 180 180 XXVI-3) E There is no evidence for an increase in prostate cancer incidence among the irradiated populations of Hiroshima and Nagasaki. Shore RE. Implications of radiation epidemiologic data for risk assessment and radiation protection. Health Phys 100(3):306-8. PubMed Preston DL, Cullings H, Suyama A, et al. Solid cancer incidence in atomic bomb survivors exposed in utero or as young children. J Natl Cancer Inst 100(6):428-36, 2008. PubMed Charles MW. LNT--An apparent rather than a real controversy? J Radiol Prot 26(3):325-9, 2006. PubMed Tubiana M, Aurengo A, Averbeck D, et al. The debate on the use of linear no threshold for assessing the effects of low doses, J Radiol Prot 26(3):317-24, 2006. PubMed Wakeford R, Little MP. Risk coefficients for childhood cancer after intrauterine irradiation: a review. Int J Radiat Biol 79(5):293-309, 2003. PubMed National Research Council (2006) Health Risks from Exposure to Low Levels of Ionizing Radiation: BEIR VII Phase 2. The National Academies Press, Washington, DC. Preston DL, Ron E, Tokuoka S, et al. Solid cancer incidence in atomic bomb survivors: 1958-1998. Radiat Res 168(1):1-64, 2007. PubMed XXVI-4) C A stochastic radiation effect is one in which the probability of the effect increases with dose and shows no apparent dose threshold. The severity of the effect is not related to dose. XXVI-5) E The incidence of breast cancer was elevated in patients who had received multiple fluoroscopies in connection with treatment for pulmonary tuberculosis. XXVI-6) E The number of people expected to develop a fatal cancer due to the radiation exposure is equal to the (number of people irradiated) X (average effective dose received) X (risk factor for the development of a fatal radiation-induced cancer) = (5 x10 6 people) X (2 x 10 -3 Sv) X (5 x 10 -2 fatal cancers/Sv) = 500 excess cancer cases. Page 181 181 XXVI-7) A The data obtained for the Japanese A-bomb survivors are consistent with a linear induction of solid tumors by radiation. Skin cancer appears to be a relatively uncommon form of radiation-induced cancer in the Japanese A- bomb survivors. Overall, the younger the person is at the time of irradiation, the greater is the risk for radiation-induced cancer. This is most apparent for radiation-induced breast and thyroid cancers. The lifetime risk of cancer fatality resulting from a whole body exposure among the general population for doses <0.2 Gy, or at a low dose rate, is approximately 5x10 -2 per Sievert. Leukemias represent about 20% of the fatal radiation-induced cancers among Japanese atomic-bomb survivors. Preston DL, Krestinina LY, Sokolnikov ME,et al. How much can we say about site- specific cancer radiation risks? Radiat Res 174(6):816-24, 2010. PubMed XXVI-8) D The severity of a deterministic effect increases with dose above a threshold, although not necessarily in a linear fashion. Normal tissue complications, which may have only a short latent period prior to their appearance, are examples of deterministic effects and are routinely produced in radiotherapy patients. Genetic effects are an example of a stochastic effect. XXVI-9) E Second cancers may be related to an underlying genetic predisposition or to a field effect from exposure to a carcinogen, such as cigarette smoking. Most people who develop Pancoast lung cancer have a smoking history, as is true for individuals who develop other forms of lung cancer. Therefore, in patients with a significant exposure to tobacco smoke, who develop head and neck cancer after being cured of lung cancer, the second malignancy is more likely to be the result of the tobacco smoke than prior radiotherapy. Radiation- associated second cancers can occur within or outside the prior radiation field. Radiation exposure outside of the radiation field may occur from internal scatter of photons and electrons within the patient or from radiation leakage emitted from the head of the gantry outside the radiation aperture. The latency of a radiation-related second cancer depends on the type of malignancy. Leukemias usually have a short latency, while the latency for solid tumors may be much longer. Page 182 182 XXVII. Heritable Effects of Radiation XXVII-1) C Based on animal studies indicating that the mutation incidence in offspring decreases if conception is delayed following irradiation, a period of 3-6 months before an attempted conception is recommended to patients undergoing a diagnostic procedure involving a significant dose to the gonads. Based primarily upon animal data, there is no evidence that a delay in conception of 3 years after a diagnostic procedure would decrease the chances of mutation compared to a 3-6 month delay. Laboratory studies indicate that even the very low doses used in diagnostic procedures are sufficient to induce heritable mutations in germ cells. XXVII-2) E Based on calculated risk estimates, approximately 1-6% of inherited mutations are due to exposure to background radiation. If humans were more sensitive than rodents to radiation-induced hereditary effects, then a statistically significant increase in mutations should have been detected among the the Japanese A-bomb survivors, however, this was not the case. It is assumed that mutagenesis is a stochastic effect of radiation, and therefore that a dose threshold for mutation induction does not exist. The dose to double the human spontaneous mutation rate has been estimated at 1 Sv. Radiation does not produce unique mutations that are not otherwise observed spontaneously. Fujiwara S, Suyama A, Cologne JB, et al. Prevalence of adult-onset multifactorial disease among offspring of atomic bomb survivors. Radiat Res 170(4):451-7, 2008. PubMed Boice JD Jr, Tawn EJ, Winther JF, et al. Genetic effects of radiotherapy for childhood cancer. Health Phys 85(1):65-80, 2003. PubMed Schull WJ. The children of atomic bomb survivors: a synopsis. J Radiol Prot 23(4): 369-84, 2003. PubMed Sankaranarayanan K, Wassom JS. Reflections on the impact of advances in the assessment of genetic risks of exposure to ionizing radiation on international radiation protection recommendations between the mid-1950s and the present. Mutat Res 658(1-2):1-27, 2007. PubMed Page 183 183 XXVIII. Radiation Effects in the Developing Embryo and Fetus XXVIII-1) E None of the procedures listed should result in a fetal dose greater than 0.1 Gy. This has been suggested as the threshold above which the physician should discuss the possibility of a therapeutic abortion with the patient due to the increased risk for congenital abnormalties. This would be especially true if the exposure took place prior to about 25 weeks of gestation. XXVIII-2) C Based upon rodent data, the greatest risk for congential abnormalties would be expected for irradiation during organogenesis, 1-6 weeks after conception, when most of the organ systems are developing. This is consistent with anecdotal human evidence for radiation-induced abnormalities. In addition, during the period of 8-15 weeks of gestation, there is a high risk (approximately 40%/Sv) for microcephaly and mental retardation. Irradiation during the first week post-conception during the preimplantation period carries little risk for the induction of congenital malformations as the irradiation would cause either a spontaneous miscarriage or else have no effect. The risk of congenital abnormality during either the second or third trimester is low, unless the fetal dose is very high. XXVIII-3) A During preimplantation (0-9 days) the embryo is sensitive to radiation- induced prenatal death. During organogenesis (1-6 weeks), there is an enhanced risk for neonatal death resulting from radiation-induced congenital malformations. For the early fetal period (6-15 weeks) there is an increased risk of mental retardation and microcephaly, for the middle fetal period (15-25 weeks) there would be a smaller risk of mental retardation, while in the late fetal period (20 – 40 weeks) there would be only an increased risk for cancer induction. Donnelly EH, Smith JM, Farfán EB, et al. Prenatal radiation exposure: background material for counseling pregnant patients following exposure to radiation. Disaster Med Public Health Prep 5(1):62-8, 2011. PubMed Preston DL, Cullings H, Suyama A, et al. Solid cancer incidence in atomic bomb survivors exposed in utero or as young children. J Natl Cancer Inst 100(6):428-36, 2008. PubMed Page 184 184 XXVIII-4) D Data obtained for Japanese A-bomb survivors who were irradiated in utero suggest that the main period of sensitivity for radiation-induced mental retardation is weeks 8-15 of gestation. A developing fetus appears to have approximately a 2-fold increased risk for cancer induction compared with the general population. The main abnormalities detected in this population were those of the CNS, presumably because of the relatively large period during gestation for CNS development. There is evidence consistent with the hypothesis that radiation exposure causes growth retardation. Although the threshold dose is not known precisely, radiation has been shown to produce measurable effects at doses less than 2Gy. Page 185 185 XXIX. Radiation Protection XXIX-1) C Approximately 3 millisieverts (mSv) per year is due to radiation exposure to the general public from natural sources such as the air, soil, building materials, etc. Greater than 50% of this dose is due to radon. XXIX-2) E Life-threatening injuries should always be treated before attempting decontamination of the patient. XXIX-3) Non-occupational radiation exposure: A. should not be permitted B. is allowed for education and training purposes C. should be limited to 10 mSv per year D. should be limited to 50 mSv per year E. should assume 1 mSv per year as the negligible individual dose XXIX-3) B Non-occupational exposure to radiation is permitted for educational and training purposes for people – even those younger than 18 years of age – but has to be limited to no more than 1 mSv per year. The negligible individual dose is 0.01 mSv per year. XXIX-4) D A radiation worker is permitted up to 50 mSv per year as long as the cumulative exposure limit of the worker’s age multiplied by 10 mSv is not exceeded. A 16 year old student performing research for a school project is permitted exposure up to 1 mSv per year. Members of the general public are allowed up to 1 mSv per year for continuous exposure and 5 mSv per year for infrequent exposure. A pregnant radiation worker is allowed 0.5 mSv per month of gestation once the pregnancy is declared. NCRP Report 116 Limitation of Exposure to Ionizing Radiation, 1993 Page 186 186 XXIX-5) D The collective dose is the product of the average dose and the number of individuals exposed. The committed dose is the dose resulting from incorporated radionuclides integrated over a 50 year period. The equivalent dose is the absorbed dose multiplied by a radiation weighting factor. The effective dose is the sum of equivalent doses to organs and tissue exposed, each multiplied by the appropriate tissue weighting factor. The genetically significant dose is the average dose equivalent to the gonads that is weighted for the age and sex distribution in those members of the irradiated population expected to have offspring. Thus, the GSD is not just the gonadal dose, but also reflects the average number of children it is anticipated that members of this population will have. XXIX-6) A If non-ionizing radiation modalities are available for diagnostic procedures and have equal or better sensitivity and specificity, then it may be beneficial to use such procedures to avoid the potential for radiation-induced mutations or cancers. However, for post-menopausal women who, by definition, are past their natural reproductive years, avoidance of ionizing radiation exposure will not affect the GSD, defined as the average effective dose to the gonads multiplied by the probability of conception. This assumes that menopause is accurately identified and is a definitive end to reproduction. “Surprise” babies in women who thought they were post-menopausal are not uncommon. XXIX-7) E Administration of potassium iodide soon after exposure (within 12-24 hours) to radioactive iodine would limit the uptake of this radioisotope by the thyroid. However, it must be kept in mind that an RDD may not disperse radioactive iodine, in which case use of potassium iodide will have no benefit. The presence of radioactive iodine can be determined rapidly using the equipment available to health physicists and first responders. It is unlikely that the detonation of a dirty bomb would result in many traumatic deaths. However, people who have sustained injuries should be treated in a hospital setting where they can obtain appropriate medical care. It is also unlikely that many people would receive sufficiently high doses to die from an acute radiation syndrome. In the event a terrorist does detonate a radiation dispersal device, it would be helpful for radiation experts to try to gain media access in order to help stem any panic and allay the fears of the population concerning radiation exposure. Accordingly, it would be wise for them to encourage the public within a 100 mile radius of the detonation who have not sustained physical injuries to avoid medical facilities and instead seek shelter in their offices and homes and to turn off any devices that may circulate contaminated air from the outside. Page 187 187 XXIX-8) B It has been estimated that as of 2007, medical procedures have surpassed radon as the largest single contributor to the average annual effective radiation dose received in the U.S. This is due primarily to the large increase in the use of diagnostic radiologic procedures, such as CT, which result in relatively large effective doses. In contrast, all of the other forms of radiation listed result in much smaller contributions to the average effective dose. NCRP Report No. 160, Ionizing Radiation Exposure of the Population of the United States Mettler FA Jr, Bhargavan M, Faulkner K, et al. Radiologic and nuclear medicine studies in the United States and worldwide: Frequency, radiation dose, and comparison with other radiation sources -- 1950-2007. Radiology 253(2):520- 31, 2009. PubMed Mettler FA Jr, Huda W, Yoshizumi TT, et al. Effective doses in radiology and diagnostic nuclear medicine: a catalog. Radiology 248(1):254-63, 2008. PubMed Amis ES Jr, Butler PF, Applegate KE, et al. American College of Radiology white paper on radiation dose in medicine. J Am Coll Radiol 4(5):272-84, 2007. PubMed Page 188 188 XXX. Molecular Techniques used in Radiation and Cancer Biology XXX-1) B Immunohistochemistry of phosphorylated H2AX phosphorylated (γ-H2AX) has become a powerful method for visualizing foci of a DNA repair-related protein in mammalian cells microscopically. Comet assay, filter elution assay and pulsed-field gel electrophoresis are all used to measure DNA break damage in whole cells or cell populations. The induction of p53 (TP53) is a response observed in cells that have been exposed to DNA damaging agents. Kuo LJ, Yang LX. Gamma-H2AX - a novel biomarker for DNA double-strand breaks. In Vivo 22(3):305-9, 2008. PubMed XXX-2) C The only assay of those listed that can be used to monitor gene expression is the cDNA microarray where cDNAs (or oligonucleotides matching cDNA fragments) are immobilized and then hybridized to RNA derived from various cell populations. The comet assay is used for monitoring DNA damage, the Southern blot is used for monitoring DNA-DNA hybridization, FISH records the chromosomal locations of various genes, and EMSA (electrophoretic mobility shift assay) monitors proteins that bind to DNA. XXX-3) C The Central Dogma is DNA>RNA>Protein. mRNA is first transcribed from nuclear DNA, processed, and then transported to the cytoplasm where ribosomes bind to it and translate it into a polypeptide chain. XXX-4) B Southern blotting involves transfer of denatured DNA from an agarose gel to a nitrocellulose filter where it can be hybridized with a complementary nucleic acid. It is therefore a DNA-based technique whereas gene expression is reflected in RNA levels, and all of the other techniques employ RNA to assess gene expression. XXX-5) D Electrophoretic mobility shift assay (EMSA) is used to identify the DNA sequence to which a transcription factor binds since DNA-protein complexes have a slower mobility than uncomplexed DNA in a gel. A far-western blot is a technique in which labeled proteins are hybridized to proteins that have been blotted to nitrocellulose. The two-hybrid screen is a genetic assay used to detect protein-protein interactions. Immunoprecipitation uses antibodies to bind to, precipitate, and thereby detect specific proteins. Page 189 189 XXX-6) C Restriction endonucleases are enzymes that cut DNA at specific palindromic sequences in DNA and they are widely used for cloning specific regions of DNA. XXX-7) D Northern blot analysis is a method to detect and determine the molecular weight of a messenger RNA (i.e., transcript) and to measure relative amounts of the mRNA present in either total RNA or an mRNA pooled sample. Western blotting (or protein immunoblotting) identifies with specific antibodies the molecular weight and relative amount of the protein present in different samples. Southern blotting, named after Edward M. Southern, allows an estimation of the the molecular weight of a restriction fragment to measure relative amounts in a DNA sample and to locate a particular sequence of DNA within a complex mixture. All DNA fingerprinting methods study patterns associated with genetic markers. Examples include a marker for a specific locus, the presence or absence of a restriction site in a DNA fragments. Southwestern blotting is used to study DNA-protein interactions. The membrane-bound proteins are incubated with a labeled double-stranded probe of specific DNA sequence. In combination with other techniques, this technique is useful to identification of potential protein factors. XXX-8) D The term “alkaline” provides an important clue to answer this question. Alkali treatment interferes with hydrogen bonding so the two DNA strands separate and can be analyzed independently for single-strand breaks. Page 190 190 XXX-9) E Choices A, B and E describe three different DNA arrays. The common feature of DNA array technology is that it permits simultaneous survey of a large number of DNA targets in a small sample and within a single hybridization experiment. A DNA array is a set of the ordered DNA sequences (targets or array elements) attached to a solid support. The 2-D Cartesian coordinates in the array define the identity of target DNAs. The common technical principle is hybridization probing, i.e., target sequences in the array identify complimentary sequences in the probe. In expression arrays, target DNAs are cDNAs, each representing a particular gene transcript. The hybridization probe is composite cDNA representing the total population of transcripts present in the cell. The main application of expression arrays is to compare the level of each mRNA in the array throughout the total mRNA population in the reference probe (generated from p53 (TP53)-wild type cells) versus that in the test probe (generated from p53- null cells). Comparative genomic hybridization (CGH) arrays are constructed from cloned DNA fragments from a specific candidate chromosomal region and are probed with total genomic DNA from the references and test samples. CGH arrays are used for genome-wide detection of chromosomal gains or losses. A polymorphism analysis array is used for genetic risk prediction. In this type of DNA array, DNA targets are mutational or polymorphic variants of a single gene and a probe is one DNA sample. Real- time reverse transcription polymerase chain reaction is useful for the analysis of a single gene transcript in multiple samples. Two-dimensional gel electrophoresis followed by sequencing is used to separate proteins by their molecular weight and charge, which may be followed by peptide sequencing or other protein-identification techniques. Page 191 191 XXXI. Molecular Imaging XXXI-1) C The magnet in an MRI system is described using a unit of magnetic flux density known as a tesla (T) which is equal to 10,000 gauss (G). The magnets in clinical MRI machines are in the 0.5-3.0 T or 5,000-30,000 G. Compared to the Earth’s 0.5 G magnetic field, a typical 1 T MRI magnetic field is approximately 20,000 times stronger than the earth’s magnetic field. XXXI-2) C A major difference between a computed tomography (CT) scan and positron emission tomography (PET) scan is that a CT scan gives anatomic information whereas a PET scan provides functional information. Due to the high contrast resolution of CT scanning, differences between the tissues are more apparent compared to other techniques. PET scans have the advantage over regular CT scans in that they provide information regarding metabolic activity and therefore can often differentiate between normal and abnormal tissues. The combination of CT and PET, or PET/CT scanners, provides coregistered images of anatomic and functional information in a single study (i.e., what the structure is, and what it is doing biochemically). Both procedures can increase the risk of cancer, because they use radiation. The resolution of these two techniques is similar. Bussink J, Kaanders JH, van der Graaf WT, et al. PET-CT for radiotherapy treatment planning and response monitoring in solid tumors. Nat Rev Clin Oncol 8(4):233-42, 2011. PubMed Brenner DJ, Hall EJ. Computed Tomography – An increasing source of radiation exposure. N Engl J Med 357(22):2277-84, 2007. PubMed Huang B, Law M, Khong PL. Whole-body PET/CT scanning: Estimation of radiation dose and cancer risk. Radiology 251(1):166-74, 2009. PubMed XXXI-3) B Hydrogen atoms have one proton and a large magnetic momentum and therefore can be used to create a “proton density map” of tissue properties (T1 and T2 weighting) and local magnetic field homogeneities (T1* weighting). Water is the greatest source of protons in the body, followed by fat. For this reason, MRI is best suited for imaging soft tissue. Bones are denser than the surrounding tissue but are virtually water-free. Thus, CT, but not MRI, is very good for imaging bony structures. Eye shadows often contain microscopic metal particles and can be a source of MRI image artifacts and/or increase in temperature and cause eye irritation. Page 192 192 Morgan B. Opportunities and pitfalls of cancer imaging in clinical trials. Nat Rev Clin Oncol 8(9):517-27, 2011. PubMed XXXI-4) A Positron emimtting radioisotopes are produced in the body along the proton path via nuclear reaction with carbon, oxygen, and nitrogen that can be detected on positron-emission tomography (PET) scan. The most abundant species of these positron emitters is carbon-11. In a matter of minutes, carbon-11 decays to boron-11, emitting a positron and a neutrino according to the equation: 11 6C → 11 5B + e + + νe + 0.96 MeV. In the proton therapy setting, PET can be used to verify proton range in the tumor, both during and after the proton irradiation. By acquiring a PET image during or right after treatment, the location of the positron emitters in the patient, and therefore the path of the proton beam, can be determined. By coupling the information from the PET image with the patient's anatomy, it is possible to monitor the location of the tumor and the location of the dose deposition. When positron- emitting isotope such as 11 C decays, the atomic number Z of the daughter isotope is one less that the parent, but the mass stays the same, because a proton becomes a neutron according to the equation: Energy + p → n + e + + νe. As it can be inferred from the above examples, proton-rich isotopes may decay via β-plus (e + ) positron emission (neutron-rich isotopes may decay via β-minus e - emission). Photoelectric absorption is negligible for 0.511 MeV photons resulting from the positron annihilation. The most important interaction for 0.511 MeV photons is Compton scattering. Parodi K, Enghardt W, Haberer T. In-beam PET measurements of beta+ radioactivity induced by proton beams. Phys Med Biol 47(1):21-36, 2002. PubMed Studenski MT, Xiao Y. Proton therapy dosimetry using positron emission tomography. World J Radiol 2(4):135-42, 2010. PubMed Ikotun OF, Lapi SE. The rise of metal radionucleotides in medical imaging: copper- 64, zirconium-89, and yttrium-86. Future Medicinal Chem 3(5):599-621, 2011. PubMed Zhou M, Zhang R, Huang M, et al. A chelator-free multifunctional [64Cu]CuS nanoparticle for simultaneous micro-PET/CT imaging and photothermal ablation therapy. J Am Chem Soc 132(43):15351-8, 2010. PubMed Page 193 193 XXXI-5) C Sir Godfrey Hounsfield is noted for developing CT and this scale was created to permit comparison of image opacity between images and scanners. XXXI-6) B The image intensities produced through CT can be used to predict dose deposition, which is useful for radiotherapy. MRI provides better tissue contrast than other techniques. Time of flight measurements are used in association with PET. CT is usually coupled with PET to permit improved image resolution. XXXI-7) B Agents such as nanoparticles containing gold that alter X-ray absorption can be used to improve on the minimal X-ray contrast between soft tissues. FDG and compounds containing positron emitters are used for PET. Iron- containing compunds are used for MRI and fluorescent molecules are used for detection using optical imaging approaches. XXXI-8) B FLT is a thymidine analogue that is phosphorylated by thymidine kinase, which is upregulated during DNA synthesis, which traps the radiotracer in proliferating cells. FDG is an analogue of glucose that accumulates in glycolytically active cells (tumor cells and inflammatory cells). FMISO, FAZA and EF5 are all radiotracers used to image hypoxic tissue. Bentzen SM, Gregoire V. Molecular imaging-based dose painting: a novel paradigm for radiation therapy prescription. Semin Radiat Oncol 21(2):101-10, 2011. PubMed Mankoff DA, Eary JF. Proliferation imaging to measure early cancer response to targeted therapy. Clin Cancer Res 14(22):7159-60, 2008. PubMed XXXI-9) C Dynamic imaging with CT after delivery of contrast agent can be used to image vasculature and quantify perfusion and vessel permeability. Hypoxia is not measured by CT. FDG-PET is utilized to quatify glycolysis.
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