Interrogating the RAS interactome for therapeutic vulnerabilities

询问 RAS 相互作用组的治疗漏洞

基本信息

批准号：
10732791
负责人：
Hema Adhikari
金额：
$ 24.82万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-03-10 至 2025-12-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10732791
关键词：
Ablation Antineoplastic Agents Automobile Driving Binding Biochemical Biological Assay CRISPR/Cas technology Cell Fractionation Clinical Cultured Cells Data Data Set Dependence Disease Drug Targeting Enhancers Future Genes Genetic Genetically Engineered Mouse Genotype Goals Grant Homeostasis Human Immunotherapy K-ras mouse model KRAS oncogenesis KRAS2 gene Label Libraries Lipids MAP Kinase Gene Malignant Neoplasms Mediating Mediator Membrane Mining Mutate Mutation Nature Oncogenes Oncogenic Oncoproteins Oxidation-Reduction Oxidative Stress Oxidative Stress Induction Pancreas Pathway interactions Pentosephosphate Pathway Pharmaceutical Preparations Phase Phosphatidylinositol Phosphates Phosphatidylinositols Phosphotransferases Prevalence Process Protein Isoforms Proteins RAS driven cancer RAS genes RAS inhibition Ras Inhibitor Research Research Personnel Role Signal Transduction Signaling Protein Stress Systems Biology Techniques Testing Therapeutic Training Validation Xenograft procedure cancer cell cell growth cell transformation experimental study inhibitor inorganic phosphate mouse model mutant new therapeutic target novel novel therapeutics pancreatic cancer model pharmacologic phosphoproteomics ras Oncogene ras Proteins response small molecule inhibitor therapeutic target tumor growth tumor xenograft tumorigenesis tumorigenic

项目摘要

PROJECT SUMMARY AND ABSTRACT The RAS genes KRAS, NRAS, or HRAS, are commonly mutated in human cancers. Clinically inhibiting RAS has proven challenging and RAS-mutant cancers remain some of the most intractable diseases, even to immunotherapies. It is thus critical to elucidate oncogenic RAS signaling, not only to better understand the tumorigenic process, but also to identify new potential therapeutic targets. To this end, I exploited the novel technique of BirA-mediated proximity labeling to identify proteins within the immediate vicinity (interactome) of each RAS isoform. I then screened an sgRNA library targeting interactome components for genes promoting RAS transformed cell growth, identifying the druggable phosphatidylinositol phosphate lipid kinase PIP5K1A as specifically driving KRAS oncogenesis. PIP5K1A represents an entirely new therapeutic target in KRAS-mutant cancers, and suggests that other proteins in the RAS interactome may similarly mediate RAS oncogenesis. I will capitalized on these discoveries in three aims. As PIP5K1A is a druggable kinase it provides a way to specifically inhibit KRAS oncogenesis, which could be exploited to enhance the antineoplastic activity of drugs targeting RAS effector pathways. Thus, in aim 1 I will elucidate the role and therapeutic potential of targeting PIP5K1A in KRAS-mutant cancers. The identification of PIP5K1A promoting KRAS oncogenesis suggests that other interactome proteins may similarly mediate RAS function. Thus, in aim 2 I will mine the RAS interactome for novel modifiers of RAS oncogenesis, focusing on the interactome protein EFR3A as a potential general mediator of oncogenic RAS-driven tumorigenesis. Finally, the RAS interactome is most certainly dynamic, varying under different conditions. Determining the content of the RAS interactome under distinct settings may thus identify new vulnerabilities specific to diverse cellular conditions. Thus, in aim 3 I will probe the RAS interactome in response to cellular perturbations. In sum, I will expand upon my discovery that PIP5K1A promotes KRAS oncogenesis to explore this kinase as a new therapeutic target and identify other novel therapeutic vulnerabilities that exists within the RAS interactome. The K99 segment of this grant will complete my training in RAS signal transduction, extend my training into phosphoproteomics, xenograft and genetically engineered mouse models of tumorigenesis. The R00 segment will capitalize on the use of proximity labeling to study the dynamic nature of oncogenic RAS signaling. My long-term goal is to transition into an independent investigator and apply systems biology approaches to uncover the signaling circuitry of oncogene drivers with the objective of identifying novel therapeutic vulnerabilities in RAS-mutant cancers.

项目概要和摘要 RAS 基因 KRAS、NRAS 或 HRAS 在人类癌症中通常发生突变。临床上抑制 RAS 已被证明具有挑战性，RAS 突变癌症仍然是一些最棘手的疾病，即使对于免疫疗法也是如此。因此，阐明致癌 RAS 信号传导至关重要，不仅可以更好地了解致瘤过程，而且可以确定新的潜在治疗靶点。为此，我利用 BirA 介导的邻近标记新技术来识别每个 RAS 异构体紧邻区域（相互作用组）内的蛋白质。然后，我筛选了一个针对相互作用组成分的 sgRNA 文库，以寻找促进 RAS 转化细胞生长的基因，确定可药物化的磷脂酰肌醇磷酸脂激酶 PIP5K1A 专门驱动 KRAS 肿瘤发生。 PIP5K1A 代表了 KRAS 突变癌症的全新治疗靶点，并表明 RAS 相互作用组中的其他蛋白质可能类似地介导 RAS 肿瘤发生。我将利用这些发现实现三个目标。由于 PIP5K1A 是一种可药物激酶，它提供了一种特异性抑制 KRAS 肿瘤发生的方法，可用于增强针对 RAS 效应通路的药物的抗肿瘤活性。因此，在目标 1 中，我将阐明靶向 PIP5K1A 在 KRAS 突变癌症中的作用和治疗潜力。 PIP5K1A 促进 KRAS 肿瘤发生的鉴定表明其他相互作用蛋白可能类似地介导 RAS 功能。因此，在目标 2 中，我将挖掘 RAS 相互作用组以寻找 RAS 肿瘤发生的新修饰剂，重点关注相互作用组蛋白 EFR3A 作为致癌 RAS 驱动的肿瘤发生的潜在一般介质。最后，RAS 相互作用组肯定是动态的，在不同条件下会发生变化。因此，确定不同设置下 RAS 相互作用组的内容可能会识别特定于不同细胞条件的新漏洞。因此，在目标 3 中，我将探测 RAS 相互作用组以响应细胞扰动。总之，我将扩展 PIP5K1A 促进 KRAS 肿瘤发生的发现，探索该激酶作为新的治疗靶点，并确定 RAS 相互作用组内存在的其他新的治疗漏洞。这笔资助的 K99 部分将完成我在 RAS 信号转导方面的培训，并将我的培训扩展到磷酸蛋白质组学、异种移植和肿瘤发生的基因工程小鼠模型。 R00 部分将利用邻近标记来研究致癌 RAS 信号传导的动态性质。我的长期目标是转型为一名独立研究者，并应用系统生物学方法来揭示癌基因驱动因素的信号通路，从而确定 RAS 突变癌症的新治疗漏洞。