Defining role of FANCA in genome instability

FANCA 在基因组不稳定中的定义作用

基本信息

批准号：
10179004
负责人：
Yanbin Zhang
金额：
$ 48.61万
依托单位：
UNIVERSITY OF MIAMI SCHOOL OF MEDICINE
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：
2021-09-01 至 2024-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10179004
关键词：
Affect Automobile Driving BRCA1 gene Binding Binding Proteins Biochemical Biological Assay Breast Cancer Cell Breast Cancer cell line Cancer Patient Cell Aging Cell Cycle Cell Cycle Arrest Cell Cycle Progression Cells Cellular Assay Chromosome Fragile Sites Clinical Complement Coupled Crosslinker DNA Damage DNA Repair DNA annealing Data Defect Development Dose Etiology Fanconi Anemia Complementation Group A Protein Fanconi Anemia Complementation Group G Protein Fanconi Anemia pathway Fanconi&apos s Anemia G1 Arrest G2/M Arrest Genes Genetic Transcription Genomic Instability Goals Growth Hereditary Disease In Vitro Knock-out MDA MB 231 Malignant Neoplasms Mammary Neoplasms Maps Mediating Molecular Mus Mutate Mutation Nucleic Acid Binding Outcome Pancytopenia Pathway interactions Phosphorylation Predisposition Proteins RNA Reporter Research Resolution Role Site System Testing Transplantation Work Xenograft procedure base breast cancer progression cancer cell cancer therapy cancer type genome-wide human disease imaging system in vivo innovation knock-down live cell imaging malignant breast neoplasm mouse model mutant new therapeutic target novel overexpression patient population reconstitution recruit repaired senescence small molecule therapeutic target treatment strategy triple-negative invasive breast carcinoma tumor growth

项目摘要

Abstract One of the most predominant hallmarks driving cancer development is genome instability. It creates genome- wide diversity that enables cells to acquire additional capabilities required for cancer development and progression. Most of the ~400 genes known to be mutated and implicated in cancer development are a direct result of increased genome instability. Therefore, understanding the molecular mechanisms of genome instability in cancer cells is imperative for the development of novel treatment strategies. Fanconi Anemia (FA) is a hereditary disorder caused by mutations in at least 22 genes and clinically characterized by bone marrow failure and predisposition to cancer. This proposal focuses on FANCA, a gene that is mutated in ~64% of the entire FA patient population. During the preliminary studies, we found that FANCA promotes error-prone DNA repair that drives genome instability; its expression is upregulated in many cancer types, and the expression level is strongly associated with breast cancer progression and inversely correlates with cancer patient survival. Intriguingly, FANCA recruitment to double strand breaks and DNA damage sites requires active transcription in a KillerRed live cell analysis. More importantly, knockout of FANCA in a triple negative breast cancer cell MDA-MB-231 initiates cell cycle arrest and cellular senescence and abolishes breast cancer formation in mice. Based on these preliminary data, we hypothesize that high expression of FANCA in cancer cells promotes error-prone repair, genome instability, and cell cycle progression. To delineate the role of FANCA in genome instability and cancer development, we will use a biochemically defined in vitro system, a transcription-coupled DSB repair reporter system, a KillerRed live cell imaging system, a xenograft mouse model, and genome-wide instability analysis to accomplish three aims: Aim 1 is to determine the molecular mechanism of how FANCA contributes to R-loop-mediated genome instability; Aim 2 is to study the role of FANCA in DSB-mediated genome instability and how FANCA is regulated; Aim 3 is to determine the relationship between FANCA-mediated genome instability and cell cycle progression. Completion of this proposal will define a novel role for FANCA in genome instability. This work will also elucidate the significance of FANCA as a unique, rationale-driven target for cancer treatment. The outcome of this proposal will expand treatment strategies for cancer patients with elevated FANCA expression and genome instability.

抽象的推动癌症发展的最主要标志之一是基因组不稳定性。它创建基因组 - 使细胞能够获得癌症发展所需的其他功能的广泛多样性和进展。已知突变和与癌症发展有关的〜400个基因中的大多数是直接的基因组不稳定性增加的结果。因此，了解基因组的分子机制癌细胞中的不稳定性对于发展新型治疗策略至关重要。 Fanconi贫血（FA）是由至少22个基因突变引起的遗传性疾病，并以骨髓为特征癌症的失败和易感性。该提案的重点是Fanca，该基因在约64％的基因中突变整个FA患者人数。在初步研究期间，我们发现FANCA促进了容易出错的DNA 维修驱动基因组不稳定性的修复；它的表达在许多癌症类型中被上调，表达水平与乳腺癌的进展密切相关，并与癌症患者成反关系生存。有趣的是，FANCA招募进行双链断裂和DNA损伤部位需要活动在杀伤活细胞分析中转录。更重要的是，在三重负乳房中敲除芬卡癌细胞MDA-MB-231启动细胞周期停滞和细胞衰老，并废除乳腺癌小鼠的形成。基于这些初步数据，我们假设癌症在癌症中的高表达细胞促进容易产生错误的修复，基因组不稳定性和细胞周期进程。描述基因组不稳定性和癌症发展中的粉丝，我们将使用体外系统定义的生化定义转录耦合DSB维修报告机系统，一种Killerred活细胞成像系统，异种移植小鼠模型和全基因组不稳定性分析以实现三个目标：目标1是确定分子 Fanca如何促进R环介导的基因组不稳定性的机制；目标2是研究 DSB介导的基因组不稳定性以及如何调节Fanca的Fanca；目标3是确定 FANCA介导的基因组不稳定性与细胞周期进程之间的关系。完成此操作提案将定义FANCA在基因组不稳定性中的新作用。这项工作还将阐明重要性 FANCA是癌症治疗的独特，理由驱动的靶标。该提议的结果将扩大 Fanca表达和基因组不稳定性升高的癌症患者的治疗策略。