Investigating PolQ-mediated alternative NHEJ in breast cancer

研究乳腺癌中 PolQ 介导的替代 NHEJ

基本信息

批准号：
10090458
负责人：
Agnel Sfeir
金额：
$ 21万
依托单位：
NEW YORK UNIVERSITY SCHOOL OF MEDICINE
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：
2019-02-01 至 2021-03-03
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10090458
关键词：
Affect BRCA1 gene BRCA2 gene Biochemical Biotin Breast Cancer Model Cancer-Predisposing Gene Cells Characteristics Chemotherapy and/or radiation Chromosomal Rearrangement Chromosome abnormality Chromosomes Cicatrix Cisplatin Clinical DNA Damage DNA Double Strand Break DNA Repair DNA Repair Pathway Data Defect Development Disease Progression Double Strand Break Repair Enzymes Evolution Genes Genetic Genetic Recombination Genomic Instability Genomics Goals Human Ionizing radiation Knockout Mice Ligase Malignant Neoplasms Malignant neoplasm of ovary Mammalian Cell Mammary Neoplasms Mediating Molecular Mus Mutate Mutation Nonhomologous DNA End Joining Outcome Pathway interactions Patients Phenotype Polymerase Predisposition Proteome Proteomics Radiation therapy Relapse Resistance Role Shapes Signal Transduction Site Source System Testing Therapeutic Agents Treatment-Associated Neoplasms Work base brca gene breast cancer progression cancer cell cancer genome cancer therapy carcinogenesis chemotherapy cohort effective therapy established cell line experimental study genetic makeup genome integrity genome sequencing genomic signature homologous recombination improved in vivo inhibitor/antagonist insight malignant breast neoplasm mouse model neoplastic cell novel novel therapeutics ovarian neoplasm recruit repaired treatment strategy tumor tumor behavior tumor initiation tumor progression tumorigenesis whole genome

项目摘要

Chromosomal rearrangements are a hallmark of cancer cells and constitute a major pathway by which genes that affect tumor initiation and progression become mutated. Such aberrancies can result from defects in double-stranded break (DSB) repair. There are three major pathways of DNA repair in mammalian cells – the well-studied homology recombination (HR) and non-homologous end joining (NHEJ) pathways, and the poorly characterized, yet highly error-prone alt-NHEJ (alternative-NHEJ) pathway. Genetic rearrangements consistent with alt-NHEJ have been noted both in spontaneous and therapy-related tumors. In this proposal we will focus on polymerase theta (Polθ), a low-fidelity enzyme that we recently identified as a key factor that mediates DSB repair by alt-NHEJ. Given that HR-defective tumors are “addicted” to repairing DSBs via the alt-NHEJ pathway, we hypothesize that the mutagenic activity of Polθ help establish a genomic landscape that is conducive for aggressive tumor behavior. Additionally, we predict that deleting Polθ in tumors with mutations in the breast cancer susceptibility (BRCA) genes will sensitize cells to DNA damage- inducing therapeutic agents, including radiation therapy, cisplatin and PARP inhibitors. In the second aim, we will investigate the impact of ATM and PARP1 on Polθ is recruited to break sites and how the polymerase modulates damage sites to promote erroneous repair. In addition, we will pursue a proteomic-based approach to highlight the full spectrum of molecular players involved in alt-NHEJ. Ultimately, a full understanding of the mechanistic basis of alt-NHEJ will provide a better understanding of the source of genomic instability during the course of malignancy and guide more effective treatment strategies for the increasing number of patients with HR mutated tumors.

染色体重排是癌细胞的一个标志，并构成了一条主要途径影响肿瘤发生和进展的基因会发生突变。由双链断裂 (DSB) 修复缺陷引起的 DNA 有 3 个主要途径。哺乳动物细胞的修复——经过充分研究的同源重组（HR）和非同源重组末端连接 (NHEJ) 途径，以及特征较差但极易出错的 alt-NHEJ (替代-NHEJ)途径已被注意到与替代-NHEJ一致的遗传重排。在自发性肿瘤和治疗相关肿瘤中，我们将重点关注聚合酶θ。 (Polθ)，一种低保真酶，我们最近将其确定为介导 DSB 修复的关键因素鉴于 HR 缺陷的肿瘤“沉迷于”通过 alt-NHEJ 修复 DSB。途径，我们追求 Polθ 的诱变活性有助于建立基因组景观这有利于肿瘤的侵袭性行为。此外，我们预测删除肿瘤中的 Polθ。乳腺癌易感性 (BRCA) 基因突变会使细胞对 DNA 损伤变得敏感- 诱导治疗剂，包括放射治疗、顺铂和PARP抑制剂。第二个目标，我们将调查ATM和PARP1对Polθ被招募来破坏站点的影响以及聚合酶如何调节损伤位点以促进错误修复。追求基于蛋白质组学的方法来突出参与的全方位分子参与者最终，充分了解 alt-NHEJ 的机制基础将提供更好的结果。了解恶性肿瘤过程中基因组不稳定性的根源并指导更多针对越来越多的 HR 突变肿瘤患者的有效治疗策略。