HTS for Inhibitors of BAP1,BRCA:Deubiquinating(RMI)

BAP1、BRCA 抑制剂的 HTS：去泛素化 (RMI)

• 批准号: 7058567
• 负责人: KEITH D WILKINSON
• 金额: $ 0.46万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-15 至 2006-09-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7058567
• 关键词: brca gene; breast neoplasms; chemical registry /resource; drug discovery /isolation; drug screening /evaluation; endopeptidases; enzyme inhibitors; enzyme substrate; gene mutation; genetic susceptibility; high throughput technology; neoplasm /cancer genetics; ovary neoplasms; tissue /cell culture; ubiquitin

DESCRIPTION (provided by applicant): Individuals who carry mutations in the breast cancer susceptibility gene, BRCA1, are predisposed to early onset breast and ovarian cancer, some of the most common malignancies in Western societies. Such mutations in BRCA1 account for almost all families with inherited breast and ovarian cancer and for approximately half of families with breast cancer only. The detection of loss-of-heterozygosity (LOH) affecting the wild-type BRCA1 allele in tumors from BRCA1 carriers implies that BRCA1 is a tumor suppressor. The involvement of BRCA1 in breast cancer is complex; to date, more than 100 unique, naturally occurring BRCA1 germline mutations have been identified. The study of BRCA1 has important implications for breast cancer research and attempts to elucidate its biochemical function have included identifying its protein partners, such as BAP1. BAP1 is a member of the UCH family of deubiquitinating enzymes (DUB). These are proteases that reverse the conjugation of ubiquitin to proteins targeted for degradation by the proteasome or relocalization in response to ubiquitination. The conjugation of ubiquitin has been shown to be important in control of many regulatory pathways including; cell cycle regulation, chromatin structure, DNA repair and genome stability, transcription, viral pathogenesis, immune response, and protein quality control. Deubiquitinating enzymes are likely to be useful drug targets in at least some pathological conditions. With the exception of neuronal UCH-L1, no DUB has been the target of a systematic screen. We have developed the means to produce significant amounts of a generic DUB substrate at affordable costs and have chosen to focus our efforts first on a DUB associated with the BRCA1 tumor suppressor. We have begun to optimize the screening conditions and have shown that the assay is adaptable to the 384 well format. We will mount a conventional high throughput drug screen of available NIH compound libraries to identify inhibitors and activators of DUB action. This screen will result in useful molecular probes of BAP1 function and help to clarify the role of BAP1 in BRCA1 mediated events.

描述（由申请人提供）：携带乳腺癌易感基因 BRCA1 突变的个体易患早发性乳腺癌和卵巢癌，这是西方社会最常见的恶性肿瘤之一。 BRCA1 的此类突变几乎导致所有患有遗传性乳腺癌和卵巢癌的家庭，以及大约一半仅患有乳腺癌的家庭。在 BRCA1 携带者的肿瘤中检测到影响野生型 BRCA1 等位基因的杂合性丢失 (LOH) 意味着 BRCA1 是一种肿瘤抑制因子。 BRCA1 与乳腺癌的关系很复杂；迄今为止，已鉴定出 100 多种独特的、自然发生的 BRCA1 种系突变。 BRCA1 的研究对乳腺癌研究具有重要意义，阐明其生化功能的尝试包括鉴定其蛋白质伙伴，例如 BAP1。 BAP1 是 UCH 去泛素化酶 (DUB) 家族的成员。这些蛋白酶可逆转泛素与蛋白质的缀合，从而被蛋白酶体降解或响应泛素化而重新定位。泛素的结合已被证明对于控制许多调控途径非常重要，包括：细胞周期调控、染色质结构、DNA 修复和基因组稳定性、转录、病毒发病机制、免疫反应和蛋白质质量控​​制。至少在某些病理条件下，去泛素化酶可能是有用的药物靶点。除了神经元 UCH-L1 之外，没有 DUB 成为系统筛选的目标。我们已经开发出以可承受的成本生产大量通用 DUB 底物的方法，并选择首先将我们的努力集中在与 BRCA1 肿瘤抑制因子相关的 DUB 上。我们已经开始优化筛选条件，并表明该测定适用于 384 孔格式。我们将对可用的 NIH 化合物库进行常规高通量药物筛选，以鉴定 DUB 作用的抑制剂和激活剂。该筛选将产生 BAP1 功能的有用分子探针，并有助于阐明 BAP1 在 BRCA1 介导的事件中的作用。