LIFE09200604007 Tabish - Homi Bhabha National Institute

More documents

Recommendations

Info

FA along with ataxia telangiectasia, Bloom’s syndrome, and xeroderma pigmentosum are called cancer breakage syndromes because they are associated with chromosomal instability and defective repair, thereby predisposing to development of cancers. 15 Cells from FA show variable levels of cellular radiosensitivity, and results of an in vitro experimental study done for predicting the hypersensitive response may not coincide with in vivo radiosensitivity results. 21,22 Various experimental techniques including alkaline single-cell gel electrophoresis (Comet assay), colony-forming test, Western blot, and foci immunofluorescence analysis of the expression of DNA repair proteins, and also the cytochalasin-blocked micronuclei (MN) test using FA fibroblasts have been adopted to correlate the in vitro radiation sensitivity with the outcome of therapy. 23,24 However, the limitation of these techniques is the 4-week delay required for colony growth. 15,25–27 Comet assay is widely regarded as a robust and informative method for radiosensitivity measurements immediately after DNA damage, 28,29 but it is only a qualitative assessment of the response. Western blot and foci immunofluorescence analysis give qualitative results, but are labor intensive and also lack correlation between in vitro results and in vivo radiation response. 28 In view of the conflicting data on radiosensitivity of FA cells, and particularly FA fibroblasts, there is a pressing need for the development of new rapid and predictive assays of radiation responses. We have performed our study on PBMCs isolated from blood. Compared with most of the previous studies using fibroblasts, collecting blood has an advantage over invasive procedures of obtaining fibroblasts from patients’ skin biopsies, avoids time-consuming methods of generating monolayer cultures, and is also less labor intensive. Clinical radioresponsiveness is unpredictable in patients of FA with cancer, especially of the head and neck region, and outcome varies. 14 Of the 19 patients in the International FA Registry who had head and neck cancer, 14 died because of disease. The median time to recurrence in their patients was 16 months and the median follow-up of surviving patients was 19 months. 9 Kennedy and Hart 30 noted multiple primary malignancies either synchronous or metachronous in patients with FA to the extent of 36% (5 of 14 patients). Our patient developed a lymph node at level IV at 3 months follow-up and was alive with disease at 14 months. There are no standard guidelines for management of FA patients with malignancies. This is primarily a result of the scarcity of data and heterogeneous population. 13 It has therefore been recommended that more data in the form of case reports should be encouraged. 13 Until then, the treatment of patients with malignancies with FA should be individualized. Any decision about opting for different modalities should be based on a balanced approach with respect to locoregional control and toxicities of the treatment. REFERENCES 1. Alter BP, Young NS. The bone marrow failure syndromes. In: Nathan DG, Orkin SH, editors. Hematology of infancy and childhood. Philadelphia, PA: Saunders; 1998. pp 237–335. 2. Swift MR, Hirschhorn K. Fanconi’s anemia. Inherited susceptibility to chromosome breakage in various tissues. Ann Intern Med 1966;496–503. 3. Esparza A, Thompson WK. Familial hypoplastic anemia with multiple congenital anomalies (Fanconi syndrome): report of 3 cases. Cases presented are of two sisters and a female cousin with complete clinical and postmortem findings. R I Med J 1966;49:103–110. 4. Alter BP. Fanconi’s anemia and malignancies. Am J Hematol 1996;53:99–100. 5. Socie G, Socieux C, Gluckman E, et al. Squamous cell carcinomas after allogenic bone marrow transplantation for aplastic anemia: further evidence of multiple step process. Transplantation 1998;66:667–670. 6. Rosenberg PS, Greene MH, Alter BP. Cancer incidence in persons with Fanconi’s anemia. Blood 2003;101:822–826. 7. Auerbach AD, Allen RG. Leukemia and preleukemia in Fanconi anemia patients: a review of the literature and report of the International Fanconi Anemia Registry. Cancer Genet Cytogenet 1991;51:1–12. 8. Butturini A, Gale RP, Verlander PC, Adler-Brecher B, Gillio AP, Auerbach AD. Hematologic abnormalities in Fanconi anemia: an International Fanconi Anemia Registry Study. Blood 1994;84:1650–1655. 9. Kutler DI, Auerbach AD, Singh B, et al. High incidence of head and neck squamous cell carcinoma in patients with Fanconi anemia. Arch Otolaryngol Head Neck Surg 2003;129:106–112. 10. Oksuzoglu B, Yalcin S. Squamous cell carcinoma of the tongue in a patient with Fanconi’s anemia: a case report and review of the literature. Ann Hematol 2002;81:294–298. 11. Fukuoka K, Mishikawa K, Mizumoto Y, et al. Fanconi’s anemia with squamous cell carcinoma. A case report and a review of the literature. Jpn J Clin Hematol 1989;30: 1992–1996. 12. Lustig JP, Lugassy G, Sigler E. Head and neck carcinoma in Fanconi anemia: report of a case and review literature. Eur J Cancer B Oral Oncol 1995;31B:68–72. 13. Alter BP. Radiosensitivity in Fanconi’s anemia patients. Radiother Oncol 2002;62:345–347. 14. Bremer M, Schinder D, Gross M, et al. Fanconi’s anemia and clinical radiosensitivity report on two adult patients with locally advanced solid tumors treated by radiotherapy. Strahlenter Onkol 2003;179:748–753. 15. Marcou Y, D’Andrea A, Jeggo PA, Plowman PN. Normal cellular radiosensitivity in an adult Fanconi anemia patient with marked clinical radiosensitivity. Radiother Oncol 2001;60:75–79. 16. Bigelow SB, Rary JM, Bender MA. G2 chromosomal radio sensitivity in Fanconi’s anemia. Mutat Res 1979;63: 189–199. 1426 RT for Base of Tongue SCC in Fanconi’s Anemia HEAD & NECK—DOI 10.1002/hed October 2010
17. Taniguchi TD, D’Andrea A. The molecular pathogenesis of Fanconi anemia: recent progress. Blood 2006;107:4223–4233. 18. Kaplan MG, Sabio H, Wanebo HJ, Cantrell RW. Squamous cell carcinoma in the immunosuppressed patient: Fanconi’s anemia. Laryngoscope 1985;95:771–774. 19. Alter BP. Cancer in Fanconi anemia, 1927–2001. Cancer 2003;97:425–440. 20. Kutler DI, Singh B, Satagopan J, et al. A 20-year perspective on the International Fanconi Anemia Registry. Blood 2003;101:1249–1256. 21. Duckworth-Rysiecki G, Hulten M, Mann J, Taylor AMR. Clinical and cytogenetic diversity in Fanconi’s anemia. J Med Genet 1984;21:197–203. 22. Carreau M, Alon N, Bosnoyan-Collins L, Joenje H, Buchwald M. Drug sensitivity spectra in Fanconi’s anemia lymphoblastoid cell lines of defined complementation groups. Mutat Res 1999;435:103–109. 23. Arlett CF, Harcourt SA. Survey of radiosensitivity in a variety of human cell strains. Cancer Res 1980;40:926–932. 24. Djuzenova C, Flentje M, Plowman PN. Radiation response in vitro of fibroblasts from a Fanconi anemia patient with marked clinical radiosensitivity. Strahlenther Onkol 2004;180:789–797. 25. Finkelberg R, Thompson MW, Siminovitch L. Survival after treatment with EMS, X-rays, and mitomycin C of skin fibroblasts from patients with Fanconi’s anemia. Am J Hum Genet 1974;26:30A. 26. Weichselbaum RR, Nove J, Little JB. X-ray sensitivity of fifty-three human diploid fibroblast cell strains from patients with characterized genetic disorders. Cancer Res 1980;40:920–925. 27. Duckworth-Rysiecki G, Taylor AMR. Effects of ionizing radiation on cells from Fanconi’s anemia patients. Cancer Res 1985;45:416–420. 28. Djuzenova CS, Rothfuss A, Oppitz U, et al. Response to X-irradiation of Fanconi anemia homozygous and heterozygous cells assessed by the single-cell gel electrophoresis (Comet) assay. Lab Invest 2001;81:185– 192. 29. Mohseni-Meybodi A, Mozdarani H, Mozdarani S. DAN damage and repair of leukocytes from Fanconi anaemia patients, carriers and healthy individuals as measured by the alkaline comet assay. Mutagenesis 2009;24:67– 73. 30. Kennedy AW, Hart WR. Multiple squamous cell carcinomas in Fanconi’s anemia. Cancer 1982;50:811–814. RT for Base of Tongue SCC in Fanconi’s Anemia HEAD & NECK—DOI 10.1002/hed October 2010 1427
Page 1:
Genotype-Molecular Phenotype Correl
Page 7 and 8:
CONTENTS Page No Synopsis 1 List of
Page 9 and 10:
Synopsis
Page 11 and 12:
Synopsis Genotype-Molecular Phenoty
Page 13 and 14:
Synopsis Materials and Methods: 1.
Page 15 and 16:
Synopsis loading control. PCR produ
Page 17 and 18:
Synopsis 7.1 Percent cell death aft
Page 19 and 20:
Synopsis slides were then dried and
Page 21 and 22:
Synopsis range 41.77% - 90.95% at 5
Page 23 and 24:
Synopsis 5. Genotyping of genes: Ge
Page 25 and 26:
Synopsis G2/M arrest and delaying e
Page 27 and 28:
Synopsis may have an important bear
Page 29 and 30:
Synopsis References: 1. Bobba, R.;
Page 31 and 32:
Abbreviations MOPS : 3-(N-morpholin
Page 33 and 34:
List of Figures Fig. 28: Percent re
Page 35 and 36:
Chapter 1 Introduction
Page 37 and 38:
Introduction In previous studies fr
Page 39 and 40:
Introduction important carcinogenes
Page 41 and 42:
Review of Literature The yet undeci
Page 43 and 44:
Review of Literature Fig. 2: Incide
Page 45 and 46:
Review of Literature sustained angi
Page 47 and 48:
Review of Literature 2.6 Genotype p
Page 49 and 50:
Review of Literature 2.7.2 Biologic
Page 51 and 52:
Review of Literature Table 1: Steps
Page 53 and 54:
Review of Literature future analysi
Page 55 and 56:
Review of Literature Inefficient ce
Page 57 and 58:
Review of Literature XRCC1 (X-ray r
Page 59 and 60:
Review of Literature 2.9.2 Gene inv
Page 61 and 62:
Review of Literature MPO gene polym
Page 63 and 64:
Review of Literature important role
Page 65 and 66:
Materials and Methods Source of che
Page 67 and 68:
Materials and Methods Method: EBV-t
Page 69 and 70:
Materials and Methods Immunophenoty
Page 71 and 72:
Materials and Methods with 500 μl
Page 73 and 74:
Materials and Methods cDNA it can b
Page 75 and 76:
Materials and Methods Cobalt-60 iso
Page 77 and 78:
Materials and Methods specific bind
Page 79 and 80:
Materials and Methods h half of the
Page 81 and 82:
Materials and Methods phenol and th
Page 83 and 84:
Materials and Methods 7. Filter ste
Page 85 and 86:
Materials and Methods viz. EXO-SAP
Page 88 and 89:
Materials and Methods Table 2: List
Page 90 and 91:
Materials and Methods SMAD6 AREG HM
Page 92 and 93:
Materials and Methods 3.9 Effect of
Page 94 and 95:
Materials and Methods fresh eppendo
Page 96 and 97:
Materials and Methods Power SYBR Gr
Page 98 and 99:
Results Objective 1 To generate EBV
Page 100 and 101:
Results Fig. 8: Frequency of second
Page 102 and 103:
Results Table 6: Demographics of he
Page 104 and 105:
Results Fig. 10: Cell population do
Page 106 and 107:
Results Fig. 12: Cell surface marke
Page 108 and 109:
Results 4.1.5 Expression of ATM gen
Page 110 and 111:
Results Objective 2 To compare the
Page 112 and 113:
Results Fig. 17: Standardization of
Page 114 and 115:
Results Fig. 19: Standardisation of
Page 116 and 117:
Results The cells were incubated fu
Page 118 and 119:
Results 4.2.3.1 Percent G2 delay af
Page 120 and 121:
Results 4.2.3.2 Effect of BPDE expo
Page 122 and 123:
Results Fig. 29: Comparison of perc
Page 124 and 125:
Results Fig. 31: Percent cell death
Page 126 and 127:
Results normalization with baseline
Page 128 and 129:
Results 4.2.5.2 Real time PCR valid
Page 130 and 131:
Results homozygous wild-type, heter
Page 132 and 133:
Results Fig. 36b: Electrophoresis o
Page 134 and 135:
Results Fig. 37a: Representative el
Page 136 and 137:
Results Table 13: Consolidated geno
Page 138 and 139:
Results LCL Genes NAT2(1) NAT2(2) N
Page 140 and 141: Results Fig. 38: Distribution of Ge
Page 142 and 143: Results Fig. 39: Various combinatio
Page 144 and 145: Results Table 14: Genotype-phenotyp
Page 146 and 147: Discussion Introduction Squamous ce
Page 148 and 149: Discussion stimulated lymphocytes w
Page 150 and 151: Discussion H2AX is currently the mo
Page 152 and 153: Discussion or G2/M phase arrest. As
Page 154 and 155: Discussion differential expression
Page 156 and 157: Discussion DNA repair genes and MPN
Page 158 and 159: Chapter 6 Summary and Conclusions
Page 160 and 161: Summary and Conclusions Statistical
Page 162 and 163: Bibliography 1. Dikshit, R.; Gupta,
Page 164 and 165: Bibliography 43. Lei, D.; Sturgis,
Page 166 and 167: Bibliography 76. Fry, R. C.; Svenss
Page 168 and 169: Bibliography 115. Kote-Jarai, Z.; M
Page 170 and 171: Bibliography 149. Hashibe, M.; Bren
Page 172 and 173: Bibliography 183. Bergamaschi, D.;
Page 174 and 175: Bibliography 216. Bondy, M. L.; Wan
Page 176 and 177: Indian J Med Res 135, June 2012, pp
Page 178 and 179: 822 INDIAN J MED RES, JUNE 2012 mar
Page 180 and 181: 824 INDIAN J MED RES, JUNE 2012 (h)
Page 182 and 183: 826 INDIAN J MED RES, JUNE 2012 Tab
Page 184 and 185: 828 INDIAN J MED RES, JUNE 2012 lik
Page 186 and 187: CASE REPORT Eben L. Rosenthal, MD,
Page 188 and 189: FIGURE 2. Chromosomal breakage anal
show all

LIFE09200604007 Tabish - Homi Bhabha National Institute

Create successful ePaper yourself

Delete template?

Save as template?