ACTIVITY-BASED KINASE DISCOVERY IN SMALL CELL LUNG CANCER

描述（由申请人提供）：小细胞肺癌（SCLC）是一种尚未得到充分研究的癌症，目前还没有分子靶向方法显示出其用途。在这里，我们建议使用化学生物学平台，基于活性的蛋白质分析（ABPP）来研究小细胞肺癌（SCLC）蛋白质组。 ABPP 使用针对酶活性位点的化学探针来询问生物样品中酶的功能状态。使用 ABPP 研究了许多酶家族，包括丝氨酸水解酶、激酶、磷酸酶和金属蛋白酶。我们建议使用 ABPP 根据全局蛋白激酶分析更好地对 SCLC 进行分类和亚组，这可以指导未来开发靶向治疗的尝试。该项目的假设是，基于激酶组的 ABPP 可以检测 SCLC 中的功能激酶，并通过以下方式指导未来的临床管理：(i) 识别作为 SCLC 生长和生存的潜在驱动因素的活性激酶和通路，(ii) 可以导致开发对 SCLC 不同亚组进行分类的生物标志物，以及 (iii) 可以提供可进一步研究 SCLC 有效性的联合治疗方法。蛋白质组学分析提供了除基因测序或基因表达谱之外的 SCLC 生物学的额外视图，因此与检查 SCLC 生物学的其他重要工作相补充。为了实现项目目标，我们提出了四个目标。目标 1 将开发利用质谱分析表征激酶组和其他 ATP 结合蛋白的定量方法。目标 2 将使用 ABPP 表征 SCLC 肿瘤和细胞系中的激酶和其他 ATP 结合蛋白。目标 3 将使用蛋白质印迹和反相蛋白阵列验证 ABPP 的结果。目标 4 将使用 RNAi 和小分子抑制剂探讨 ABPP 识别的蛋白激酶的功能意义，以更好地指导生物标志物选择。

DESCRIPTION (provided by applicant): Small cell lung cancer (SCLC) is an understudied cancer for which no molecularly targeted approaches have shown use. Here we propose to use a chemical biology platform, activity-based protein profiling (ABPP), to study the small cell lung cancer (SCLC) proteome. ABPP uses chemical probes that are directed against the active sites of enzymes to interrogate the functional state of enzymes in biological samples. Using ABPP, a number of enzyme families have been studied, including serine hydrolases, kinases, phosphatases, and metalloproteinases. We propose to use ABPP to better classify and subgroup SCLC based on global protein kinase profiling that can guide future attempts towards developing targeted therapy. The hypothesis for this project is that kinome-based ABPP can detect functional kinases in SCLC and guide future clinical management by (i) identifying active kinases and pathways that are potential drivers of SCLC growth and survival, (ii) can lead to development of biomarkers that classify different subgroups of SCLC, and (iii) can inform about combination therapy approaches that can be further studied for effectiveness in SCLC. Proteomic profiling offers additional views of SCLC biology beyond sequencing of genes or gene expression profiles and thus is complementary to other important work examining SCLC biology. To enable the project goals, we have proposed four aims. Aim 1 will develop quantitative methods to characterize kinome and other ATP-binding proteins using mass spectrometry. Aim 2 will characterize kinases and other ATP-binding proteins in SCLC tumors and cell lines using ABPP. Aim 3 will validate findings of ABPP findings using western blotting and Reverse Phase Protein Arrays. Aim 4 will interrogate the functional significance of ABPP identified proteins kinases using RNAi and small molecule inhibitors to better guide biomarker selection.