Metabolic effects of cooper in renal cancer

铜在肾癌中的代谢作用

• 批准号: 10792732
• 负责人: Maria F Czyzyk-Krzeska
• 金额: $ 57.85万
• 依托单位: UNIVERSITY OF CINCINNATI
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-20 至 2028-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10792732
• 关键词: Adjuvant Therapy; Anabolism; Automobile Driving; Biochemical; Biological Markers; Biological Models; COX7A2L gene; Carbon; Cardiolipins; Cell Death; Cell Death Induction; Cell Line; Cell Proliferation; Cell Survival; Cells; Chronic; Citric Acid Cycle; Clear cell renal cell carcinoma; Clinical; Coenzymes; Complex; Copper; Coupled; Data; Development; Electron Transport; Enzymes; Event; Excision; Exposure to; Genetic; Glucose; Glycolysis; Growth; Human; Hydrogen Sulfide; Inner mitochondrial membrane; Investigation; Knowledge; Lipids; Malignant - descriptor; Metabolic; Metabolic Diseases; Metabolism; Metals; Mitochondria; Modeling; Molecular; Oncogenic; Operative Surgical Procedures; Pathogenicity; Pathway interactions; Patients; Pharmacotherapy; Phenotype; Phospholipids; Play; Process; Protein Family; Proteins; Regulation; Relapse; Renal Cell Carcinoma; Renal carcinoma; Research; Respiration; Respiratory physiology; Role; Source; System; Testing; Therapeutic; Therapeutic Intervention; Tumor Suppressor Proteins; Work; Xenograft Model; auxotrophy; cancer cell; cancer therapy; clinical application; cytochrome c oxidase; dietary; experimental study; inhibitor; insight; mitochondrial metabolism; novel; oxidation; preclinical trial; pyruvate dehydrogenase; relapse patients; respiratory; small molecule; standard of care; targeted treatment; transcription factor; tumor; tumor growth; tumor progression; uptake; Adjuvant Therapy; Anabolism; Automobile Driving; Biochemical; Biological Markers; Biological Models; COX7A2L gene; Carbon; Cardiolipins; Cell Death; Cell Death Induction; Cell Line; Cell Proliferation; Cell Survival; Cells; Chronic; Citric Acid Cycle; Clear cell renal cell carcinoma; Clinical; Coenzymes; Complex; Copper; Coupled; Data; Development; Electron Transport; Enzymes; Event; Excision; Exposure to; Genetic; Glucose; Glycolysis; Growth; Human; Hydrogen Sulfide; Inner mitochondrial membrane; Investigation; Knowledge; Lipids; Malignant - descriptor; Metabolic; Metabolic Diseases; Metabolism; Metals; Mitochondria; Modeling; Molecular; Oncogenic; Operative Surgical Procedures; Pathogenicity; Pathway interactions; Patients; Pharmacotherapy; Phenotype; Phospholipids; Play; Process; Protein Family; Proteins; Regulation; Relapse; Renal Cell Carcinoma; Renal carcinoma; Research; Respiration; Respiratory physiology; Role; Source; System; Testing; Therapeutic; Therapeutic Intervention; Tumor Suppressor Proteins; Work; Xenograft Model; auxotrophy; cancer cell; cancer therapy; clinical application; cytochrome c oxidase; dietary; experimental study; inhibitor; insight; mitochondrial metabolism; novel; oxidation; preclinical trial; pyruvate dehydrogenase; relapse patients; respiratory; small molecule; standard of care; targeted treatment; transcription factor; tumor; tumor growth; tumor progression; uptake

Clear cell renal cell carcinoma (ccRCC) is a frequent and malignant renal cancer with a glycolytic phenotype due to the loss of VHL tumor suppressor and activation of HIF transcription factors. Up to 50% of patients relapse within five years after surgical resection. Thus, there is an urgent clinical need to understand the molecular mechanisms leading to ccRCC relapse and advancement. Pathogenic mechanisms underlying ccRCC progression represent a key knowledge gap. Our recent discoveries demonstrate a copper accumulation in advanced ccRCC. In this proposal, we interrogate impact for copper-dependent metabolic reprogramming in driving ccRCC progression. Copper (Cu) is a metal cofactor of enzymes, including cytochrome c oxidase complex (CuCOX) essential for mitochondrial respiration, and a metalloallosteric regulator of cell proliferation and survival. Using patients’ primary ccRCCs, we found accumulation of Cu and increased CuCOX strongly correlated with advanced ccRCC and relapse. In cell line xenograft models, functional evidence shows that dietary Cu drives growth of tumors and stimulates formation of CuCOX in cancer cells. We discovered that Cu enhances electron transfer chain (ETC) activity with important functional consequences. High Cu induces (i) assembly of the respiratory supercomplex (RSC) associated with regulatory subunit, COX7A2L, and (ii) accumulation and remodeling of cardiolipins (CLs), phospholipids of the inner mitochondrial membrane necessary for ETC activity. Surprisingly, despite glycolytic phenotype, Cu-reprogrammed cells recover mitochondrial respiration, become hyperdependent on glucose and on the activity of CuCOX, and therefore are highly sensitive to CuCOX inhibitors, including hydrogen sulfide (H2S). Importantly, we discovered that the endocytic process of macropinocytosis is responsible for major proportion of Cu uptake by renal cancer cells. Small molecules that function as donors of H2S are in preclinical trials, while inhibitors of MP are developed for pharmacotherapies, prompting investigation of these pathways as targets for therapeutic interventions. We propose that chronic accumulation of Cu in RCC cells promotes glucose oxidation via TCA cycle and ETC activity causing glucose auxotrophy and stimulating bioenergy and biosynthesis required for tumor growth. To determine the source of Cu, we will investigate macropinocytosis as little understood but major mechanism of Cu uptake and its role in allocation of Cu to CuCOX (Aim 1). To understand Cu-dependent regulation of ETC activity, we will investigate mechanisms of RSC formation, cardiolipin synthesis and the flux of glucose carbon (Aim 2). Finally, because Cu-reprogramming creates new functional vulnerability to CuCOX inhibitors, we will investigate effects of H2S in cell death and tumor formation (Aim 3). We will use several model systems, including RCC cells, ex-vivo tumor fragments and patient derived tumor grafts. The study will identify basic mechanisms by which high level of Cu in cancer cells regulate their metabolism supporting tumor growth. The research will lead to the development of new biomarkers and therapies for cancer

透明细胞肾细胞癌（CCRCC）是一种经常且恶性肾癌，糖酵解表型应得 VHL肿瘤抑制剂的丧失和HIF转录因子的激活。多达50％的患者中继 手术切除后五年内。那是紧急临床需要了解分子 导致CCRC继电器和进步的机制。 CCRCC的致病机制 进步代表关键知识差距。我们最近的发现证明了铜的积累 高级CCRCC。在此提案中，我们询问了对铜依赖性代谢重编程的影响 驱动CCRC的进展。铜（Cu）是酶的金属辅因子，包括细胞色素C氧化酶 线粒体呼吸所必需的复合物（Cucox），并且是细胞增殖的甲基甲基果质调节剂 和生存。使用患者的主要CCRCC，我们发现Cu的积累和Cucox的增加强烈增加 与高级CCRC和继电器相关。在细胞系异种移植模型中，功能证据表明 饮食CU驱动肿瘤的生长并刺激癌细胞中Cucox的形成。我们发现Cu 增强电子传递链（ETC）活性，具有重要的功能后果。高铜诱导（i） 与调节亚基，COX7A2L和（ii）相关的呼吸超复合（RSC）组装 心脂蛋白（CLS）的积累和重塑，线粒体内部膜的磷脂 ETC活动所必需的。令人惊讶的是，任务糖酵解表型，CU编程细胞恢复 线粒体呼吸变得高依赖于葡萄糖和Cucox的活性，因此是 对包括硫化氢（H2S）在内的Cucox抑制剂高度敏感。重要的是，我们发现 大型细胞增多症的内吞过程是肾脏癌细胞摄取Cu的主要比例。 在临床前试验中，充当H2供体的小分子是在MP中开发的，而MP的抑制剂是为 药物疗法，促使对这些途径作为治疗干预措施的靶标进行研究。我们 提议CU在RCC细胞中的慢性积累通过TCA循环促进葡萄糖氧化和ETC活性 引起葡萄糖的合理植物和刺激肿瘤生长所需的生物合成。确定 CU的来源，我们将研究大胞刺病的理解很少，但主要机制 及其在分配Cu为Cucox中的作用（AIM 1）。要了解Cu依赖于ETC活动的调节，我们 将研究RSC形成，心磷脂合成和葡萄糖碳的通量的机制（AIM 2）。 最后，由于CU编程为Cucox抑制剂创造了新的功能脆弱性，因此我们将研究 H2在细胞死亡和肿瘤形成中的影响（AIM 3）。我们将使用多种模型系统，包括RCC 细胞，前病毒肿瘤碎片和患者衍生的肿瘤移植物。该研究将通过 在癌细胞中，高水平的CU调节其代谢支持肿瘤的生长。这项研究将领导 开发新的生物标志物和癌症疗法