Abstract
Although genome-wide association studies (GWASs) have successfully revealed many common risk variants for bladder cancer, the heritability is still largely unexplained. We hypothesized that low-frequency variants involved in bladder cancer risk could reveal the unexplained heritability. Next-generation sequencing of 113 patients and 118 controls was conducted on 81 genes/regions of known bladder cancer GWAS loci. A two-stage validation comprising 3,350 cases and 4,005 controls was performed to evaluate the effects of low-frequency variants on bladder cancer risk. Biological experiments and techniques, including electrophoretic mobility shift assays, CRISPR/Cas9, RNA-Seq, and bioinformatics approaches, were performed to assess the potential functions of low-frequency variants. The low-frequency variant rs28898617 was located in the first exon of UGT1A3 and was significantly associated with increased bladder cancer risk (odds ratio = 1.50, P = 3.10 × 10−6). Intriguingly, rs28898617 was only observed in the Asian population, but monomorphism was observed in the European population. The risk-associated G allele of rs28898617 increased UGT1A3 expression, facilitated UGT1A3 transcriptional activity, and enhanced the binding activity. In addition, UGT1A3 deletion significantly inhibited the proliferation, invasion, and migration of bladder cancer cells and xenograft tumor growth. Mechanistically, UGT1A3 induced LAMC2 expression by binding CBP and promoting histone acetylation, which remarkably promoted the progression of bladder cancer. This is the first targeted sequencing study to reveal that the novel low-frequency variant rs28898617 and its associated gene UGT1A3 are involved in bladder cancer development, providing new insights into the genetic architecture of bladder cancer.
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Acknowledgements
This study was supported in part by the National Natural Science Foundation of China (81673264) and the Priority Academic Program Development of Jiangsu Higher Education Institutions (Public Health and Preventive Medicine). We are grateful to all the people who helped us accomplish this project. We thank Genesky Biotechnologies Inc. (Shanghai, China) for providing technical assistance.
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MW and ZZ designed the study and provided supervision. QL, CQ, YZ, CG, MW, and DY recruited study subjects. RZ, MD, YG, and JX performed statistical analyses and summarized results. FG, QZ, ZG, and SB performed functional experiments. RZ, MD, and HC prepared the manuscript.
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Zheng, R., Du, M., Ge, Y. et al. Identification of low-frequency variants of UGT1A3 associated with bladder cancer risk by next-generation sequencing. Oncogene 40, 2382–2394 (2021). https://doi.org/10.1038/s41388-021-01672-1
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DOI: https://doi.org/10.1038/s41388-021-01672-1
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