Functional Evaluation and Interpretation of DNA Damage Repair Variants in Prostate Cancer

前列腺癌 DNA 损伤修复变异体的功能评估和解释

基本信息

批准号：
10708050
负责人：
CHARLES L. SAWYERS
金额：
$ 41.94万
依托单位：
SLOAN-KETTERING INST CAN RESEARCH
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-09-01 至 2025-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10708050
关键词：
Alleles Antineoplastic Agents BRCA1 gene BRCA2 gene Benign Biological Assay Biological Markers CHD1 gene CRISPR/Cas technology Cells Chromosomal Stability Clinical Clinical Research Clinical Trials Clustered Regularly Interspaced Short Palindromic Repeats Collaborations DNA DNA Repair Data Defect Disease Drug Exposure Engineering Evaluation Fanconi Anemia Complementation Group A Protein Fanconi Anemia-BRCA Pathway Gene Mutation Genes Genetic Genome Genomics Goals Grant Heterozygote Malignant neoplasm of prostate Mediating Missense Mutation Modeling Molecular Mutation Organoids Outcome PALB2 gene Pathogenesis Pathogenicity Pathway interactions Patients Pharmaceutical Preparations Phenotype Physiological Poly(ADP-ribose) Polymerase Inhibitor Prevalence Protein Truncation Radical Prostatectomy Recurrence Research Research Personnel Site Technology Testing Therapeutic Tumor-Derived Variant androgen deprivation therapy castration resistant prostate cancer design drug response prediction drug sensitivity experience gene repair genetic variant genome analysis genome editing genome sequencing genomic locus high risk insertion/deletion mutation loss of function mutation men mutant next generation sequencing participant enrollment patient population patient response prostate cancer cell prostate cancer cell line prostate cancer risk repaired response stability testing treatment response tumor whole genome

项目摘要

PROJECT SUMMARY/ABSTRACT Pathogenic variants in DNA damage repair (DDR) pathway genes are prevalent in a subset of men with metastatic castration-resistant prostate cancer (mCRPC) and occur in both the somatic and germline genomes. These abnormalities, primarily insertions and deletions resulting in protein truncations, occur in 10– 20% of patients with mCRPC. These variants also occur in lower-grade localized prostate cancers, although the prevalence of DDR mutations in these tumors and their impact on clinical outcome, survival, and drug sensitivity are largely unknown. In other projects of the proposed grant, specific mutations in DDR genes will be identified. The focus of Project 3 will be to prioritize and systematically evaluate these mutations, using a combination of functional assays, state-of-the-art organoid technology, CRISPR-mediated gene editing, and analysis of whole genome sequencing. In Aim 1 of Project 3, we will use multiple prostate cancer cell lines to determine whether specific DDR gene mutations are deleterious or benign variants. We will test the hypothesis that bona fide pathogenic DDR alleles are associated with a more aggressive tumor phenotype and will correlate with specific functional DNA repair defects and drug responses. In Aim 2, we will use gene editing to introduce DDR mutations into the endogenous genetic locus of multiple prostate cancer cell lines to provide a more physiologic assessment of the specific mutations. In Aim 3, we will analyze the whole-genome sequences from prostate cancers to identify precise mutational signatures which may result from loss-of- function mutations in specific DDR genes. Project 3 will have extensive collaborations with the investigators in Projects 1 and 2 and the Genomics Core as outlined in the attached research plan.

项目概要/摘要 DNA 损伤修复 (DDR) 通路基因的致病性变异在患有以下疾病的男性亚群中普遍存在转移性去势抵抗性前列腺癌 (mCRPC)，发生在体细胞和种系细胞中这些异常，主要是导致蛋白质截断的插入和缺失，发生在 10- 尽管 20% 的 mCRPC 患者也出现这些变异，但仍属于低级别局限性前列腺癌。这些肿瘤中 DDR 突变的患病率及其对临床结果、生存和药物的影响在拟议拨款的其他项目中，DDR 基因的特定突变的敏感性很大程度上是未知的。项目 3 的重点是使用以下方法对这些突变进行优先排序并系统地评估。功能测定、最先进的类器官技术、CRISPR 介导的基因编辑和在项目 3 的目标 1 中，我们将使用多种前列腺癌细胞系进行全基因组测序分析。确定特定 DDR 基因突变是有害还是良性变异，我们将检验该假设。真正的致病性 DDR 等位基因与更具侵袭性的肿瘤表型相关，并且将在目标 2 中，我们将使用基因编辑来关联特定的功能性 DNA 修复缺陷和药物反应。将 DDR 突变引入多个前列腺癌细胞系的内源遗传位点，以提供在目标 3 中，我们将分析全基因组。来自前列腺癌的序列，以识别可能因缺失而导致的精确突变特征项目 3 将与研究人员在特定 DDR 基因的功能突变方面进行广泛的合作。项目 1 和 2 以及基因组学核心如所附研究计划所述。