Defining role of FANCA in genome instability

摘要 推动癌症发展的最主要特征之一是基因组不稳定。它创造了基因组- 广泛的多样性，使细胞能够获得癌症发展所需的额外能力 进步。已知的与癌症发生有关的~400个基因中的大多数都是直接的 基因组不稳定性增加的结果。因此，了解基因组的分子机制 癌细胞的不稳定性对于开发新的治疗策略是必不可少的。范可尼贫血(FA) 是一种遗传性疾病，由至少22个基因突变引起，临床特征为骨髓 失败和易患癌症。这项建议的重点是FANCA，一种在~中突变的基因 整个FA患者群体。在初步研究中，我们发现FANCA促进了容易出错的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 水平与乳腺癌的进展密切相关，与癌症患者呈负相关 生死存亡。有趣的是，FANCA招募双链断裂和DNA损伤部位需要积极 KillerRed活细胞分析中的转录。更重要的是，三重阴性乳房中的FANCA基因敲除 癌细胞MDA-MB-231启动细胞周期停滞和细胞衰老并消灭乳腺癌 在小鼠体内形成。基于这些初步数据，我们假设FANCA促进了基因组的不稳定性， 调节细胞周期进程，并作为癌症干预的脆弱性。描绘…的角色 在基因组不稳定和癌症发展方面，我们将使用一个生物化学定义的体外系统，一个 转录偶联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实现了三个目标：目标1是确定分子 FANCA在R环修复中的作用机制；目的2是研究FANCA在基因组中的作用 不稳定以及FANCA是如何调控的；目标3是确定FANCA在细胞周期进程中的作用和 肿瘤生长。这项提议的完成将定义FANCA在基因组不稳定中的新角色。这部作品 还将阐明FANCA作为一种独特的、理性驱动的癌症治疗靶点的意义。这个 这项提案的结果将扩大FANCA表达升高的癌症患者的治疗策略 和基因组的不稳定性。

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 FANCA promotes genome instability, regulates cell cycle progression, and serves as a vulnerability for cancer intervention. 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 repair; Aim 2 is to study the role of FANCA in genome instability and how FANCA is regulated; Aim 3 is to determine the role of FANCA in cell cycle progression and tumor growth. 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.