Chemical probes that modulate a stress pathway phosphatidylinositol 5-phosphate 4

描述（由申请人提供）：该提案旨在鉴定和优化人类PI5P4K，磷脂酰肌醇- 5-磷酸4激酶（II型PIPK）的特异性调节剂，这是一个调节AKT信号传导和肿瘤细胞生长的激酶家族。以下具体目标将被追求：(1)使用定量高通量筛选方法针对含有约400,000个小分子的MLSMR鉴定人类PI5P4K抑制剂。(2)在一组二级分析中验证鉴定的化合物的效价，二级分析包括选择性分析，以鉴定不影响PI4P5K，磷脂酰肌醇- 4-磷酸5激酶（I型PIPK）和PI3P5K，磷脂酰肌醇-3-磷酸5激酶（III型PIPK /Fab1/PIKfyve）的化合物。我们将使用PI5P4K敲除小鼠胚胎成纤维细胞（mef）和敲除细胞作为对照，测试这些PI5P4K探针对细胞PI5P积累和细胞增殖的影响和特异性。(3)在本提案的授权期限之外，研究最有效的PI5P4K探针在小鼠癌症模型中降低肿瘤生长的能力。

DESCRIPTION (provided by applicant): This proposal aims to identify and optimize specific modulators of human PI5P4K, phosphatidylinositol- 5-phosphate 4-kinase (Type II PIPK), a family of kinases that regulate AKT signaling and tumor cell growth. The following specific aims will be pursued: (1) to identify inhibitors of human PI5P4K using a quantitative high-throughput screening approach against the MLSMR containing ~400,000 small molecules. (2) To validate the potency of the identified compounds in a panel of secondary assays, which consist of selectivity assays to identify those compounds that do not affect PI4P5K, phosphatidylinositol- 4-phosphate 5-kinase (Type I PIPK), and PI3P5K, the phosphatidylinositol-3-phosphate 5-kinase (Type III PIPK /Fab1/PIKfyve). We will test the impact and specificity of these PI5P4K probes on cellular PI5P accumulation and cell proliferation using PI5P4K knockout mouse embryonic fibroblasts (MEFs) and knockdown cells as controls. (3) Beyond the granting term for this proposal, to examine the most potent PI5P4K probes for their ability to decrease tumor growth in mouse models of cancer.