Systematic shRNA screens for the analysis of critical signaling pathways in KRAS-mutant multiple myeloma

系统性 shRNA 筛选分析 KRAS 突变型多发性骨髓瘤的关键信号通路

• 批准号: 242271643
• 负责人: Professor Dr. Martin Eilers
• 金额: --
• 依托单位: Lehrstuhl für Biochemie und Molekularbiologie
• 依托单位国家: 德国
• 项目类别: Clinical Research Units
• 财政年份: 2013
• 资助国家: 德国
• 起止时间: 2012-12-31 至 2015-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/242271643?language=en
• 关键词: Systematic; shRNA; screens; analysis; critical

Mutations in KRAS are driver mutations in multiple myeloma (MM) yet no small moleculesthat target the encoded protein (KRAS) are available. Mutant KRAS activates multiple signal transduction pathways that act in unknown and potentially highly cell-specific combinations to drive growth of MM cells. In the previous funding period, we have established systematic loss-of-function (shRNA) screens as lentiviral pool screens coupled with next generation sequencing to a complexity of 10,000 shRNAs. We found that these screens can identify therapeutically relevant co-operations between downstream effector pathways of mutant KRAS. The applied-for project aims to firmly establish and validate this concept using different MM cell lines. Comparison with genome sequence data will establish whether specific biomarkers (for example mutations) reliably indicate co-operativity between critical signaling pathways. In parallel, we propose to design and clone a shRNA library that targets critical effector pathways with limited complexity in order to allow the analysis of KRASdependent signaling pathways in small cell populations (for example, in in vivo models of tumorigenesis).

KRAS突变是多发性骨髓瘤(MM)的驱动突变，但没有针对编码蛋白(KRAS)的小分子可用。突变的KRAS激活多个信号转导通路，这些信号转导通路在未知的和潜在的高度细胞特异性的组合中发挥作用，从而推动MM细胞的生长。在之前的资助期间，我们已经建立了系统性功能丧失(ShRNA)筛查，作为慢病毒池筛查，加上下一代测序，复杂性为10,000个shRNA。我们发现，这些筛选可以识别突变KRAS下游效应通路之间的治疗相关合作。申请的项目旨在利用不同的MM细胞系坚定地建立和验证这一概念。与基因组序列数据的比较将确定特定的生物标记物(例如突变)是否可靠地表明关键信号通路之间的协同作用。同时，我们建议设计和克隆一个针对关键效应通路的shRNA文库，其复杂性有限，以便能够分析小细胞群体中依赖KRASs的信号通路(例如，体内肿瘤发生的模型)。