Therapeutic potential of FANCM for BRCA1-linked cancer

FANCM 对 BRCA1 相关癌症的治疗潜力

• 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
• 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' 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

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

项目摘要/摘要 乳腺癌和卵巢癌是全球女性最常见的癌症之一。PARP抑制剂具有 通过合成的致死性机制，显示了对BRCA连接癌症治疗的潜力 人力资源缺陷。但是，肿瘤通常会对这些药物产生抗性。因此，有一个紧迫的 需要寻找新的针对性癌症的针对性治疗方法。基因组不稳定性是癌细胞的标志 和肿瘤发生的潜在来源。基因组不稳定性的主要原因是复制叉在站点停滞不前 DNA损伤或异常DNA结构。哺乳动物失速的叉子修复的局限性是 用分子细节分析此过程的工具的死亡。 Scully Lab通过 适应大肠杆菌/ter复制叉屏障（RFB）以诱导特定地点复制叉 储存在哺乳动物染色体上。 缺乏BRCA1的原代细胞中的串联重复（TDS）是由TUS/TER块专门诱导的，但不是 通过常规的双股断裂（DSB），表明停滞的叉子响应的特异性。有趣的是， 缺乏BRCA1的乳腺癌和卵巢癌类似地获得了大量的小（〜10 kb）TD，我们 已称为“第1组” TD。那就是TUS/TER系统概括了BRCA1特定的调节 在人乳腺癌和卵巢癌中观察到的1组TD形成。我发现停滞的叉子电机 蛋白质-Fancm（Fanconi贫血[FA]组M Gene的产物）与BRCA1协同作用 抑制TU/TER诱导的TD。此外，我发现了BRCA1之间的一种新型的合成致命相互作用 小鼠胚胎（ES）细胞以及乳腺癌和卵巢癌细胞中的粉丝流失。这些发现 表明FANCM可能是BRCA1连接乳腺癌和卵巢癌的有前途的治疗靶标。 我在此提案中的目标是描述癌细胞中新型的粉丝 - BRCA1合成致命相互作用 并确定合成致死性的机制（AIM1）。此外，我将探索染色质环境 和停滞的叉子的蛋白质动力学，并将研究这些过程中的变化如何有助于 FANCM-BRCA1合成致命相互作用（AIM2）。我观察到Fancm及其下游的上任角色 TARGET FANCD2在TUS/TER中促进无错误修复和抑制易于错误的修复。 FANCM和FANCD2处于维修途径的选择中，叉子会增加他们可能共享类似的可能性 与BRCA1的遗传相互作用。将确定FANCD2的单个域在维修途径中的功能 停滞叉子的选择及其与BRCA1的遗传相互作用（AIM 3）。这种整体方法将提供完整的 FANCM-BRCA1合成致命相互作用的机理图片，可能在FANCD2中识别一个新的 癌症治疗的合成致命靶标。 我