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）：在乳腺癌敏感性基因BRCA1中携带突变的个体倾向于早期发作乳腺癌和卵巢癌，这是西方社会中一些最常见的恶性肿瘤。 BRCA1中的这种突变几乎是所有遗传性乳腺癌和卵巢癌的家庭，以及大约一半的乳腺癌家庭。从BR​​CA1载体中影响野生型BRCA1等位基因的杂合性丧失（LOH）的检测意味着BRCA1是肿瘤抑制器。 BRCA1参与乳腺癌很复杂。迄今为止，已经确定了100多种自然存在的BRCA1种系突变。对BRCA1的研究对乳腺癌研究具有重要意义，并试图阐明其生化功能的尝试包括确定其蛋白质伴侣，例如BAP1。 BAP1是去泛素化酶（DUB）的UCH家族的成员。这些是蛋白酶，可以逆转泛素偶联到蛋白质的蛋白质，以蛋白酶体或重新定位响应泛素化。泛素的结合已被证明在控制许多调节途径（包括：细胞周期调节，染色质结构，DNA修复和基因组稳定性，转录，病毒发病机理，免疫反应和蛋白质质量控​​制。在至少某些病理状况下，去泛素化酶可能是有用的药物靶标。除神经元UCH-L1外，没有配音是系统屏幕的目标。我们已经开发了以负担得起的成本生产大量通用配音底物的方法，并选择将精力集中在与BRCA1肿瘤抑制剂相关的配音上。我们已经开始优化筛选条件，并表明该测定法适应384井格式。我们将安装一个常规的高吞吐量药物屏幕，其中可用的NIH化合物库，以识别DUB作用的抑制剂和激活剂。该屏幕将导致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。