Investigating functional interactions with KRAS G12C inhibition

研究 KRAS G12C 抑制的功能相互作用

• 批准号: 9761082
• 负责人: Kevin Lou
• 金额: $ 3.98万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-03-01 至 2022-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9761082
• 关键词: 3-Dimensional; Alleles; Binding; Biology; CRISPR interference; Cancer cell line; Cell Line; Cell Proliferation; Cell Survival; Chemical Agents; Chemicals; Clinic; Clinical; Clinical Trials; Codon Nucleotides; Combined Modality Therapy; Complement; Cysteine; Data; Data Set; Dependence; Development; Drug Sensitization; Epidermal Growth Factor Receptor Tyrosine Kinase Inhibitor; Genes; Genetic; Genomic approach; Goals; Growth; Guanosine; Guanosine Triphosphate; Human; In Vitro; Inherited; Investigation; KRAS2 gene; Knowledge; Light; Malignant Neoplasms; Maps; Mediating; Missense Mutation; Modeling; Mutate; Mutation; NF1 gene; Neurofibromatosis Type 1 Protein; Non-Small-Cell Lung Carcinoma; Oncogenes; Oncogenic; Pathway interactions; Patients; Pharmacology; Phenotype; Play; Point Mutation; Predictive Value; Problem Solving; Proteins; Receptor Protein-Tyrosine Kinases; Regulation; Research; Resistance; Role; Signal Transduction; Syndrome; Therapeutic; Translating; Tumor Suppressor Proteins; Tyrosine Kinase Inhibitor; clinically relevant; functional genomics; gene repression; genetic resistance; genome-wide; genomic data; genomic platform; in vivo; inhibitor/antagonist; insight; knock-down; mutant; mutational status; novel; nuclease; outcome forecast; pre-clinical; programs; response; targeted agent; targeted treatment; therapeutic target; tumor

Project Summary / Abstract Mutational activation of KRAS represents the most frequently occurring oncogenic driver across human cancers and is associated with poor prognosis and resistance to treatment. Therapeutic options for patients with KRAS mutations have been limited by the lack of direct KRAS inhibitors in clinical trials – therefore efforts to date have focused on searching for therapies that exploit the principle of synthetic lethality or inhibit downstream signaling cascades activated by KRAS mutations. Genome-wide functional genomics screens have been used to identify synthetic lethal (SL) vulnerabilities associated with KRAS mutant tumors with the hope of identifying singular targets to treat KRAS mutant tumors. Although these functional genomics efforts have contributed to a rich understanding of the genetic dependencies associated with KRAS mutations, such findings have not yet translated to clinical or pre-clinical therapeutic benefit. With the recent emergence of well-characterized, potent, in vivo active direct KRASG12C inhibitors (ARS-1620), it is now possible to directly inhibit this driver-oncogene pharmacologically in an allele-specific manner. The goal of this proposal is to use the novel KRASG12C inhibitor ARS-1620 in a genome-wide assessment of genetic dependencies that complements existing approaches. Aim 1 proposes to identify genes that are responsible for mediating sensitivity and resistance to KRASG12C inhibition using a genome-wide nuclease-dead Cas9-mediated transcriptional repression (CRISPRi) functional genomics platform. Combination therapies targeting KRASG12C can then be derived from knowledge of factors that increase sensitivity to the KRASG12C inhibitor. Preliminary results from CRISPRi screens have raised additional questions regarding the dynamic intracellular regulation of KRASG12C that warrant further investigation. Aim 2 proposes to assess the interactions between receptor tyrosine kinases (RTKs) and KRASG12C in supporting an aberrantly activated GTP state. Additionally, the possibility of therapeutically targeting KRAS mutant cancers with RTK inhibitors will be assessed. Aim 3 proposes to understand the mechanism by which knockdown of the GTPase activating protein (GAP) NF1 confers resistance to KRASG12C inhibition. This result came as a surprise as the tumor suppressor role of NF1 is thought to be negated by KRAS mutation at codon 12. Completion of this proposal will deepen an understanding of mutant KRAS genetic dependencies complementary to existing approaches, inform the development of KRASG12C-targeted combination therapies, and shed light on a potentially novel function of a clinically relevant tumor suppressor.

项目总结/摘要 KRAS的突变激活是人类癌症中最常见的致癌驱动因素 并且与不良预后和对治疗的抵抗有关。KRAS患者的治疗选择 由于临床试验中缺乏直接的KRAS抑制剂，因此迄今为止的努力已经受到限制。 专注于寻找利用合成致死原理或抑制下游信号传导的疗法 KRAS突变激活的级联反应。全基因组功能基因组学筛选已用于鉴定 与KRAS突变型肿瘤相关的合成致死（SL）漏洞，希望能够识别出 治疗KRAS突变型肿瘤的靶点。尽管这些功能基因组学的努力为丰富的 了解与KRAS突变相关的遗传依赖性，这些发现还没有 转化为临床或临床前治疗益处。随着最近出现的特点鲜明，有力， 体内活性直接KRASG 12 C抑制剂（ARS-1620），现在可以直接抑制这种驱动致癌基因 以等位基因特异的方式进行。该提案的目标是使用新的KRASG 12 C抑制剂 ARS-1620在全基因组评估遗传依赖性，补充现有的方法。目的 1提出鉴定负责介导对KRASG 12 C抑制的敏感性和抗性的基因 使用全基因组核酸酶死亡Cas9介导的转录抑制（CRISPRi）功能基因组学 平台靶向KRASG 12 C的联合疗法然后可以从增加KRASG 12 C表达的因素的知识中获得。 对KRASG 12 C抑制剂敏感。CRISPRi筛选的初步结果引发了其他问题 关于KRASG 12 C的动态细胞内调节，值得进一步研究。目标2建议， 评估受体酪氨酸激酶（RTK）和KRASG 12 C之间的相互作用， 激活GTP状态。此外，使用RTK治疗靶向KRAS突变型癌症的可能性 将评估抑制剂。目的3提出了解GT3基因敲低的机制， 活化蛋白（GAP）NF 1赋予对KRASG 12 C抑制的抗性。这一结果令人惊讶，因为 NF 1的肿瘤抑制作用被认为被密码子12处的KRAS突变否定。完成本 一项提案将加深对突变KRAS遗传依赖性的理解， 方法，为KRASG 12 C靶向联合治疗的开发提供信息，并揭示了潜在的 临床相关肿瘤抑制因子的新功能。