Comprehensive genetic dissection of druggable KRAS targets

可药物 KRAS 靶点的全面基因剖析

基本信息

批准号：
9476973
负责人：
Steven D Leach
金额：
$ 36.99万
依托单位：
DARTMOUTH COLLEGE
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-05-01 至 2021-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9476973
关键词：
3-Dimensional Alleles Attention Biological Models Biology Cancer Biology Cancer Etiology Cessation of life Clinical Clinical Trials Colorectal Cancer Data Dependence Disease Dissection Drug Industry Evolution Future Genetic Genetically Engineered Mouse Goals Growth Guanosine Triphosphate Phosphohydrolases Human Individual Injections KRAS2 gene MEKs Maintenance Malignant neoplasm of pancreas Malignant neoplasm of prostate Mediating Mediator of activation protein Memorial Sloan-Kettering Cancer Center Molecular Mus Mutation Oncogenic Organoids Outcome PI3K/AKT Pathway interactions Patients Pharmacology Phenotype Phospholipids Phosphotransferases Production RALA gene Refractory Research Project Grants Role Signal Pathway Signal Transduction System Techniques Technology Testing Therapeutic Time Treatment Efficacy Tumor-Derived United States base blastocyst cohort design effective therapy embryonic stem cell human model in vitro Model inhibitor/antagonist innovation knock-down malignant breast neoplasm mouse model mutant novel novel therapeutics pancreatic neoplasm public health relevance synergism therapeutic target tumor

项目摘要

DESCRIPTION (provided by applicant): This research project seeks to comprehensively determine signaling pathway dependencies downstream from oncogenic KRAS in mouse and human pancreatic cancer. While KRAS itself has so far proven refractory to direct pharmacologic targeting, an array of potentially druggable signaling effectors convey signaling downstream from mutant KRAS. These effectors include: 1) the RAF/MEK/ERK kinase cascade; 2) Phospholipid signaling mediated by PLC/PKC and PI3K/AKT; and 3) GTPase signaling mediating by RALA/B and RAC1. A small subset of these downstream effectors have been interrogated for requisite roles in the initiation of pancreatic cancer precursors. However, none have been genetically evaluated with respect to a requisite role in the maintenance of established pancreatic cancer, leaving substantial gaps in our ability to rationally plan therapeutic targeting of these pathways. We therefore propose to genetically dissect these potentially druggable downstream mediators in both mouse and human model systems. The central hypotheses of this project are: First, that the individual components of the composite phenotype induced by mutant Kras will be dependent upon different downstream signaling mediators; second, that combined genetic and pharmacologic targeting of downstream mediators will reveal novel therapeutic vulnerabilities; and third, that different mutant Kras alleles may be differentially sensitive to therapeutic disruption of individual signaling pathways. To test these hypothesis, we propose the following Specific Aims: 1) To create new GEMM ESC-based murine models allowing for the timed inactivation of Raf1, Pik3ca, PLCε, RalA/B and Rac1 in established pancreatic tumors; 2) To combine genetic and pharmacologic inhibition of different downstream KRAS mediators to identify new therapeutic synergies and vulnerabilities using a human pancreatic cancer 3D organoid culture system; and 3) To create and compare new ESC-based genetically engineered mouse models involving Kras G12R and Q61H. The project leverages highly innovative "Speedy" GEMM ESC mouse technology, including a new dedicated blastocyst injection facility, as well as novel techniques for 3D organoid culture of patient-derived tumor organoids. Together, these studies will for the first tim provide a comprehensive analysis of downstream signaling dependencies in established Kras-driven pancreatic cancer. We anticipate that these studies will inform the design of future clinica trials in which these dependencies are exploited for therapeutic gain.

描述（由申请人提供）：该研究项目旨在全面确定小鼠和人类胰腺癌中致癌 KRAS 下游信号通路的依赖性，尽管迄今为止已证明 KRAS 本身难以直接药理学靶向，但一系列潜在的可药物化信号传导效应器可传递信号传导。这些效应器包括：1) RAF/MEK/ERK 激酶级联；2) 由 PLC/PKC 介导的磷脂信号传导。 PI3K/AKT；以及 3) 由 RALA/B 和 RAC1 介导的 GTPase 信号转导已被研究，以了解其在胰腺癌前体发生过程中的必要作用。在已建立的胰腺癌的维持中发挥着必要的作用，因此我们在合理规划这些途径的治疗靶向的能力方面存在很大差距，因此我们建议从基因角度剖析这些潜在的可药物化的下游介质。在小鼠和人类模型系统中，该项目的中心假设是：首先，突变体 Kras 诱导的复合表型的各个成分将依赖于不同的下游信号传导介质；其次，结合下游介质的遗传和药理学靶向。将揭示新的治疗漏洞；第三，不同的突变体 Kras 等位基因可能对个别信号通路的治疗破坏敏感。为了检验这些假设，我们提出以下具体目标：1）创建新的基于 GEMM ESC 的小鼠模型，允许在已建立的胰腺肿瘤中定时灭活 Raf1、Pik3ca、PLCε、RalA/B 和 Rac1；使用人类胰腺癌 3D 类器官培养系统对不同下游 KRAS 介质进行药理抑制，以确定新的治疗协同作用和脆弱性； 3) 创建并比较涉及 Kras G12R 和 Q61H 的新型 ESC 基因工程小鼠模型该项目利用高度创新的“Speedy”GEMM ESC 小鼠技术，包括新的专用囊胚注射设施以及 3D 类器官培养新技术。这些研究将首次对已确定的 Kras 驱动的胰腺癌下游信号依赖性进行全面分析。为未来临床试验的设计提供信息，其中利用这些依赖性来获得治疗效果。