Defining the role of Mst1/Mst2 in regulating metabolic alterations in Ras driven NSCLC

定义 Mst1/Mst2 在调节 Ras 驱动的 NSCLC 代谢改变中的作用

基本信息

批准号：
10323278
负责人：
Jennifer Bailey Lundberg
金额：
$ 17.98万
依托单位：
UNIVERSITY OF TEXAS HLTH SCI CTR HOUSTON
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：
2021-01-01 至 2023-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10323278
关键词：
Address Adenocarcinoma Aerobic Alveolar Area Biological Assay Biology Cancer Etiology Cell Respiration Cell Survival Cells Cessation of life Copy Number Polymorphism Data Development Diagnosis Diagnostic tests Dimerization Early treatment Environment Enzymes Epithelial Epithelial Cells Event FGFR1 gene Future Genetic Genetic Models Genus Hippocampus Glycolysis Goals Grant Head and neck structure Health Histologic Human Immunohistochemistry In Vitro Incidence Investigation KRAS2 gene KRASG12D Knowledge Laboratories Life Lung Lung Adenocarcinoma Lung Adenoma Malignant Neoplasms Malignant neoplasm of lung Metabolic Metabolic Pathway Metabolism Mission Modeling Modification Molecular Mus Mutation Neoplasms Non-Small-Cell Lung Carcinoma Nuclear Nuclear Translocation Oncogenes Oncogenic Oxygen Consumption Pathway interactions Patient-Focused Outcomes Patients Phosphorylation Phosphotransferases Post-Translational Protein Processing Production Proteomics Public Health Publications Published Comment Publishing Pyruvate Pyruvate Kinase Research Role Signal Pathway Squamous cell carcinoma Structure of parenchyma of lung TP53 gene Testing The Cancer Genome Atlas Therapeutic Tissues Transcription Coactivator Transgenic Model Type II Epithelial Receptor Cell United States National Institutes of Health adenoma alveolar epithelium base cancer invasiveness cell transformation experimental study glucose metabolism human data human model improved in vivo knock-down member metabolomics mouse model mutant novel programs public health relevance response targeted treatment tumor

项目摘要

PROJECT SUMMARY/ABSTRACT Non-small cell lung cancer (NSCLC) remains the most commonly diagnosed malignancy and leading cause of cancer related deaths worldwide; yet there is a fundamental gap in understanding the biology of lung cancer formation. Sequencing data of human NSCLC revealed KRAS is one of the most frequent oncogene aberrations in lung adenocarcinoma patients. We have recently generated a novel mouse model of NSCLC driven by an oncogenic mutation in KRASG12D and concurrent genetic deletion of MST1/2. When mice express mutant KRASG12D alone in alveolar epithelial type II cells (AECII), we observe focal lung adenomas and neoplasia; however, when MST1/2 are genetically deleted in combination with KRASG12D, we observe aggressive lung adenocarcinoma. To define early mechanisms by which loss of MST1/2 accelerates KRASG12D driven NSCLC, we performed a proteomics screen prior to observing NSCLC on lung tissue from our mouse models. Our proteomic analysis revealed pyruvate kinase M2 (PKM2), a rate-limiting enzyme during glycolysis that catalyzes the production of pyruvate and ATP, is highly expressed, even before the onset of tumors. As improved targeted therapies and diagnostic tests have the potential to significantly improve patient outcomes, we now want to study the mechanism by which loss of MST1/2 promotes increased PKM2. Our central hypothesis is reduced expression of MST1/2 is important in the development of NSCLC. We predict loss of MST1/2 promotes increased abundance and nuclear localization of PKM2, to promote altered glucose metabolism and survival. We plan to test our overall hypothesis by pursuing the following specific aims: Aim 1: To test the hypothesis that MST1 restrains aerobic glucose metabolism in KRAS mutant epithelial cells. Rationale: We have generated new preliminary data that inhibition of MST1 increases PKM2 and pPKM2 in cultured lung epithelial cells expressing KRASG12D. We also have new data using Seahorse assays that inhibition of MST1 significantly increases oxygen consumption rate and ATP production in cultured human NSCLC. We now want to conduct in vivo and in vitro metabolomic analysis to characterize MST1-dependent metabolic programs in human and mouse models driven by mutant KRAS. Aim 2: To test the hypothesis that YAP/TAZ are required for mutant KRAS induction of PKM2. Rationale: We have generated preliminary data using transient knockdown of PKM2 which revealed decreased metabolic activity and proliferation in human NSCLC. In this aim, we will determine what molecular events are causing posttranslational modifications on PKM2 to promote dimerization and nuclear translocation. Using genetic knockdown, we will determine if YAP or TAZ are required for increased expression of PKM2. We will use Seahorse analysis and mass spec to study PKM2 modifications and other metabolic alterations in YAP or TAZ deficient cells. In addition, in response to reviewer comments, we are crossing PKM2fl/fl mice into our genetic model to define the requirement for PKM2 in adenoma-adenocarcinoma development in vivo.

项目概要/摘要非小细胞肺癌（NSCLC）仍然是最常见的恶性肿瘤，也是导致肺癌的主要原因全球癌症相关死亡；然而，对肺癌生物学的理解仍存在根本性差距形成。人类 NSCLC 的测序数据显示 KRAS 是最常见的癌基因畸变之一在肺腺癌患者中。我们最近生成了一种新型 NSCLC 小鼠模型，该模型由 KRASG12D 的致癌突变和同时发生的 MST1/2 基因缺失。当小鼠表达突变体时 KRASG12D单独在肺泡上皮II型细胞（AECII）中，我们观察到局灶性肺腺瘤和肿瘤；然而，当 MST1/2 与 KRASG12D 组合被基因删除时，我们观察到侵袭性肺腺癌。为了确定 MST1/2 缺失加速 KRASG12D 驱动的 NSCLC 的早期机制，在观察小鼠模型肺组织中的非小细胞肺癌之前，我们进行了蛋白质组学筛选。我们的蛋白质组分析揭示了丙酮酸激酶 M2 (PKM2)，它是糖酵解过程中的一种限速酶，可催化丙酮酸和 ATP 的产生，甚至在肿瘤发生之前就高度表达。随着改进的目标疗法和诊断测试有可能显着改善患者的治疗结果，我们现在希望研究 MST1/2 缺失促进 PKM2 增加的机制。我们的中心假设被简化了 MST1/2的表达在NSCLC的发展中很重要。我们预测 MST1/2 的丢失会促进增加 PKM2 的丰度和核定位，促进葡萄糖代谢和存活的改变。我们计划通过追求以下具体目标来检验我们的总体假设：目标 1：检验假设 MST1 抑制 KRAS 突变上皮细胞的有氧葡萄糖代谢。理由：我们有生成了新的初步数据，表明抑制 MST1 会增加培养的肺上皮细胞中的 PKM2 和 pPKM2 表达 KRASG12D 的细胞。我们还使用 Seahorse 检测获得了显着抑制 MST1 的新数据增加培养的人类 NSCLC 中的耗氧率和 ATP 产量。我们现在想要进行体内和体外代谢组学分析，用于表征人类和动物中 MST1 依赖性代谢程序的特征由突变 KRAS 驱动的小鼠模型。目标 2：检验需要 YAP/TAZ 的假设 PKM2 的突变 KRAS 诱导。理由：我们使用瞬态击倒生成了初步数据 PKM2 的研究揭示了人类 NSCLC 中代谢活性和增殖的降低。为了这个目标，我们将确定哪些分子事件导致 PKM2 翻译后修饰以促进二聚化和核易位。使用基因敲低，我们将确定是否需要 YAP 或 TAZ 来增加 PKM2 的表达。我们将使用 Seahorse 分析和质谱来研究 PKM2 修饰和其他 YAP 或 TAZ 缺陷细胞的代谢改变。此外，为了回应审稿人的意见，我们将 PKM2fl/fl 小鼠引入我们的遗传模型，以确定腺瘤-腺癌中对 PKM2 的需求体内发育。