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

• 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
• 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

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.

项目概要 细胞代谢失调是癌症的一个常见特征，但越来越多的研究表明 强调了原发性肿瘤和肿瘤细胞内代谢表型的显着差异 转移至远处器官。虽然高度增殖的原发肿瘤细胞经常表现出瓦尔堡效应， 其特点是偏爱有氧糖酵解以产生 ATP、线粒体​​生物发生的基因和 据报道，氧化磷酸化在转移性癌细胞中上调。因此，详细 肺癌中重编程代谢途径的特征可能揭示对抗肺癌的有用靶标 肿瘤进展并提高患者生存率。为了转移，癌细胞必须首先抵抗失巢凋亡： 由于与细胞外基质失去适当接触而引发的细胞凋亡机制。我们实验室有 先前证明线粒体酶谷氨酸脱氢酶 1 (GDH1) 有助于 通过重新激活 AMPK 来调节生物能反应，从而实现失巢凋亡抵抗和转移 LKB1 缺陷型肺癌。然而，其他线粒体酶在失巢凋亡抵抗中的作用仍然存在 不太了解。为了确定对癌细胞失巢凋亡抵抗力重要的​​其他因素，我们进行了一项 针对肺癌细胞中 120 种线粒体酶的无偏 RNAi 筛选，并鉴定了 ATP- 特定的琥珀酰辅酶A合成酶β亚基（SUCLA2）作为可能对癌细胞很重要的因子 ECM 脱离后的存活率。稳定敲低 SUCLA2 可使肺癌细胞系对失巢凋亡敏感 在体外非贴壁条件下培养。对公开数据的生物信息分析表明 较高的肿瘤 SUCLA2 mRNA 表达与较差的患者生存率相关，我们的研究 免疫组织化学染色表明转移性肺癌中 SUCLA2 蛋白水平较高 与匹配的原发肿瘤相比。此外，我们的数据表明 SUCLA2 敲低显着 增加 ECM 分离癌细胞中活性氧 (ROS) 的产生和失巢凋亡敏感性， 补充抗氧化剂 NAC 可以逆转这种情况。在本研究计划的前半部分，我们将 采用多种方法破译 SUCLA2 如何调节细胞氧化还原状态以促进失巢凋亡 耐药性和肿瘤转移。提案的后半部分列出了一个富有成效的博士后计划 奖学金（K00），该奖学金将以申请人的博士前 F99 培训为基础，为他开始职业生涯做好准备 独立癌症研究员。