Chronic myeloid leukemia (CML), a clonal myeloproliferative disorder, is characterized primarily by the presence of chimeric BCR-ABL oncogene, and its progression from chronic to blast phase is associated with the accumulation of additional molecular and chromosomal abnormalities. The molecular mechanisms underlying this genetic instability are poorly understood. The activity of BCR-ABL is known to be associated with the increased production of intracellular reactive oxygen species and spontaneous DNA damage, which when effected by impaired/inaccurate DNA repair systems result in increased susceptibility to CML progression. Using case-control study design, we explored possible association of the repair gene, XRCC1, particularly the codons 399, 280, and 194 polymorphisms screened through PCR-RFLP, with the CML in the sample of 350 cases (206 male and 144 female) and 350 controls from Hyderabad, the capital city of state of the Andhra Pradesh, India. The patient group constituted 301 early chronic phase cases followed by 28 accelerated and 21 blast phase cases. The median age of the patients was 42 years (range, 9-70 years). The genotype distribution revealed significant association of codons 399 (χ(2) = 11.904, degree of freedom (d.f.) = 2; P = 0.002) and 194 (χ(2) = 8.091, d.f. = 2, P = 0.017) with CML, not 280 (P = 0.29). Although these polymorphisms are known to affect the function of XRCC1, the nature and extent of their genetic association with CML does not indicate their direct role in its development. The results seem to suggest that XRCC1 gene might have an important role in CML progression but not in its causation.

Abnormal apoptosis is one of the hallmarks of cancers including acute myeloid leukemia (AML), as it plays a pivotal role in precisely maintaining self-renewal, proliferation, and differentiation properties of hematopoietic stem cells (HSCs). Caspase9 (CASP9), an initiator caspase activated by mitochondrial-mediated apoptotic pathway (intrinsic pathway), triggers cascade of effector caspases and executes apoptosis. Functional SNPs in CASP9 might influence the gene expression leading to altered apoptosis which confer the risk to AML. To test this hypothesis, we have analyzed four CASP9 gene polymorphisms [CASP9 - 1263A > G (rs4645978), CASP9 - 712C > T (rs4645981), CASP9 - 293_275del CGTGAGGTC AGTGCGGGGA (-293del) (rs4645982), and CASP9 Ex5 + 32G > A (rs1052576)] in 180 AML cases and 304 age- and sex-matched healthy controls. We performed various statistical analyses to determine the potential interactions between these SNPs and AML. The study revealed that presence of G allele at CASP9 - 1263 position elevates the risk of AML 1.53-fold and CT/TT genotype at CASP9 - 712 position by 2.60-fold under dominant model of inheritance. Two CASP9 haplotypes, G-del(+)-C-A and G-del(+)-T-A, were found to be significantly associated with increased AML risk by 2.19- (95 % confidence interval (CI), 1.09-4.39; p = 0.028) and 11.75-fold (95 % CI, 1.01-136.57; p = 0.05), respectively. Further, multidimensionality reduction (MDR) analysis had revealed single locus CASP9 - 712C > T SNP and four loci CASP9 - 1263A > G, CASP9 - 293del, CASP9 - 712C > T, and CASP9 Ex5 + 32G > A SNPs as highest predicting models for AML development. Our results revealed a significant association of two SNPs in CASP9 (-1263A > G and -712C > T) and two haplotypes of the four SNP combinations with AML susceptibility.

Chronic myeloid leukemia (CML) is a monoclonal myeloproliferative disorder of hematopoietic stem cells (HSCs), characterized by reciprocal translocation, leading to the formation of BCR-ABL oncogene with constitutive tyrosine kinase (TK) activity. This oncogene is known to deregulate different downstream pathways which ultimately lead to cell proliferation, defective DNA repair, and inhibition of apoptosis. Fas (Fas cell surface death receptor) is a member of tumor necrosis factor (TNF) superfamily which interacts with its ligand, FasL, to initiate apoptosis. Promoter polymorphisms in Fas-FasL genes are known to influence the apoptotic signaling. Hence, the present study has been aimed to find out the association of the promoter polymorphisms in Fas and FasL genes with the development and progression of CML. Blood samples from 772 subjects (386 controls and 386 cases) were collected and genotyped for Fas-FasL gene polymorphisms through PCR-RFLP method. The association between SNPs and clinical outcome was analyzed using statistical softwares like SPSS version 20, SNPSTATs, and Haploview 2.1. The study revealed a significant association of Fas -670 G>A and FasL -844 T>C polymorphisms with the development of CML while Fas -670 AG was associated with accelerated phase. Combined risk analysis by taking the risk genotypes in cases and controls revealed a significant increase in CML risk with increase in number of risk genotypes (one risk genotype-OR 1.99 (1.44-2.76), p < 0.0001; two risk genotypes-OR 3.33 (1.91-5.81), p < 0.0001). Kaplan-Meier survival analysis of Fas -670 A>G and FasL -844 T>C showed reduced event-free survival in patients carrying the variant genotypes, Fas -670 GG, 32.363 ± 6.33, and FasL -844 CC, 33.489 ± 5.83, respectively. Our findings revealed a significant association of Fas -670 GG, FasL -844 TC, and CC genotypes with increased risk of CML.

B-cell lymphoma 2 (BCL2) and BCL2-associated X protein (BAX) proteins are anti-apoptotic and pro-apoptotic determinants of mitochondrial-mediated apoptosis, and their relative expression determines the cell fate. The promoter polymorphisms in these genes were shown to alter the protein function or expression and exert an impact on apoptosis regulation. Deregulation in the expression of any of these genes leads to disruption of cellular homeostasis and malignant transformation. The present study was aimed to determine the association of BCL2-938C>A and BAX-248G>A promoter polymorphisms with origin and progression of acute myeloid leukemia (AML). We also have performed combined genotype analysis to evaluate the cumulative effect of risk genotypes in the AML development. These polymorphisms were genotyped by polymerase chain reaction- restriction fragment length polymorphism (PCR-RFLP) in 221 AML patients and 305 age- and sex-matched healthy controls. Our study revealed that BCL2-938CA (p = 0.018) and BAX-248GG (0.043) genotypes were significantly associated with increased risk for AML occurrence. BAX-248A allele had shown decreased risk for AML. The combined analysis had shown that BCL2-938CA+AA-BAX-248GG group had a 1.63-fold (95 % CI: 1.08-2.45, p = 0.02) increased risk for AML. None of the clinical variables had shown any significant association with both polymorphisms. With respect to complete remission (CR) rate, BAX-248GG genotype (p = 0.002) and G allele (p = 0.009) had conferred significant risk for complete remission failure. Although the log rank test was not significant, survival analysis had shown a trend where BCL2-938CA genotype, and BAX-248GG had reduced median disease-free survival (DFS) of 9 and 10 months, respectively. In conclusion, BCL2-938C>A and BAX-248G>A gene polymorphisms might contribute to the origin of AML. Moreover, influence of BAX-248GG genotype on CR and DFS rate suggests that the BAX-248G>A polymorphism can serve as marker for poor prognosis in AML.

Background: The origin of Chronic Myeloid Leukaemia (CML) involves the formation of Double Strand Breaks (DSBs) which are initially sensed by the Ataxia Telangiectasia Mutated (ATM) signal kinase. Additionally, Bcr-Abl transformed CML cells produce high levels of Reactive Oxygen Species (ROS) that induce more DSBs. In such kind of cells, proper functioning of ATM and NHEJ repair mechanisms are required for ensuring the accurate repair of DSBs. Aim: The present study is planned to understand the role of DNA repair, specifically NHEJ repair in the occurrence of CML and its progression. Methodology: We have analysed seven genetic polymorphisms in ATM, XRCC5, XRCC6 and XRCC7 genes in 476 Ph+ve CML cases and 449 age and sex matched controls without family history of cancers. Direct Polymerase Chain Reaction (PCR) and Restriction Fragment Length Polymorphism (RFLP) analyses were carried out using the DNA isolated from blood samples through non-enzymatic/salting out method. Statistical analyses were performed through SPSS (IBM SPSS Statistics 20) software. Results: The T allele of ATM C4138T polymorphism and A allele of ATM -5144A&gt;T polymorphism were found to be significantly associated with enhanced risk of CML in our study. The CC genotype of XRCC6 -61C&gt;G polymorphism was found to have independent effect with CML risk as confirmed by the LD and MDR analyses. Lower repeat (0R) of XRCC5 VNTR and TT genotype of XRCC7 6721G&gt;T polymorphisms had shown borderline association with CML risk. Even though the XRCC5 2408G&gt;A and XRCC6 -1310C&gt;G polymorphisms did not show significant association with CML independently, the AA and GG genotypes of these polymorphisms have been found to confer enhanced risk for CML in combination with other SNPs of the respective genes in haplotype analysis. Interestingly, combined analysis demonstrated cumulative effect of risk genotypes in CML development as well as progression where the fold change increased with the number of adverse genotypes. Conclusion: The risk associated genotypes were known to influence the expression and functioning of genes in NHEJ pathway. Therefore, results of the present study suggested that defective DNA damage sensing and deregulated repair due to variations in ATM and NHEJ genes might influence CML development as well as progression. Key words: CML, ATM, NHEJ, DNA repair, polymorphisms Citation Format: Manjula Gorre, Prajitha Edathara Mohandas, Sailaja Kagita, Anuradha Cingeetham, Sugunakar Vuree, Sandhya Annamaneni, Raghunadharao Digumarti, Vishnupriya Satti. Genetic polymorphisms of DNA repair genes in the origin and progression of chronic myeloid leukemia. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 3858. doi:10.1158/1538-7445.AM2015-3858