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.

项目摘要/摘要非小细胞肺癌（NSCLC）仍然是最常见的恶性肿瘤和主要原因全球与癌症有关的死亡；然而，了解肺癌的生物学存在根本的差距形成。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。我们最近生成了一个由NSCLC的新型鼠标模型 KRASG12D中的致癌突变和MST1/2的并发遗传缺失。当小鼠表达突变体在肺泡上皮II型细胞（AECII）中，仅Krasg12d，我们观察到局灶性肺腺瘤和肿瘤。但是，当MST1/2与Krasg12d结合遗传删除时，我们观察到侵略性的肺腺癌。为了定义早期机制，MST1/2的损失会加速KRASG12D驱动的NSCLC，在从我们的小鼠模型上观察到肺组织上的NSCLC之前，我们进行了蛋白质组学筛选。我们的蛋白质组学分析表明，丙酮酸激酶M2（PKM2）是糖酵解过程中一种限制酶，催化丙酮酸和ATP的产生甚至在肿瘤发作之前也得到了高度表达。随着目标的改进疗法和诊断测试有可能显着改善患者的预后，我们现在想研究MST1/2损失促进PKM2增加的机制。我们的中心假设减少了 MST1/2的表达在NSCLC的发展中很重要。我们预测MST1/2的丧失会促进 PKM2的丰度和核位置增加，以促进葡萄糖代谢和生存改变。我们计划通过追求以下特定目的来检验我们的整体假设：目标1：检验假设 MST1限制了KRAS突变上皮细胞中的有氧葡萄糖代谢。理由：我们有生成的新初步数据抑制MST1会增加培养的肺上皮的PKM2和PPKM2 表达krasg12d的细胞。我们还使用Seahorse分析有明显抑制MST1的新数据在培养的人NSCLC中增加氧气消耗率和ATP产生。我们现在想进行体内和体外代谢组学分析，以表征人类和人类中MST1的代谢程序小鼠模型由突变体KRAS驱动。目的2：要测试YAP/TAZ需要的假设突变的KRAS诱导PKM2。理由：我们已经使用瞬态敲低生成了初步数据 PKM2显示人类NSCLC的代谢活性和增殖降低。在这个目标中，我们将确定哪些分子事件正在引起对PKM2的翻译后修饰以促进二聚化和核易位。使用遗传敲低，我们将确定是否需要YAP或TAZ增加 PKM2的表达。我们将使用海马分析和质量规格来研究PKM2修改和其他 YAP或TAZ缺陷细胞中的代谢改变。此外，在回应审稿人的评论时，我们是将PKM2FL/FL小鼠跨入我们的遗传模型，以定义腺瘤 - 腺癌中PKM2的需求体内发展。