Targeting the kinome in K-ras driven colorectal cancers

项目摘要/摘要 RAS蛋白的突变，特别是K-ras，在人类癌症中是常见的，信号通路 已经对这种癌蛋白进行了详细的检测。尽管做出了密集的努力来翻译这篇文章 尽管K-ras基因突变肿瘤对癌症治疗的认识有所改善，但它在科学和临床上仍是一个艰巨的挑战。 在这项建议中，我们重点研究在结直肠癌(CRC)中常见的个别KRAS突变， 因为这些突变与对靶向抑制剂的不同敏感性有关，因此可能与 不同的信号程序，需要个性化的治疗策略。访问和利用这些截然不同的 计划，我们采用了一种创新的基于质谱学的技术，使大多数人类 同时分析Kinome，提供了在无偏倚的情况下探索K-ras诱导的Kinome的手段 时尚。我们独特的方法结合了质谱学和蛋白激酶捕获珠子来监测 从肿瘤细胞激活的蛋白激酶，提供了肿瘤激活酶活性的“全景”。使用这个 技术，在目标1中，我们将在(I)模型等基因CRC系统中测量Kinome的基本活性，该系统 代表常见的与CRC相关的KRAS突变等位基因和(Ii)一组特征良好的人类KRAS 突变的CRC细胞系，KRAS基因敲除前后，以确定其活性依赖于 该癌基因的各种突变形式的表达。在目标2中，我们将探讨儿童权利公约的复原力。 KRAS kinome(S)对MEK的抑制，以识别和靶向促进耐药性的补偿激酶。在AIM 3、我们将利用这些信息在体内进行有针对性的合成致死筛选，测试其有效性 在依赖KRAS的结直肠癌移植瘤和结直肠癌敲入小鼠中抑制这些激活的蛋白激酶 由常见的KRAS外显子2(G12D)或外显子4(A146T)突变驱动的模型。通过这种方式，我们希望 确定在治疗这些常见类型的K-ras时有效的个体化激酶抑制剂组合- 驱动CRC。

PROJECT SUMMARY/ABSTRACT Mutations in RAS proteins, in particular K-ras, are common in human cancer, and the signaling pathways emanating from this oncoprotein have been examined in detail. Despite intensive efforts to translate this knowledge into improved cancer care, K-ras mutant tumors remain a formidable scientific and clinical challenge. In this proposal, we focus on individual KRAS mutations that are commonly found in colorectal cancer (CRC), because these mutations are associated with different sensitivities to targeted inhibitors and thus may engage distinct signaling programs and require individualized therapeutic strategies. To access and exploit these distinct programs, we employ an innovative mass spectrometry-based technique that enables the majority of the human kinome to be analyzed simultaneously, providing the means to explore the K-ras induced kinome in an unbiased fashion. Our unique approach combines mass spectrometry and protein kinase-capture beads to monitor activated protein kinases from tumor cells, providing the ‘big picture’ of tumor kinase activity. Using this technology, in Aim 1 we will measure the basal activity of the kinome in (i) a model isogenic CRC system that represents common CRC associated KRAS mutant alleles and (ii) a panel of well characterized human KRAS mutant CRC cell lines, before and after KRAS knockdown, to identify kinases whose activity is dependent on expression of the various mutant forms of this oncogene. In Aim 2, we will explore the resiliency of the CRC KRAS kinome(s) to MEK inhibition to identify and target compensatory kinases that promote resistance. In Aim 3, we will use this information to carry out a targeted synthetic lethal screen in vivo, testing the effectiveness of inhibiting these activated protein kinases in KRAS dependent CRC xenografts and in CRC knock-in mouse models driven by a common KRAS exon 2 (G12D) or an exon 4 (A146T) mutation. In this way, we hope to identify individualized kinase inhibitor combinations that are effective in treating these common types of K-ras- driven CRC.