Metabolic reprogramming and tumor progression in LKB1 deficient NSCLC

LKB1 缺陷型 NSCLC 的代谢重编程和肿瘤进展

基本信息

批准号：
10305441
负责人：
Austin C Boese
金额：
$ 4.75万
依托单位：
EMORY UNIVERSITY
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：
2021-09-01 至 2023-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10305441
关键词：
Adenocarcinoma Anoikis Antioxidants Apoptotic Bioenergetics Biogenesis Bioinformatics Biological Markers Blood Vessels Cancer Etiology Cancer Patient Cancer cell line Cardiovascular system Cell Death Cell Survival Cells Cessation of life Characteristics Clinical Clinical Management Complex Custom Data Diagnosis Disseminated Malignant Neoplasm Distant Drug Targeting Enzymes Equilibrium Exhibits Extracellular Matrix Fellowship Future Genes Genetic Glutamate Dehydrogenase Goals Homeostasis Immune Evasion Immune checkpoint inhibitor Immunohistochemistry Immunotherapy In Vitro Laboratories Large Cell Carcinoma Link Malignant Neoplasms Malignant neoplasm of lung Mediating Metabolic Metabolic Pathway Metabolism Mitochondria Molecular Neoplasm Metastasis Non-Small-Cell Lung Carcinoma Nonmetastatic Nutrient Organ Oxidation-Reduction Oxidative Phosphorylation Oxidative Stress Pathway interactions Patient-Focused Outcomes Patients Phenotype Play Primary Neoplasm Process Production Proteins RNA interference screen Reactive Oxygen Species Reporting Research Research Personnel Research Proposals Resistance Role STK11 gene Signal Transduction Squamous cell carcinoma Stains Succinate-CoA Ligases Supplementation Testing Tissues Training Tumor Escape United States Uric Acid Warburg Effect aerobic glycolysis cancer cell cancer therapy career checkpoint therapy combat experimental study improved in vivo inhibitor/antagonist knock-down lung cancer cell lymphatic vessel mRNA Expression metabolic phenotype metabolic profile metabolomics mortality neoplastic cell novel pre-doctoral preclinical efficacy preference programs response targeted biomarker therapeutic target therapy resistant tumor tumor progression tumor-immune system interactions

项目摘要

PROJECT SUMMARY Dysregulated cellular metabolism is a common characteristic of cancer, but a growing body of research highlights significant differences in metabolic phenotypes both within primary tumors, and in tumor cells that metastasize to distant organs. While highly proliferative primary tumor cells often exhibit the Warburg effect, marked by a preference for aerobic glycolysis to generate ATP, genes for mitochondrial biogenesis and oxidative phosphorylation are reported to be upregulated in metastatic cancer cells. Therefore, detailed characterization of reprogrammed metabolic pathways in lung cancer may reveal useful targets to combat tumor progression and improve patient survival. In order to metastasize, cancer cells must first resist anoikis: an apoptotic cell death mechanism triggered by loss of proper contact with the extracellular matrix. Our lab has previously demonstrated that the mitochondrial enzyme glutamate dehydrogenase 1 (GDH1) contributes to anoikis resistance and metastasis by regulating the bioenergetic response through reactivation of AMPK in LKB1-deficient lung cancer. However, the role of other mitochondrial enzymes in anoikis resistance remains poorly understood. To identify other factors important for cancer cell anoikis resistance, we performed an unbiased RNAi screen targeting 120 mitochondrial enzymes in lung cancer cells and identified the ATP- specific Succinyl-CoA Synthetase beta subunit (SUCLA2) as a factor that may be important for cancer cell survival after ECM detachment. Stable knockdown of SUCLA2 sensitized lung cancer cell lines to anoikis when cultured under non-adherent conditions in vitro. Bioinformatic analysis of publicly available data indicates that higher tumor SUCLA2 mRNA expression is associated with poor patient survival, and our immunohistochemistry staining suggests that SUCLA2 protein levels are higher in metastatic lung cancer compared to matched primary tumors. Additionally, our data suggests that SUCLA2 knockdown significantly increases reactive oxygen species (ROS) production and anoikis sensitivity in ECM detached cancer cells, which can be reversed with antioxidant NAC supplementation. In the first half of this research proposal, we will employ multiple approaches to decipher how SUCLA2 modulates cellular redox status to promote anoikis resistance and tumor metastasis. The second half of the proposal lays out a plan for a productive postdoctoral fellowship (K00) that will build on applicant's predoctoral F99 training and prepare him well to start a career as an independent cancer researcher.

项目摘要细胞代谢失调是癌症的常见特征，但越来越多的研究体系强调了原发性肿瘤内代谢表型的显着差异，以及在肿瘤细胞中转移到遥远的器官。虽然高度增殖的原发性肿瘤细胞经常表现出Warburg效应，但以有氧糖酵解产生ATP的偏爱，线粒体生物发生和据报道，氧化磷酸化在转移性癌细胞中被上调。因此，详细肺癌中重编程代谢途径的表征可能会揭示有用的目标。肿瘤进展并改善患者的生存率。为了转移，癌细胞必须首先抵抗Anoikis：凋亡细胞死亡机制是由于与细胞外基质的正确接触而触发的。我们的实验室有以前证明了线粒体酶谷氨酸脱氢酶1（GDH1）有助于通过调节AMPK在AMPK中的重新激活来调节生物能反应，抵抗和转移 LKB1缺陷型肺癌。但是，其他线粒体酶在抗阳极抗性中的作用仍然存在理解不佳。为了确定对癌细胞抗性抗性重要的其他因素，我们进行了无偏的RNAi筛查靶向120个线粒体酶的肺癌细胞，并鉴定出ATP- 特定的琥珀酰-COA合成酶β亚基（SUCLA2）是可能对癌细胞重要的因素 ECM脱离后的生存。稳定地敲低sucla2敏感的肺癌细胞线对厌氧菌在体外培养在非遵循条件下。对公开数据的生物信息学分析表明较高的肿瘤sucla2 mRNA表达与患者生存不良有关，我们免疫组织化学染色表明，转移性肺癌中的sucla2蛋白水平较高与匹配的原发性肿瘤相比。此外，我们的数据表明Sucla2敲低在ECM脱离的癌细胞中增加活性氧（ROS）的产生和厌氧敏感性，可以通过补充NAC抗氧化剂来逆转。在本研究建议的上半年，我们将采用多种方法来解密Sucla2如何调节细胞氧化还原状态以促进Anoikis 抗性和肿瘤转移。提案的下半部分制定了一个有效的博士后计划奖学金（K00）将在申请人的F99培训前建立，并为他做好准备，以开始职业独立的癌症研究员。