High Throughput Screening Assay for IP7K inositol pyrophosphate kinases

 描述(由申请人提供)：这项拟议的合作是北卡罗来纳大学教堂山分校的综合化学生物学和药物发现中心(CICBDD)和三角研究园区NIEHS的Stephen Shears博士之间的合作。我们建议开发一种方法，应用于高通量筛选，以识别铅化学“探针”，以抑制一对功能专门化的小分子激酶，称为IP7K(又名。PPIP5Ks)。这些细胞信号转导酶(IP7K1/PPIP5K1和IP7K2/PPIP5K2)合成二磷酸肌醇磷酸：二磷肌醇五磷酸脂，1-IP7，和双二磷肌醇四氢磷酸盐，IP8。这些由IP7Ks催化的反应通过激活蛋白激酶AKT来促进细胞增殖。此外，控制促炎干扰素转录的分子通路在病毒感染期间被IP7K激活，这些相同的通路激活干扰素转录，以响应代谢性炎症，如糖尿病和肥胖症的特征。因此，IP7K是新的抗癌和抗炎治疗的潜在药理靶点。改变IP7K活性的探针的发展也为研究人员提供了识别和研究IP7K的额外功能的机会。因此，这项应用的重点是开发和验证适用于高通量筛选的分析方法，并伴随着正交分析和细胞内分析，以筛选化学文库，目的是发现新的细胞修饰的IP7K活性。

 DESCRIPTION (provided by applicant): This proposed collaboration is between the Center for Integrative Chemical Biology and Drug Discovery (CICBDD) at the University of North Carolina, Chapel Hill, and Dr. Stephen Shears at NIEHS, Research Triangle Park. We propose to develop the methodology that can applied to a high throughput screen to identify lead chemical "probes" to inhibit a functionally-specialized pair of small molecule kinases known as IP7Ks (a.k.a. PPIP5Ks). These cell-signaling enzymes (IP7K1/PPIP5K1 and IP7K2/PPIP5K2), synthesize diphosphorylated inositol phosphates: the diphosphoinositol pentakisphosphate, 1-IP7, and bis-diphosphoinositol tetrakisphosphate, IP8. These reactions catalyzed by IP7Ks promote cell proliferation through activation of the protein kinase, AKT. Moreover, molecular pathways controlling pro-inflammatory interferon transcription are activated by IP7Ks during viral infection These same pathways activate interferon transcription in response to metabolic inflammation, such as that which characterizes diabetes and obesity. Thus, IP7Ks are potential pharmacological targets for new anti-cancer and anti- inflammatory therapies. The development of probes that alter the activities of IP7Ks also offers opportunities for researchers to identify and study additional functions of the IP7Ks. Thus, the focus of this application is to develop and validate assay methods applicable to high throughput screening, with accompanying orthogonal and in-cell assays, to screen chemical libraries with the aim of discovering novel, cell- permeant modifiers of IP7K activity.