Selectively Targeting Oncogenic NRAS in Cancer

选择性靶向癌症中的致癌 NRAS

基本信息

批准号：
9040123
负责人：
KEVIN M. SHANNON
金额：
$ 55.05万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-04-01 至 2020-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9040123
关键词：
Address Alleles Amino Acids Biochemical Biochemistry Biology Cancer Burden Cancer cell line Cells Characteristics Chemicals Collaborations Cysteine Data Development Dominant-Negative Mutation Enzymatic Biochemistry Enzymes Frequencies Generations Genes Genetic Goals Growth HRAS gene Health Hematologic Neoplasms Hematopoietic Human Knock-in Knock-in Mouse Malignant - descriptor Malignant Neoplasms Methods Molecular Molecular Biology Techniques Mouse Strains Mus Mutant Strains Mice Mutate Mutation NRAS gene Oncogenes Oncogenic Output Pathogenesis Pathway interactions Play Post-Translational Protein Processing Protein Isoforms Proteins Proto-Oncogenes Reagent Reporting Research Personnel Serine Serine Hydrolase Signal Transduction Staging Synthesis Chemistry TAP2 gene Tertiary Protein Structure Testing Therapeutic activity-based protein profiling base cancer cell cancer type cell growth conditional mutant design drug discovery gain of function improved in vivo inhibitor/antagonist leukemia mutant novel novel strategies novel therapeutic intervention palmitoylation pre-clinical progenitor protein function ras GTPase-Activating Proteins ras Proteins research study small molecule libraries therapeutic target trafficking

项目摘要

DESCRIPTION (provided by applicant): RAS proto-oncogenes are mutated in ~30% of human cancers, but no mechanism-based treatments exist for reversing the biochemical output of oncogenic Ras proteins, which are exceedingly difficult targets for rational drug discovery. Our long-term goal is to implement mechanistic strategies to selectively inhibit the growth of cancers with somatic RAS mutations. In this project, we will investigate the Ras palmitoylation/depalmitoylation cycle, which regulates the subcellular trafficking of the N-Ras, H-Ras, and K-Ras4a isoforms, as a therapeutic target for selectively inhibiting the growth of malignancies with oncogenic NRAS mutations. Acyl protein thioesterase 1 and 2 (APT1 and APT2) catalyze N-Ras depalmitoylation and we found that "first generation" chemical inhibitors designed to inhibit these enzymes (Palmostatin B and Palmostatin M) selectively reduced the growth of primary hematopoietic progenitors and leukemia cells expressing oncogenic N-RasG12D. However, our recent studies also infer the existence of additional biochemical targets of these compounds that are essential for the growth of NRAS mutant cancer cells. This project involves a cross-disciplinary collaboration that brings together investigators with extensive expertise in synthetic chemistry (Dr. Howell), enzymology and chemical biology (Dr. Cravatt), and hematologic cancer, Ras signaling, and preclinical therapeutics (Dr. Shannon). We have collaborated to generate extensive preliminary data and novel reagents, which we will use to pursue the goals of: (1) identifying additional biochemical targets of the palmostatins; (2) developing new chemical inhibitors with improved potency and selectivity for palmostatin targets; (3) using these inhibitors combined with genetic methods to discern the relevant enzyme(s) that regulate N-Ras depalmitoylation in cancer cells; and, (4) utilizing human cancer cell lines and a new strain of mice to interrogate the palmitoylation/depalmitoylation cycle as a therapeutic target in early stage and advanced NRAS-mutant cancers. We will address these questions through two highly integrated specific aims. In Aim 1, we will identify additional biochemical targets of palmostatin M, design and characterize new chemical inhibitors, and evaluate the efficacy of these compounds in cancer cells with NRAS mutations. In Aim 2, we will utilize a novel strain of NrasG12D,C181S "knock in" mice to ask if the palmitoylation/depalmitoylation cycle is required for the growth of oncogenic Nras-driven cancers in vivo. These studies will rigorously assess the importance of the palmitoylation/depalmitoylation cycle and inform the development of new therapeutic strategies for cancers with oncogenic RAS mutations.

描述（由申请人提供）：RAS 原癌基因在约 30% 的人类癌症中发生突变，但不存在基于机制的治疗方法来逆转致癌 Ras 蛋白的生化输出，这是我们长期以来合理药物发现的极其困难的目标。长期目标是实施机制策略来选择性抑制具有体细胞 RAS 突变的癌症的生长。在这个项目中，我们将研究 Ras 棕榈酰化/去棕榈酰化。循环，调节 N-Ras、H-Ras 和 K-Ras4a 亚型的亚细胞运输，作为选择性抑制具有致癌 NRAS 突变的恶性肿瘤生长的治疗靶点（APT1 和 APT2）。催化 N-Ras 去棕榈酰化，我们发现“第一代”化学抑制剂旨在抑制这些酶（Palmostatin B 和Palmostatin M）选择性地减少了表达致癌性 N-RasG12D 的初级造血祖细胞和白血病细胞的生长。然而，我们最近的研究还推断这些化合物存在对 NRAS 突变癌细胞生长至关重要的其他生化靶标。涉及跨学科合作，汇集了在合成化学（Howell 博士）、酶学和化学生物学（Cravatt 博士）以及血液癌症方面拥有广泛专业知识的研究人员， Ras 信号传导和临床前治疗（Shannon 博士）。我们合作生成了大量的初步数据和新颖的试剂，我们将利用这些数据来实现以下目标：(1) 确定棕榈抑素的其他生化靶点；(2) 开发新的棕榈抑素靶点。对棕榈抑素靶点具有改进效力和选择性的化学抑制剂；(3) 使用这些抑制剂与遗传方法相结合来识别调节癌细胞中 N-Ras 去棕榈酰化的相关酶；利用人类癌细胞系和一种新的小鼠品系来研究棕榈酰化/去棕榈酰化循环作为早期和晚期 NRAS 突变癌症的治疗靶点，我们将通过两个高度整合的具体目标来解决这些问题。确定palmostatin M的其他生化靶点，设计和表征新的化学抑制剂，并评估这些化合物在具有NRAS突变的癌细胞中的功效。在目标2中，我们将利用一种新的菌株。 NrasG12D、C181S“敲入”小鼠，询问体内 Nras 驱动的致癌癌症的生长是否需要棕榈酰化/去棕榈酰化循环。这些研究将严格评估棕榈酰化/去棕榈酰化循环的重要性，并为新疗法的开发提供信息。针对具有致癌 RAS 突变的癌症的策略。