RAD51 paralog function in cancer predisposition and genome integrity

RAD51 旁系同源物在癌症易感性和基因组完整性中的功能

基本信息

批准号：
10206963
负责人：
Kara A Bernstein
金额：
$ 44.68万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-03-15 至 2025-12-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10206963
关键词：
ATP phosphohydrolase Address Aging BRCA2 gene Biochemical Biological Biological Assay Biological Markers Bleomycin Cells Cellular Assay Chromosome abnormality Clinical Clustered Regularly Interspaced Short Palindromic Repeats Co-Immunoprecipitations Complex DNA DNA Damage DNA Double Strand Break DNA Repair DNA Repair Pathway DNA replication fork Development Double Strand Break Repair Embryo Environmental Carcinogens Environmental Exposure Exposure to Filament Functional disorder Genetic Genetic Polymorphism Genetic Screening Genomic Instability Germ-Line Mutation Goals Hereditary Breast and Ovarian Cancer Syndrome Human Human Cell Line Hybrids Individual Inflammatory Response Inherited Injury Ionizing radiation Knock-in Knock-out Knockout Mice Knowledge Link Malignant Neoplasms Malignant neoplasm of ovary Mammary Neoplasms Measures Mediating Metaphase Methods Microscopy Modeling Molecular Mus Mutate Mutation Ovarian Oxidative Stress PALB2 gene Pathogenicity Pathway interactions Proteins RAD51C gene Recombinant Proteins Reporter Role Screening for Ovarian Cancer Source System Technology Testing Therapeutic Time Toxic Environmental Substances Tracer Variant Visualization Walker-A Motif Walkers Woman Work XRCC2 gene XRCC3 gene Yeasts base brca gene cancer genome cancer predisposition cancer risk cost effective environmental agent environmental mutagens epidemiology study experimental study gene repair genome integrity high throughput screening homologous recombination in vivo individual patient malignant breast neoplasm member mutant next generation sequencing novel ovarian neoplasm oxidative damage paralogous gene precision medicine prevent protein protein interaction recruit repaired screening panel toxicant tumor variant of unknown significance

项目摘要

Project Summary/Abstract: Accurate repair of DNA damage is critical for genetic stability, and for preventing aging-related degeneration and cancer. We are working to identify key factors that regulate accurate repair of DNA double-strand breaks (DSBs) through the error-free homologous recombination (HR) pathway. DSBs can arise from many sources including endogenous replication fork damage, exogenous environmental toxicants, or oxidative stresses induced by endogenous sources and during pro-inflammatory responses to toxicant injury. We found that the RAD51 paralogs are critical for promoting HR and hence for suppressing error-prone repair mechanisms. Over 300 studies link mutations in human RAD51 paralogs with cancer, and women with breast or ovarian cancer are now screened for RAD51 paralog mutations. However, it remains largely unknown which RAD51 paralog mutations are pathogenic and how these mutations sensitize individuals to environmentally induced-DNA damage due to our lack of functional analysis of either the wild-type or mutated proteins. We do not know how these proteins are recruited, their functional components, or the disruptions caused by mutations or polymorphisms in the RAD51 paralogs. This knowledge gap results from low abundance of endogenous RAD51 paralog proteins, insolubility of the recombinant proteins, as well as embryonic lethality in knock-out mice. We are therefore using genetic, biochemical, and cell biological approaches to characterize RAD51 paralog function upon exposure to DSB inducing agents. We will use ionizing radiation (IR) and bleomycin as model agents for environmentally relevant DSB-inducing agents. Using complementary approaches in combination with high-throughput genetic screening, we are now uniquely poised to address how RAD51 paralog mutations predispose individuals to human cancer and thus, to identify opportunities for determining who is at risk for cancer development upon exposure to environmental carcinogens. Our ultimate goal is to enable development of precision medicine strategies for individual patients whose tumors harbor a RAD51 paralog mutation profile.

项目摘要/摘要： DNA损伤的准确修复对于遗传稳定性和预防与衰老相关的变性至关重要和癌症。我们正在努力确定调节DNA双链断裂准确修复的关键因素（DSB）通过无错误的同源重组（HR）途径。 DSB可以来自许多来源包括内源复制叉损伤，外源环境有毒物质或氧化应激由内源性来源以及对毒性损伤的促炎反应引起的诱导。我们发现 RAD51旁系同源物对于促进人力资源至关重要，因此对于抑制易于错误的修复机制而言。超过 300个研究将人Rad51旁系同源物与癌症或乳腺癌的女性联系起来现在筛选Rad51旁系同源物突变。但是，它在很大程度上仍然未知哪个Rad51旁系同源物突变是致病性的，这些突变如何使个体对环境诱导的DNA敏感由于我们缺乏对野生型或突变蛋白的功能分析而导致的损害。我们不知道如何这些蛋白质是募集的，其功能成分或突变或由突变或 RAD51旁系同源物中的多态性。这种知识差距是由于内源性低的丰度而引起的 RAD51旁系同源蛋白，重组蛋白的不溶性以及敲除的胚胎致死性老鼠。因此，我们使用遗传，生化和细胞生物学方法来表征RAD51 旁系同源物在暴露于DSB诱导剂后的功能。我们将使用电离辐射（IR）和博来霉素作为与环境相关的DSB诱导剂的模型代理。在结合高通量基因筛查，我们现在已经有了独特的准备来解决RAD51 旁系同源突变使人易于人类癌症，因此，确定了确定的机会在暴露于环境致癌物的情况下，他有癌症发展的风险。我们的最终目标是为肿瘤带有RAD51的个体患者制定精确医学策略旁同产突变曲线。