A Poly(ADP-Ribose) Detection Assay Enabling Drug Discovery and Development

描述（由申请人提供）：聚（ADP-核糖基）化是一种翻译后修饰，在细胞死亡和DNA修复的调节中起着核心作用。PAR作为聚（ADP-核糖）聚合酶（PARP）和聚（ADP-核糖）糖水解酶（PARG）的替代标志物，这些标志物正被积极用作临床试验中几种PARP抑制剂的治疗靶点。理解PAR调节和PARP/PARG药物发现的核心是敏感、简单和低成本的检测方法的可用性，这些方法目前尚不可用。因此，该第1阶段申请寻求开发利用工程化分裂蛋白报告基因的PAR的均相测定。提出的新的药效学和细胞测定将有助于推进治疗方案，并加速学术界和工业界对PARP和PARG抑制剂的开发。 公共卫生相关性：聚腺苷二磷酸核糖基化是蛋白质的翻译后修饰，在环境和代谢应激引起的单链断裂修复中起着重要作用。细胞中的聚（ADP-核糖）水平由PARP和PARG的双重作用维持，PARP合成PAR; PARG降解细胞中的PAR。因此，PAR可用于报告这两种酶的活性。本申请的目的是开发用于药物发现的低成本、灵敏、基于发光的聚（ADP-核糖）检测分析。

DESCRIPTION (provided by applicant): Poly(ADP-ribosyl)ation is a post-translational modification that plays a central role in the regulation of cell death and DNA repair. PAR serves as a surrogate marker for poly(ADP-ribose) polymerase (PARP) and poly(ADP-ribose) glycohydrolase (PARG), which are being actively pursued as therapeutic targets with several PARP inhibitors already in clinical trials. Central to understanding PAR regulation and PARP/PARG drug discovery would be the availability of sensitive, simple and low-cost assays, which are currently unavailable. Thus this Phase 1 application seeks to develop homogeneous assays for PAR utilizing engineered split-protein reporters. The new pharmacodynamic and cellular assays that are proposed will help advance treatment regimens as well as accelerate the development of PARP and PARG inhibitors in both academia and industry. PUBLIC HEALTH RELEVANCE: Poly (ADP-ribosylation), a post-translational modification of proteins, plays a central role in the repair of single strand breaks caused by environmental and metabolic stress. Poly(ADP-ribose) levels in cells are maintained by the dual action of PARP, which synthesizes PAR; and PARG, which degrades PAR in cells. Hence PAR can be used to report on the activity of both these enzymes. The purpose of our application is to develop low-cost, sensitive, luminescence based poly(ADP-ribose) detection assays for drug discovery.