ACTIVITY-BASED KINASE DISCOVERY IN SMALL CELL LUNG CANCER

描述(申请人提供)：小细胞肺癌(SCLC)是一种研究不足的癌症，还没有显示出分子靶向治疗方法的使用。在这里，我们建议使用一个化学生物学平台，基于活性的蛋白质图谱(ABPP)，来研究小细胞肺癌(SCLC)的蛋白质组。ABPP使用针对酶的活性部位的化学探针来询问生物样品中酶的功能状态。利用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来根据全球蛋白激酶图谱更好地对小细胞肺癌进行分类和分组，这可以指导未来开发靶向治疗的尝试。本项目的假设是，基于Kinome的ABPP可以检测小细胞肺癌中的功能激酶，并通过以下方式指导未来的临床治疗：(1)识别作为小细胞肺癌生长和生存潜在驱动因素的活性激酶和通路；(2)可以导致对小细胞肺癌不同亚群进行分类的生物标记物的开发；(3)可以为进一步研究小细胞肺癌的有效性而提供联合治疗方法的信息。蛋白质组学提供了对小细胞肺癌生物学的额外观察，超越了基因测序或基因表达谱，因此是对研究小细胞肺癌生物学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。目标1将开发定量方法，使用质谱学来表征Kinome和其他ATP结合蛋白。目的2将使用ABPP来表征小细胞肺癌肿瘤和细胞系中的激酶和其他ATP结合蛋白。AIM 3将使用Western blotting和反相蛋白质阵列来验证ABPP发现的结果。目的4将使用RNAi和小分子抑制剂来询问ABPP识别的蛋白激酶的功能意义，以更好地指导生物标记物的选择。

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.