Therapeutic potential of FANCM for BRCA1-linked cancer

FANCM 对 BRCA1 相关癌症的治疗潜力

基本信息

批准号：
10446024
负责人：
Arvind Panday
金额：
$ 9.95万
依托单位：
BETH ISRAEL DEACONESS MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-04-01 至 2024-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10446024
关键词：
BRCA1 gene Biochemical CRISPR/Cas technology Cancer cell line Cells Chromatin Chromosome Structures Chromosomes Cytogenetics DNA DNA Damage DNA Structure DNA replication fork Defect Environment Escherichia coli Event FANCD2 protein Fanconi&apos s Anemia Gene Conversion Genes Genetic Genetic study Genomic Instability Goals Hereditary Breast and Ovarian Cancer Syndrome Histones Human Individual Knock-out Link Malignant Neoplasms Malignant neoplasm of ovary Mammalian Cell Mammalian Chromosomes Mediating Molecular Motor Mus Mutation Nucleosomes Outcome Pathway interactions Pharmaceutical Preparations Phase Phenocopy Phenotype Play Point Mutation Problem Solving Process Protein Dynamics Proteins Regulation Reporter Resistance development Role Site Small Interfering RNA Source Specificity Susceptibility Gene System Techniques Therapeutic Woman Work cancer cell cancer genome cancer predisposition cancer therapy chromatin remodeling embryonic stem cell genome-wide holistic approach homologous recombination inhibitor knock-down malignant breast neoplasm mutant novel novel therapeutics prevent recruit repaired response success targeted cancer therapy therapeutic target tool translocase tumor tumorigenesis

项目摘要

Project Summary/Abstract Breast and ovarian cancer are among the most common cancers in women worldwide.PARP inhibitors have shown potential for the treatment of BRCA-linked cancer, via a synthetic lethality mechanism that exploits the HR defect. However, tumors often develop resistance to these and other drugs. Therefore, there is a pressing need to find new, targeted treatments for BRCA-linked cancer. Genomic instability is a hallmark of cancer cells and a potential source of tumorigenesis. A major cause of genomic instability is replication fork stalling at sites of DNA damage or abnormal DNA structure. A limitation in the study of mammalian stalled fork repair has been a dearth of tools with which to analyze this process in molecular detail. The Scully lab solved this problem by adapting the Escherichia coli Tus/Ter replication fork barrier (RFB) to induce site-specific replication fork stalling on a mammalian chromosome. Tandem duplications (TDs) in primary cells lacking BRCA1 are induced specifically by a Tus/Ter block but not by a conventional double strand break (DSB), indicating specificity for the stalled fork response. Intriguingly, breast and ovarian cancers lacking BRCA1 similarly acquire large numbers of small (~10 kb) TDs, which we have termed “Group 1” TD. Thus, the Tus/Ter system recapitulates the BRCA1-specific regulation of Group 1 TD formation observed in human breast and ovarian cancer. I found that the stalled fork motor protein—FANCM (product of the Fanconi anemia [FA] group M gene) acts synergistically with BRCA1 to suppress Tus/Ter-induced TDs. Further, I discovered a novel synthetic lethal interaction between Brca1 and Fancm loss in mouse embryonic stem (ES) cells and in breast and ovarian cancer cells. These findings suggest that FANCM may be a promising therapeutic target in BRCA1-linked breast and ovarian cancer. My goals in this proposal are to delineate the novel FANCM-BRCA1 synthetic lethal interaction in cancer cells and to determine the mechanism of synthetic lethality (Aim1). Further, I will explore the chromatin environment and protein dynamics at the stalled fork and will study how alterations in these processes contribute to the FANCM-BRCA1 synthetic lethal interaction (Aim2). I observe an epistatic role of Fancm and its downstream target Fancd2 at Tus/Ter in promoting error free repair and suppressing error-prone repair.This critical role of Fancm and Fancd2 in repair pathway choice at stalled forks raises the possibility that they might share similar genetic interactions with Brca1. will identify how individual domains of FANCD2 function in repair pathway choice at stalled forks and their genetic interaction with Brca1 (Aim 3). This holistic approach will provide a full picture of the mechanism of FANCM-BRCA1 synthetic lethal interactions and might identify in FANCD2 a new synthetic lethal target for cancer therapy. I

项目概要/摘要乳腺癌和卵巢癌是全球女性最常见的癌症之一。PARP 抑制剂已显示出通过利用 BRCA 相关癌症的合成致死机制治疗 BRCA 相关癌症的潜力然而，肿瘤常常对这些药物和其他药物产生耐药性，因此存在紧迫的问题。需要寻找新的、针对 BRCA 相关癌症的靶向治疗方法。基因组不稳定是癌细胞的一个标志。肿瘤发生的一个潜在来源是基因组不稳定的一个主要原因是复制叉在位点上停滞。 DNA 损伤或异常 DNA 结构的研究受到限制。由于缺乏对这一过程进行分子详细分析的工具，斯卡利实验室通过以下方法解决了这个问题。调整大肠杆菌 Tus/Ter 复制叉屏障 (RFB) 以诱导位点特异性复制叉停顿在哺乳动物染色体上。缺乏 BRCA1 的原代细胞中的串联重复 (TD) 是由 Tus/Ter 阻断特异性诱导的，但不是通过传统的双链断裂（DSB），表明停滞叉响应的特异性。缺乏 BRCA1 的乳腺癌和卵巢癌同样会获得大量小 (~10 kb) TD，我们将其被称为“第 1 组”TD 因此，Tus/Ter 系统概括了 BRCA1 特异性调节。在人类乳腺癌和卵巢癌中观察到第 1 组 TD 形成，我发现叉形电机失速。蛋白质 — FANCM（范可尼贫血 [FA] M 组基因的产物）与 BRCA1 协同作用，抑制 Tus/Ter 诱导的 TDs 此外，我发现了 Brca1 之间的一种新的合成致死相互作用。小鼠胚胎干 (ES) 细胞以及乳腺癌和卵巢癌细胞中 Fancm 损失。表明 FANCM 可能是 BRCA1 相关乳腺癌和卵巢癌的一个有前途的治疗靶点。我在本提案中的目标是描述癌细胞中新型 FANCM-BRCA1 合成致死相互作用并确定合成致死的机制（目标1）。此外，我将探索染色质环境。和停滞叉处的蛋白质动力学，并将研究这些过程的改变如何促进 FANCM-BRCA1 合成致死相互作用 (Aim2) 我观察到 Fancm 及其下游的上位作用。 Tus/Ter 上的目标 Fancd2 在促进无错修复和抑制易错修复方面发挥着关键作用 Fancm 和 Fancd2 在停滞分叉处的修复途径选择中提出了它们可能具有相似的可能性与 Brca1 的遗传相互作用将确定 FANCD2 的各个域如何在修复途径中发挥作用。停滞叉的选择及其与 Brca1 的遗传相互作用（目标 3）。 FANCM-BRCA1 合成致死相互作用的机制图，并可能在 FANCD2 中识别出新的癌症治疗的合成致死靶标。我