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

描述（由申请人提供）：携带乳腺癌易感基因BRCA 1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。BRCA 1中的这种突变几乎可以解释所有遗传性乳腺癌和卵巢癌家族，以及大约一半仅患有乳腺癌的家族。在BRCA 1携带者的肿瘤中检测到影响野生型BRCA 1等位基因的杂合性缺失（洛），这意味着BRCA 1是一种肿瘤抑制因子。BRCA 1在乳腺癌中的参与是复杂的;迄今为止，已经确定了100多个独特的、自然发生的BRCA 1种系突变。BRCA 1的研究对乳腺癌研究具有重要意义，试图阐明其生化功能包括鉴定其蛋白质伴侣，如BAP 1。BAP 1是去泛素化酶（DUB）UCH家族的成员。这些蛋白酶是逆转泛素与蛋白质的缀合的蛋白酶，所述蛋白质被蛋白酶体降解或响应于泛素化而重新定位。泛素的缀合已被证明在控制许多调节途径中是重要的，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、病毒发病机制、免疫应答和蛋白质质量控制。去泛素化酶可能是至少在某些病理条件下有用的药物靶标。除神经元UCH-L1外，没有DUB是系统筛选的目标。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，并选择将我们的努力首先集中在与BRCA 1肿瘤抑制因子相关的DUB上。我们已经开始优化筛选条件，并已表明该测定适用于384孔格式。我们将对现有的NIH化合物库进行常规的高通量药物筛选，以鉴定DUB作用的抑制剂和激活剂。该筛选将产生有用的BAP 1功能的分子探针，并有助于阐明BAP 1在BRCA 1介导的事件中的作用。

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.