Regulation of mitochondrial redox homeostasis and signaling in metastatic ovarian cancer

转移性卵巢癌中线粒体氧化还原稳态和信号传导的调节

• 批准号: 10468483
• 负责人: Nadine Hempel
• 金额: $ 24.54万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-08 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10468483
• 关键词: AKT inhibition; Address; Anoikis; Ascites; Biochemical; Cell Adhesion; Cell Culture Techniques; Cell Line; Cell Survival; Cells; Clear Cell; Complement; Data; Deacetylase; Environment; Epigenetic Process; Future; Genetic Transcription; Glutamine; Goals; Greater sac of peritoneum; Homeostasis; Hydrogen Peroxide; Imaging Techniques; Malignant Female Reproductive System Neoplasm; Malignant Neoplasms; Malignant neoplasm of ovary; Mediator of activation protein; Metabolic; Metabolism; Metastatic Malignant Neoplasm to the Ovary; Methylation; Mitochondria; Mitochondrial Proteins; Modeling; Molecular; Monitor; Mus; Neoplasm Metastasis; Nutrient; Ovarian; Oxidants; Oxidation-Reduction; Oxidative Phosphorylation; PTEN gene; Pathway interactions; Patient-Focused Outcomes; Patients; Peritoneal; Phosphoric Monoester Hydrolases; Play; Proteins; Regulation; Reporting; Resistance; Role; SOD2 gene; Signal Pathway; Signal Transduction; Specimen; Stress; Superoxide Dismutase; Superoxides; Survival Rate; Testing; Therapeutic Intervention; Transcriptional Regulation; Tumor-Derived; Up-Regulation; Woman; Work; Xenograft Model; antioxidant enzyme; base; cancer cell; clinically relevant; clinically significant; cytotoxicity; epigenetic regulation; imaging approach; improved; in vivo; inhibitor/antagonist; intraperitoneal; molecular imaging; neoplastic cell; novel; novel therapeutics; oxidation; preservation; promoter; response; sensor; spatiotemporal; therapeutic target; transcription factor; transcriptome sequencing; tumor

Late stage ovarian cancer is marked by poor patient survival and significant metastatic spread throughout the peritoneal cavity. During transcoelomic spread cells must adapt to survive anchorage-independence and to cope with stress associated with matrix detachment and the hostile environment of the ascites. Preliminary findings demonstrate that an important ovarian cancer cell adaptation during anchorage-independence is the up- regulation of two mitochondrial proteins, superoxide dismutase 2 (Sod2), and its regulator, the metabolic and redox sensing deacetylase SIRT3. Moreover, these proteins are necessary for anchorage-independent cell survival and transcoelomic metastasis in vivo. In addition to Sod2’s role as a mitochondrial superoxide scavenger, mechanistic data show that Sod2 shifts the redox landscape of cancer cells to a higher hydrogen peroxide (H2O2) steady-state. This novel non-canonical function of Sod2 as a H2O2 regulator results in mitochondrial redox signaling, as demonstrated by oxidation and inactivation of phosphatases, and enhanced Akt signaling. Preliminary data demonstrate that SIRT3 is an important regulator of Sod2 during anchorage- independence, which points to a novel pro-survival role for SIRT3 during ovarian cancer spread. Thus, the proposal will test the hypothesize that mitochondrial redox signaling is an important regulator of survival adaptations in response to matrix detachment, and that two key mitochondrial proteins, SIRT3 and Sod2, are required for the initiation and regulation of mitochondrial redox signaling in anchorage-independence. This will be addressed using a combination of cell culture models, patient ascites-derived cells, in vivo mouse xenograft models, and molecular, biochemical and imaging techniques to monitor oxidants and redox signaling. Using both unbiased screens and targeted approaches Aim 1 consists of mechanistic studies to delineate how SIRT3/Sod2- regulated mitochondrial H2O2-signaling drives anchorage-independent survival. In Aim 2 the mechanisms of SIRT3 activity and transcriptional regulation will be elucidated how SIRT3 acts as the sensor of matrix detachment in the context of metabolic changes associated with anchorage-independence. In proof-of-principle studies of Aim 3 the vulnerability of Sod2-high tumor cells will be targeted with two approaches based exploiting their sensitivity to H2O2 generating agents and Akt inhibition. Establishing that mitochondrial redox signaling is a necessary adaptation for ovarian cancer anchorage-independent survival and metastasis is crucial in our long- term goal of targeting key metastatic adaptations for novel therapies against ovarian cancer.

晚期卵巢癌的特点是患者生存率低，并且在整个卵巢中有显著的转移性扩散。 腹腔在跨体腔扩散过程中，细胞必须适应锚定独立性科普 与基质脱离和腹水的不利环境相关的应激。初步调查结果 表明在不依赖于锚定的情况下，卵巢癌细胞的一个重要适应是向上， 调节两种线粒体蛋白，超氧化物歧化酶2（SOD 2），及其调节剂，代谢和 氧化还原敏感脱乙酰酶SIRT 3。此外，这些蛋白质是非贴壁依赖性细胞所必需的。 存活和跨体腔转移。除了Sod 2作为线粒体超氧化物的作用外， 清除剂，机械数据显示，Sod 2将癌细胞的氧化还原景观转移到更高的氢 过氧化氢（H2 O2）稳态。Sod 2作为H2 O2调节剂的这种新的非典型功能导致 线粒体氧化还原信号，如磷酸酶的氧化和失活所证明的，并增强 Akt信号。初步数据表明，SIRT 3是锚定过程中Sod 2的重要调节剂。 这表明SIRT 3在卵巢癌扩散期间具有新的促生存作用。因此 这项提案将检验线粒体氧化还原信号是生存的重要调节器的假设 适应响应基质脱离，两个关键的线粒体蛋白，SIRT 3和Sod 2， 所需的启动和调节线粒体氧化还原信号在锚定独立。这将 使用细胞培养模型、患者腹水来源的细胞、体内小鼠异种移植物 模型和分子，生物化学和成像技术，以监测氧化剂和氧化还原信号。同时使用 无偏见的筛选和有针对性的方法目标1包括机制研究，以描述SIRT 3/Sod 2- 受调节的线粒体H2 O2信号传导驱动锚定非依赖性存活。在目标2中， SIRT 3的活性和转录调控将阐明SIRT 3如何作为基质的传感器 在与锚定独立性相关的代谢变化的背景下分离。在原理证明中 目标3的研究中，Sod 2-高肿瘤细胞的脆弱性将通过两种基于开发的方法来靶向 它们对H2 O2产生剂和Akt抑制的敏感性。确定线粒体氧化还原信号是一种 卵巢癌非锚定生存和转移的必要适应在我们的长期研究中至关重要， 长期目标是针对卵巢癌新疗法的关键转移适应。