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

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

摘要肾透明细胞癌(Ccrcc)是一种常见的恶性肾癌，具有糖酵解表型。 与VHL抑癌基因的缺失和HIF转录因子的激活有关。高达50%的患者复发 在手术切除后五年内。因此，在临床上迫切需要了解分子。 导致慢性肾细胞癌复发和进展的机制。慢性肾细胞癌的致病机制 进步是一个关键的知识鸿沟。我们最近的发现表明，铜在 先进的ccrcc。在这项提议中，我们询问了铜依赖的代谢重新编程的影响 推动ccrccc进步。铜是酶的金属辅因子，包括细胞色素c氧化酶。 线粒体呼吸必需的络合物(CuCOX)和细胞增殖的金属变构调节剂 和生存。使用患者的原发ccRCC，我们发现铜的积累和CuCOX的强烈增加 与晚期肾细胞癌和复发相关。在细胞系异种移植模型中，功能证据表明 饮食中的铜可促进肿瘤生长，并刺激癌细胞中CuCOX的形成。我们发现，铜 增强电子转移链(ETC)的活性，具有重要的功能后果。高铜诱导(I) 与调节亚基COX7A2L相关的呼吸超复合体(RSC)的组装和(Ii) 线粒体内膜心磷脂(CLS)的积聚和重塑 对于ETC活动是必要的。令人惊讶的是，尽管有糖酵解表型，但铜重新编程的细胞可以恢复 线粒体呼吸，变得高度依赖葡萄糖和CuCOX的活性，因此 对CuCOX抑制剂高度敏感，包括硫化氢(H2S)。重要的是，我们发现 巨噬细胞吞噬的胞内过程是肾癌细胞摄取铜的主要原因。 作为硫化氢供体的小分子正在进行临床前试验，而MP的抑制剂是为 药物疗法，促使将这些途径作为治疗干预的目标进行调查。我们 提示肾癌细胞中铜的慢性蓄积通过TCA循环和ETC活性促进葡萄糖氧化 引起葡萄糖营养不良，刺激肿瘤生长所需的生物能量和生物合成。要确定 铜的来源，我们将研究大吞噬作用，这是一个鲜为人知但却是铜摄取的主要机制 及其在铜向CuCOX分配中的作用(目标1)。为了了解铜对ETC活性的依赖调节，我们 将研究RSC的形成、心磷脂合成和葡萄糖碳通量的机制(目标2)。 最后，由于铜的重新编程对CuCOX抑制剂产生了新的功能漏洞，我们将调查 硫化氢在细胞死亡和肿瘤形成中的作用(目标3)。我们将使用几个模型系统，包括RCC 细胞、体外肿瘤碎片和患者来源的肿瘤移植物。这项研究将通过以下方式确定基本机制 癌细胞中高水平的铜调节其代谢，支持肿瘤的生长。这项研究将引领 对癌症新生物标记物和治疗方法的开发