ACTIVITY-BASED KINASE DISCOVERY IN SMALL CELL LUNG CANCER

描述（由申请人提供）：小细胞肺癌（SCLC）是一种研究不足的癌症，没有分子靶向方法显示出用途。在这里，我们建议使用化学生物学平台，基于活性的蛋白质谱（ABPP），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，已经研究了许多酶家族，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更好地分类和亚组SCLC的基础上，全球蛋白激酶谱，可以指导未来的尝试开发靶向治疗。该项目的假设是，基于激酶组的ABPP可以检测SCLC中的功能性激酶，并通过以下方式指导未来的临床管理：（i）识别作为SCLC生长和生存潜在驱动因素的活性激酶和途径，（ii）可以导致生物标志物的开发，对SCLC的不同亚组进行分类，以及（iii）可以提供有关联合治疗方法的信息，可以进一步研究这些方法在SCLC中的有效性。蛋白质组学分析提供了SCLC生物学的额外观点，超越了基因测序或基因表达谱，因此是对SCLC生物学研究的其他重要工作的补充。为了实现项目目标，我们提出了四个目标。目的1将发展利用质谱技术定量表征激酶组和其他ATP结合蛋白的方法。目的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。目的3将使用蛋白质印迹和反相蛋白质阵列验证ABPP的发现。目的4探讨利用RNAi和小分子抑制剂对ABPP鉴定的蛋白激酶的功能意义，以更好地指导生物标志物的选择。

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.