Defining role of FANCA in genome instability

摘要 驱动癌症发展的最主要标志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所需的额外能力， 进展已知约400个基因发生突变并与癌症发展有关，其中大多数是直接的基因突变。 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要的。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，临床特征是骨髓 失败和易患癌症。这项提案的重点是FANCA，一个基因，是突变的约64%， 整个FA患者人群。在初步研究中，我们发现FANCA促进了易错DNA 驱动基因组不稳定性的修复;其表达在许多癌症类型中上调， 水平与乳腺癌进展密切相关，与癌症患者 生存有趣的是，FANCA募集到双链断裂和DNA损伤位点需要活性的 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在癌症中的高表达， 细胞促进易错修复、基因组不稳定性和细胞周期进程。描绘的作用 FANCA在基因组不稳定性和癌症发展中的作用，我们将使用生物化学定义的体外系统， 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，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;目的2是研究FANCA的作用。 FANCA在DSB介导的基因组不稳定性中的作用以及FANCA是如何调节的;目的3是确定FANCA在DSB介导的基因组不稳定性中的作用。 FANCA介导的基因组不稳定性与细胞周期进程之间的关系。完成本 该提案将确定FANCA在基因组不稳定性中的新作用。这项工作也将阐明的意义 FANCA作为一个独特的，理性驱动的癌症治疗靶点。这项提案的结果将扩大 FANCA表达升高和基因组不稳定的癌症患者的治疗策略。

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.