Xuechun Bai

Approximately 10% of bladder, upper urinary tract, and endometrial cancers harbor deleterious somatic alterations in nucleotide excision repair (NER) pathway genes. Although platinum-based chemotherapy is the standard treatment for NER-deficient tumors, its utility is limited by nephrotoxicity, motivating alternative strategies that avoid direct DNA targeting. Here, we employed a genome-wide CRISPR-Cas9 screen to identify synthetic lethal vulnerabilities in NER-deficient cancer cells. An ERCC4 -null RT112/84 bladder cancer cell line was generated and validated by Sanger sequencing and Western blotting, and functional NER deficiency was confirmed using a host cell reactivation assay. Parallel screens in ERCC4 -deficient and isogenic wild-type cells. nominated SLC25A28 as the top-ranked synthetic lethal candidate in ERCC4 -deficient cells, an interaction further supported by DepMap dependency. Clonogenic assays confirmed that loss of SLC25A28 selectively impaired cell viability in cells deficient for ERCC2 , ERCC3 , ERCC4 , or ERCC5 and inducible knockout selectively suppressed ERCC4 -deficient xenograft growth in-viv o. Mechanistically, RNA sequencing followed by gene set enrichment analysis revealed enrichment of oxidative stress-related pathways, including cellular responses to reactive oxygen species (ROS) and DNA damage signatures, upon SLC25A28 loss. These transcriptional changes are consistent with increased oxidative stress and DNA damage, placing heightened demand on NER-mediated repair. Direct measurement confirmed increased intracellular ROS. Collectively, our nominates SLC25A28 as a previously unrecognized synthetic lethal vulnerability in NER-deficient cancers and implicate mitochondrial iron homeostasis as a candidate metabolic dependency that may be targeted independently of platinum-based chemotherapy. Significance: This study identifies SLC25A28 as a synthetic lethal vulnerability in nucleotide excision repair–deficient cancers, linking mitochondrial iron homeostasis and oxidative stress to selective DNA damage intolerance and therapeutic opportunity.

Genetic similarity of populations (or genetic ancestry) is associated with differences in somatic alterations in cancers. We meta-analyze two targeted panel sequencing cohorts with 275,605 samples from 14 cancer types. Here we find a recurrent depletion of TERT promoter mutations in patients of African and East Asian ancestry across multiple cancers. Several clinically actionable alterations, such as ERBB2 mutations in lung adenocarcinoma and MET mutations in papillary renal cell carcinoma, occur at a higher frequency in patients of non-European ancestry. Furthermore, in both cohorts, we show depletions in total driver alterations in non-European ancestries in multiple cancer types, potentially reflecting biases in current panel-based testing that prioritize established targets derived from predominantly patients of European ancestry. Our study highlights a need to increase population diversity in genomic studies to find new drivers and enhance precision oncology interventions for all populations.

PURPOSE: In lung squamous cell carcinoma (LUSC), Black patients show significantly higher incidence and lower overall survival than White patients. Although socioeconomic factors likely contribute to this survival disparity, genomic factors have yet to be elucidated in LUSC. METHODS: Using 416 LUSC tumor samples in the Cancer Genome Atlas (TCGA), we assessed genomic and transcriptomic profiles by ancestry. We replicated our analyses in pan-cancer data from TCGA, the American Association of Cancer Research (AACR) Genomics Evidence Neoplasia Information Exchange (GENIE), and Columbia University Medical Center. RESULTS: We found increased MYC amplification, LUSC-specific MYC enhancer amplification, and chromosome arm 8q (chr8q) gain to be significantly associated with genetic AFR (African) ancestry in LUSC in TCGA. Furthermore, expression of MYC target genes was significantly enriched in AFR samples. Local ancestry analysis identified correlation of chr8q gain with AFR ancestry at the MYC locus in TCGA. We also found a significant correlation between chr8q and AFR ancestry in multiple cancer types and pan-cancer in TCGA. Similarly, in a pan-cancer subset of AACR GENIE data, we found a significant correlation between chr8q gain and race. CONCLUSION: Together, our data suggest that ancestry may influence amplification of not only MYC but also its enhancer in LUSC. They also suggest a role for genetic ancestry in chr8q aneuploidy in cancer. These studies further define and expand patients who may benefit from future anti- MYC therapeutic approaches.

Infectious diseases impact the swine industry through increased morbidity and mortality, leading to large economic losses and reduced animal welfare. Disease resilience is the ability of an animal to maintain production performance under pathogen exposure and has gained traction as an approach to help reduce the impact of infectious disease. For this purpose, a wean-to-finish polymicrobial natural disease challenge model was set up, in which, using a continuous flow system, batches of 60 or 75 young healthy Landrace × Yorkshire barrows from one of seven breeding companies of PigGen Canada were entered into a quarantine nursery before their entry in a nursery/finisher that was seeded with multiple pathogens. The objective of this study was to evaluate phagocytosis assays that can be conducted on blood from young healthy pigs as potential indicator traits to select for disease resilience in high-health nucleus breeding herds. Immune cell proportions and their phagocytic activity were evaluated in blood collected at ~26 days of age in the quarantine nursery on 985 pigs. Genetic parameters, including heritabilities and genetic correlations with disease resilience phenotypes collected on these pigs and an additional 2,220 pigs from the same companies in the polymicrobial challenge were estimated using GBLUP in ASReml 4.2, with the fixed effect of batch, the covariate of entry age, and random effects of pen by batch, litter, and additive genetics, using a genomic relationship matrix derived from genotypes of 650K single nucleotide polymorphisms. Results showed moderate heritability estimates for immune cell proportions (average = 0.36; range = 0.22-0.42) and the % of phagocytizing cells for each cell type (0.32; 0.19-0.48), but low estimates for mean fluorescence intensity (MFI) of the phagocytizing cells of each type (0.2; 0.05-0.3). Estimates of genetic correlations with disease resilience traits ranged from -0.93 to 0.72, but had substantial standard errors. Granulocyte % had a high positive genetic correlation with number of health treatments across the challenge nursery and finisher (0.72 ± 0.5), while a greater % phagocytizing eosinophils was genetically correlated with decreased growth rate in the challenge nursery (-0.26 ± 0.2) but greater growth rate (0.33 ± 0.2) and reduced mortality in the finisher (-0.78 ± 0.6) and across the challenge nursery and finisher (-0.50 ± 0.4). Greater MFI of phagocytizing eosinophils was also genetically correlated with reduced mortality across the challenge nursery and finisher (-0.93 ± 0.5). These results provide insights into potential indicator traits that can be used to select for disease resilience without the need to subject animals to disease, with the overall aim to improve animal welfare and increase production performance in herds that are subject to hard-to-manage infectious pathogens. This project was supported by Genome Canada, Genome Alberta, PigGen Canada, and USDA-NIFA grant #2017-67007-26144.