Genetic dissection of oncogenic Kras signaling

致癌 Kras 信号传导的基因剖析

基本信息

批准号：
10441550
负责人：
Monte Meier Winslow
金额：
$ 43.79万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-07-01 至 2026-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10441550
关键词：
Alleles Area Bar Codes Biochemical Biology CRISPR/Cas technology Cancer Model Cancer Patient Cell Culture Techniques Cell Line Cessation of life Chemicals Clinical Clinical Trials Codon Nucleotides Consumption Correlative Study Data Dissection Environment Epithelial Cells Generations Genetic Genetically Engineered Mouse Genome engineering Genomics Genotype Goals Growth Histologic Human Impairment KRAS oncogenesis KRAS2 gene Lung Lung Neoplasms MEKs Malignant Neoplasms Malignant neoplasm of lung Maps Methods Modeling Molecular Mus Mutate Mutation Oncogenes Oncogenic PTPN11 gene Pathway interactions Patients Pharmacogenomics Pharmacology Physiological Positioning Attribute Proteins Research Research Personnel Signal Pathway Signal Transduction Somatic Cell System Therapeutic Therapeutic Uses Time Variant base cancer care cancer genetics cancer genome cancer therapy cancer type carcinogenesis cost drug sensitivity experimental study genetic approach genome sequencing in vivo loss of function mathematical model mouse model mutant novel overexpression repaired tool tumor tumor growth tumorigenic

项目摘要

ABSTRACT Lung cancer is a prevalent cancer type that leads to more deaths than the next four major cancer types combined. KRAS is one of the most frequent oncogenes in human lung cancer. Despite more than 30 years of biochemical and cell culture studies, as well as correlative studies on human tumors and clinical trials, therapeutic options for patients with oncogenic KRAS-driven tumors are just beginning to emerge. KRAS is often mutated at codons 12 and 13, but these mutations are diverse and these different mutant forms of KRAS have dramatically different biochemical features. By integrating conventional genetically-engineered mouse models and CRISPR/Cas9-based somatic genome engineering with quantitative genomics and mathematical modeling, we recently established CRISPR/Cas9-based approaches that enable the generation and quantitative analysis of multiple tumor genotypes in parallel in vivo. By employing homology directed repair in somatic cells, we induce a panel of oncogenic Kras variants, and uncovered an unexpectedly dramatic difference in oncogenic potential of different Kras variants in vivo. In addition to the diversity of different oncogenic KRAS variants, the compendium of important pathways downstream of oncogenic KRAS remains relatively poorly understood. The goals of this proposal are 1) to use genomic and pharmacological methods to generate a quantitative understanding of different signaling requirements in cancers driven by different Kras variants and 2) to uncover novel functional regulators of Kras-driven carcinogenesis. To understand the basis for the differential oncogenic potential of different oncogenic Kras variants, we will use our multiplexed genetic approaches to quantify the impact of increasing either overall Kras signaling or discrete downstream pathways on the in vivo tumorigenic potential of diverse oncogenic Kras variants. We will also use therapeutic treatments to uncover the requirement for sustained PI3K and Raf/Mek/Erk signaling in established lung tumors driven by diverse Kras variants. Finally, to expand our understanding of Kras-driven tumorigenesis beyond the canonical effect pathways, we will directly analyze the function of novel Kras-interacting proteins on lung tumor growth in vivo. By performing multiplexed genomic and pharmacologic analyses of oncogenic Kras signaling in cancer, we will uncover the molecular mechanisms that contribute to tumor growth driven by different variants of KRAS. We will define specific therapeutic sensitivities of lung tumors driven by diverse oncogenic mutations. Our proposed research is significant because it will uncover interesting new areas of biology, motivate genotype-directed clinical trials, and facilitate precision cancer therapy for lung cancer patients with KRAS- mutant tumors.

抽象的肺癌是一种普遍的癌症类型，导致死亡比接下来的四种主要癌症类型更多合并。 KRAS是人类肺癌中最常见的肿瘤基因之一。尽管超过30年生化和细胞培养研究，以及有关人类肿瘤和临床试验的相关研究致癌性KRAS驱动肿瘤患者的治疗选择刚刚开始出现。克拉斯是通常在密码子12和13中突变，但是这些突变是多种多样的，这些不同的突变形式的Kras 具有截然不同的生化特征。通过整合常规遗传工程的小鼠具有定量基因组学和数学的模型和基于CRISPR/CAS9的体细胞基因组工程建模，我们最近建立了基于CRISPR/CAS9的方法，以使一代和在体内并联多种肿瘤基因型的定量分析。通过采用同源性定向维修躯体细胞，我们诱导了一系列致癌的Kras变体，并发现了一个出乎意料的戏剧性体内不同KRAS变体的致癌潜力的差异。除了不同的多样性致癌性KRAS变体，致癌Kras下游重要途径的汇编仍然存在相对较少的理解。该提案的目标是1）使用基因组和药理方法在不同KRAS驱动的癌症中对不同信号要求的定量了解变体和2）揭示了KRAS驱动的致癌作用的新型功能调节剂。了解基础对于不同致癌KRAS变体的差异致癌潜力，我们将使用多重遗传量化增加总体KRAS信号传导或离散下游途径的影响的方法关于多种致癌性KRAS变体的体内致瘤潜力。我们还将使用治疗疗法揭示对已建立的肺部肿瘤中持续的PI3K和RAF/MEK/ERK信号传导的要求多样化的Kras变体。最后，将我们对KRAS驱动肿瘤发生的理解扩展到规范之外效应途径，我们将直接分析新型KRAS相互作用蛋白对肺部肿瘤生长的功能体内。通过对癌症中的致癌KRAS信号传导进行多路复用基因组和药理分析，我们将发现由不同变体驱动的肿瘤生长的分子机制克拉斯。我们将定义由多种致癌突变驱动的肺肿瘤的特定治疗敏感性。我们提出的研究很重要，因为它将发现有趣的新生物学领域，激励基因型定向的临床试验，并促进肺癌患者的精确癌症治疗突变肿瘤。