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（由适用提供）：该研究项目旨在全面确定小鼠和人类胰腺癌中致癌性KRAS下游的信号通路依赖性。迄今为止，KRAS本身已被证明是直接药物靶向的难治性，但一系列潜在的可药物信号传导效应传达了突变体KRAS下游的信号传导。这些效果包括：1）RAF/MEK/ERK激酶级联反应； 2）由PLC/PKC和PI3K/AKT介导的磷脂信号传导； 3）rala/b和rac1介导的GTPase信号传导。这些下游效应的一小部分已在胰腺癌前体的主动性中进行了审问，以表明必要的作用。但是，在维持既定的胰腺癌中的必要作用方面，都没有对没有一定的评估，从而在我们合理规划这些途径的靶向治疗的能力方面留下了很大的差距。因此，我们建议在小鼠和人类模型系统中遗传剖析这些可能吸毒的下游介体。该项目的中心假设是：首先，突变体KRAS引起的复合表型的个体成分将取决于不同的下游信号传导介体；其次，下游介质的遗传和药物靶向组合将揭示新的治疗脆弱性。第三，不同突变的KRAS等位基因可能对单个信号通路的治疗破坏敏感。为了检验这些假设，我们提出了以下特定目的：1）创建新的GEMM ESC基于鼠模型，允许在已建立的胰腺肿瘤中raf1，pik3ca，plcε，rala/b和rac1的定时失活； 2）使用人类胰腺癌3D器官培养系统结合不同下游KRAS介体的遗传和药物抑制，以鉴定新的治疗协同和脆弱性； 3）创建和比较涉及KRAS G12R和Q61H的新的基于ESC的基于ESC的基因工程鼠标。该项目利用高度创新的“快速” Gemm ESC小鼠技术，包括新的专用胚泡注射设施，以及用于患者衍生的肿瘤器官3D器官培养的新技术。总之，这些研究将对既定的KRAS驱动胰腺癌中下游信号依赖性进行全面分析。我们预计这些研究将为未来的临床试验设计，其中利用这些依赖性以获得治疗。