An analysis of likely germline events by tumor tissue testing on large somatic panels.
Authors: Bahadur Urvashi , Ravichandran Aarthi , Nayanala Swetha , Raju Sreelaksh , Muniyappa Nagesh , Krishnan Gayatri , G.R. Vijayashree , Chakraborty Vaishali , Pal Atanu , Noronha Vanita , Limaye Sewanti , Prabhash Kumar , ML Sheela , Ghosh Mithua , Pandey Rajeev Kumar , Phalke Sameer , Katragadda Shanmukh , Veeramachaneni Vamsi , Hariharan Ramesh 
Published: June 2024

e22524 Background: The increase in large panel testing has facilitated the detection of numerous events, including those in genes associated with inherited cancer. Further, large panel sequencing may uncover variants in these genes in non-inherited cancers. Hence the frequency of pathogenic/likely pathogenic (P/LP) events in some common germline genes was studied across cancers in the Indian population. Methods: A statistical model was used to derive a method to label variants as likely germline. Variants in hereditary breast and ovarian cancer (HBOC) and mismatch repair (MMR) genes from 1553 cases sequenced at our referral laboratory on the TruSight Oncology 500 panel (TSO500) were analyzed for their variant allele frequency (VAF) distribution. A 3 component Gaussian mixture model (intended to separate somatic, germline heterozygous, homozygous variants) was fit to this VAF distribution and a cutoff was determined such that the area under the curve of the somatic component above the cutoff was below 0.01. This gave a threshold value of 40% VAF for determining likely germline variants (LGVs). Data from 1713 TSO500 cases was then analyzed to identify LGVs. Results: The dataset of 1713 cases comprised 20.67% NSCLC, 12.61% breast, 12.49% ovarian and 10.5% colorectal cancer and < 10% of other cancer types including 2.39% pancreatic and 2.04% prostate cancer. 23.8% of ovarian, 17.14% of prostate, 12.5% of breast and 12.2% of pancreatic cancer cases had variants in BRCA1/BRCA2. Among these, 18.69% of ovarian, 17.14% of prostate, 8.33% of breast and 7.32% of pancreatic cancers had LGVs. LGVs in BRCA1/2 were also found in 3.33% of colorectal and 1.13% of NSCLC. The BRCA1/BRCA2 LGV ratio was found to be 3.0 in ovarian, 1.0 in breast and 2.0 in pancreatic but 0.2 in prostate and colorectal and 0.33 in NSCLC cancer. Adding other HBOC genes (ATM, BRCA1/2, BRIP1, CHEK2, PALB2, RAD51C, RAD51D) increased the LGV frequency to 22.43% of ovarian, 22.86% of prostate, 11.11% of breast, 9.76% of pancreatic, 4.44% of colorectal and 2.26% of NSCLC. MMR gene LGVs were seen in 1.23% of all cancers mainly in 10.71% of endometrial and 4.44% of colorectal cancers. Unlike the HBOC genes, MMR gene variants were not observed across multiple cancers and did not show a predominantly germline distribution (somatic/LGV 1:1). In contrast, the background P/LP frequency calculated from gnomAD v4 South Asian data (excluding those with conflicting labels) was 0.74% for BRCA1/2, 1.82% for the extended HBOC list and 0.5% for the MMR genes. Conclusions: Somatic testing was used to identify LGVs and determine the frequencies of HBOC and MMR genes in cancers in the Indian population. Somatic testing also identified LGVs in multiple tissues at frequencies above the background P/LP frequency for this population, suggesting a possible role for these genes in other cancers. This information underscores the need for further study and may be useful to define a clinically relevant subset of patients.