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：
10471217
负责人：
Yanbin Zhang
金额：
$ 47.27万
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/10471217
关键词：
Affect Automobile Driving BRCA1 gene Binding Binding Proteins Biochemical Biological Assay Bone marrow failure 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 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.

抽象的驱动癌症发展的最显着特征之一是基因组不稳定性。它创造了基因组- 广泛的多样性使细胞能够获得癌症发展和治疗所需的额外能力进展。已知大约 400 个突变并与癌症发展有关的基因中的大多数都是直接基因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的不稳定性对于开发新的治疗策略至关重要。范可尼贫血 (FA) 是一种由至少 22 个基因突变引起的遗传性疾病，临床特征为骨髓失败和患癌症的倾向。该提案重点关注 FANCA，该基因在约 64% 的人中发生突变整个 FA 患者群体。在初步研究中，我们发现 FANCA 会促进容易出错的 DNA 导致基因组不稳定的修复；它的表达在许多癌症类型中上调，并且表达水平与乳腺癌进展密切相关，并与癌症患者呈负相关生存。有趣的是，FANCA 招募双链断裂和 DNA 损伤位点需要主动 KillerRed 活细胞分析中的转录。更重要的是，在三阴性乳腺癌中敲除 FANCA 癌细胞 MDA-MB-231 启动细胞周期停滞和细胞衰老并消除乳腺癌在小鼠中形成。基于这些初步数据，我们假设 FANCA 在癌症中高表达细胞促进易错修复、基因组不稳定和细胞周期进展。界定角色 FANCA 在基因组不稳定性和癌症发展方面，我们将使用生化定义的体外系统，转录耦合 DSB 修复报告系统、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来实现三个目标：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中的 FANCA 以及 FANCA 的调节方式；目标 3 是确定 FANCA介导的基因组不稳定性与细胞周期进展之间的关系。完成此该提案将定义 FANCA 在基因组不稳定性方面的新作用。这项工作也将阐明其意义 FANCA 是一个独特的、理性驱动的癌症治疗目标。该提案的成果将扩大 FANCA 表达升高和基因组不稳定的癌症患者的治疗策略。