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

描述(由申请人提供)：聚(ADP-核糖基)修饰是一种翻译后修饰，在调节细胞死亡和DNA修复中发挥核心作用。PAR是聚ADP-核糖聚合酶(PARP)和聚(ADP-核糖)糖水解酶(PARG)的替代标记物，目前已有几种PARP抑制剂作为治疗靶点。了解PAR法规和PARP/PARG药物发现的核心是获得敏感、简单和低成本的分析，而这些目前还无法获得。因此，这一阶段的应用试图利用工程分离蛋白报告开发PAR的均质分析方法。所提出的新的药效学和细胞分析方法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：Poly(ADP-核糖化)是一种蛋白质的翻译后修饰，在修复环境和代谢应激引起的单链断裂方面发挥着核心作用。细胞中的多聚(ADP-核糖)水平是由合成PAR的PARP和降解细胞中PAR的PARG的双重作用维持的。因此，PAR可以用来报告这两种酶的活性。我们应用的目的是开发低成本、灵敏、基于发光的聚(ADP-核糖)检测方法用于药物发现。

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.