Systematic identification of oncogenic KRAS synthetic lethal interactions

系统鉴定致癌 KRAS 合成致死相互作用

• 批准号: 9150537
• 负责人: William C. Hahn
• 金额: $ 80.78万
• 依托单位: BROAD INSTITUTE, INC.
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-25 至 2019-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9150537
• 关键词: 1-Phosphatidylinositol 3-Kinase; Adverse effects; Alleles; Animal Model; Animals; Binding; Biological Assay; Biological Models; Biological Process; CRISPR library; CRISPR screen; CRISPR/Cas technology; Cancer Model; Cancer cell line; Cell Death; Cell Line; Cells; Cessation of life; Clinic; Clinical Trials; Clustered Regularly Interspaced Short Palindromic Repeats; Colon Carcinoma; Dependency; Development; Diagnosis; Dose-Limiting; Essential Genes; Exhibits; Experimental Models; Foundations; Gene Expression; Generations; Genes; Genetic; Genetic Engineering; Goals; Health; Human; In Vitro; KRAS2 gene; MAP Kinase Gene; MEK inhibition; Maintenance; Malignant Neoplasms; Malignant neoplasm of lung; Malignant neoplasm of pancreas; Mediating; Methods; Mitogen-Activated Protein Kinase Inhibitor; Mus; Mutate; Mutation; Oncogenes; Oncogenic; Organoids; Pathogenesis; Pathway interactions; Patients; Phenotype; Phosphotransferases; Process; Protein-Serine-Threonine Kinases; Proteins; RNA Interference; Signal Pathway; Signal Transduction; System; TBK1 gene; Technology; Therapeutic Agents; Toxic effect; Tumorigenicity; Work; Xenograft procedure; base; curative treatments; effective therapy; gene product; genetic analysis; genome editing; genome-wide; in vivo; in vivo Model; inhibitor/antagonist; innovation; insight; loss of function; mouse model; mutant; new technology; novel; novel therapeutic intervention; novel therapeutics; pre-clinical; prevent; ral Guanine Nucleotide Exchange Factor; research study; resistance mechanism; translational study; tumor; tumor growth; tumor microenvironment; tumor progression

 DESCRIPTION (provided by applicant): Activating mutations of KRAS are among the most common mutations found in human cancers, and cancers that harbor KRAS clearly depend on the activity of this oncogene for tumor maintenance. However, despite considerable effort, direct targeting of KRAS or known KRAS effector pathways has not yet led to effective therapies in cancers that harbor mutant KRAS. An alternative approach to direct targeting of known cancer alleles is to exploit the genetic concept of synthetic lethality, in which gene products are identified that, when suppressed or inhibited, result in cell death only in the presence of another non-lethal mutation. Synthetic phenotype screens in model organisms have provided insights into a broad spectrum of biological processes and in principle; this strategy provides a means to target currently "undruggable" proteins while simultaneously reducing the potential for side effects. Over the past several years, we and others have used RNAi-mediated suppression of gene expression to identify genes whose expression is required in cell lines that depend on mutant KRAS for survival. Inhibitors to some of these synthetic lethal candidates are now the subject of clinical trials in KRAS-driven cancers. However, these early studies used different cells and experimental systems and were limited by scale, technological issues or context. In addition, the discovery and development of new gene manipulation technologies such as Cas9-CRISPR, and methods to isolate and propagate human tumors now provide the opportunity to comprehensively identify novel genes and pathways that are required for the survival of KRAS-dependent cancers. In this application, we propose to use new genome scale gene manipulation technologies, potentially more relevant human and murine experimental models and advanced analytical approaches in an integrated approach to systematically identify KRAS synthetic lethal relationships in cell, organoid and animal models. Specifically, we will performed genome scale CRISPR mediated loss of function experiments to identify KRAS co-dependencies in both in vitro and in vivo model systems and identify genes and pathways that when inhibited synergize with known KRAS effector pathways to induce tumor regression in KRAS driven cancers. These studies will permit us to define the signaling network perturbed by oncogenic KRAS necessary for tumor maintenance and progression. Targets identified by these approaches will form the basis of translational studies to develop novel therapeutic approaches.

 描述（由适用提供）：KRAS的激活突变是人类癌症中最常见的突变之一，携带KRAS的癌症显然取决于这种癌基因对肿瘤维持的活性。然而，尽管有很多努力，但直接靶向KRAS或已知的KRAS效应途径尚未导致具有突变体Kras的癌症的有效疗法。直接靶向已知癌症等位基因的另一种方法是利用合成致死性的遗传概念，其中鉴定出基因产物在抑制或抑制时，只有在另一种存在下才会导致细胞死亡 非致命突变。模型生物中的合成表型筛选为广泛的生物过程和原则上提供了见解。该策略为靶向当前“不良”蛋白的目标提供了一种手段，同时降低了副作用的潜力。在过去的几年中，我们和其他人使用RNAi介导的基因表达抑制来鉴定其在依赖突变体KRAS生存的细胞系中所必需的基因。这些合成致死候选者中的某些抑制剂现在是KRAS驱动癌症中临床试验的主题。但是，这些早期研究使用了不同的细胞和实验系统，并受到规模，技术问题或环境的限制。此外，现在的新基因操纵技术（例如cas9-crispr）的发现和开发以及分离和传播人肿瘤的方法现在提供了机会，可以全面识别依赖Kras依赖性癌症的新型基因和途径。在此应用中，我们建议使用新的基因组量表基因操纵技术，可能更相关的人类和鼠实验模型以及综合方法中的高级分析方法，以系统地识别细胞，器官和动物模型中的KRAS合成致死关系。具体来说，我们将执行 基因组量表CRISPR介导的功能实验丢失，以鉴定体外和体内模型系统中的KRAS共依赖性，并鉴定基因和途径，当抑制已知KRAS效应途径抑制KRAS驱动器中肿瘤回归时，这些基因和途径。这些研究将使我们能够定义受肿瘤维持和进展所必需的致癌KRAS扰动的信号网络。这些方法确定的目标将构成转化研究的基础，以开发新的治疗方法。