Chemical Probes That Modulate Phosphatidylinositol-5-Phosphate 4-Kinase Activity

描述（由申请人提供）：本提案旨在鉴定和优化人PI 5 P4 K、磷脂酰肌醇-5-磷酸4-激酶（II型PIPK）（调节AKT信号传导和肿瘤细胞生长的激酶家族）的特异性调节剂。将追求以下具体目标：（1）使用针对含有约400，000个小分子的MLSMR的定量高通量筛选方法鉴定人PI 5 P4 K的抑制剂。(2)在一组二级试验中验证已鉴别化合物的效价，该试验包括选择性试验，以鉴别不影响PI 4P 5 K、磷脂酰肌醇-4-磷酸5-激酶（I型PIPK）和PI 3 P5 K、磷脂酰肌醇-3-磷酸5-激酶（III型PIPK /Fab 1/PIKfyve）的化合物。我们将使用PI 5 P4 K敲除小鼠胚胎成纤维细胞（MEF）和敲减细胞作为对照，测试这些PI 5 P4 K探针对细胞PI 5 P积累和细胞增殖的影响和特异性。(3)除了这项提议的授权期限外，还将检查最有效的PI 5 P4 K探针在小鼠癌症模型中减少肿瘤生长的能力。

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.