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

描述（由申请人提供）：Poly（ADP-ribosyl）化是一种翻译后修饰，在细胞死亡和DNA修复的调控中起着核心作用。PAR作为多聚（adp -核糖）聚合酶（PARP）和多聚（adp -核糖）糖水解酶（PARG）的替代标记物，这两种酶正被积极地作为治疗靶点，一些PARP抑制剂已经在临床试验中。了解PARP调控和PARP/PARG药物发现的核心是提供敏感、简单和低成本的检测方法，这些方法目前尚无。因此，本1期申请旨在利用工程分裂蛋白报告体开发PAR的均质分析。提出的新的药效学和细胞分析将有助于改进治疗方案，并加速学术界和工业界对PARP和PARG抑制剂的开发。

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.