Probing the Relationship Between Protein Kinase C and mTOR in Mitochondrial Function

探讨蛋白激酶C与mTOR在线粒体功能中的关系

• 批准号: 10062521
• 负责人: Anthony Steven Grillo
• 金额: $ 6.93万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-12-01 至 2021-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10062521
• 关键词: Achievement; Aging; Alzheimer' s Disease; Alzheimer' s disease model; Amino Acids; Animal Model; Ataxia; Attenuated; Basic Science; Biological; Biology; Birth; Ca(2+)-Transporting ATPase; Calcium; Calcium Signaling; Canis familiaris; Cardiomyopathies; Carrier Proteins; Catabolism; Cell Aging; Cell Fractionation; Cells; Cessation of life; Chemicals; Childhood; Communication; Complex; Coupled; Cytosol; Data; Development; Diabetes Mellitus; Disease; Disease Progression; Disorder of neurometabolic regulation; Electron Transport; Exhibits; FDA approved; FRAP1 gene; Fibroblasts; Fluorescence Microscopy; Focus Groups; Functional disorder; Genetic; Glycolysis; Heart Diseases; Homeostasis; ITPR1 gene; Immunoblotting; Immunofluorescence Immunologic; Individual; Inflammation; Intervention; Knock-out; Knockout Mice; Lactic Acidosis; Late-Onset Disorder; Lead; Leigh Disease; Literature; Longevity; Malignant Neoplasms; Mediating; Medical; Membrane; Membrane Biology; Metabolic; Metabolism; Mitochondria; Mitochondrial Diseases; Modeling; Molecular Target; Mus; Mutation; Nature; Nerve Degeneration; Nervous System Physiology; Onset of illness; Oxidative Phosphorylation; PRKCA gene; Pathologic; Pathology; Pathway interactions; Pharmacology; Phenotype; Physiology; Play; Prevalence; Process; Protein Inhibition; Protein Kinase C; Protein Kinase C Inhibitor; Protein Subunits; Proteins; Proteomics; Regulation; Role; Signal Pathway; Signal Transduction; Sirolimus; Structural Protein; Testing; Thapsigargin; Tissues; Transgenic Mice; Veins; Voltage-Dependent Anion Channel; Washington; Wild Type Mouse; base; brain tissue; chemical genetics; disease phenotype; disorder prevention; healthspan; improved; insight; mTOR Inhibitor; mTOR Signaling Pathway; mTOR inhibition; mTOR protein; metabolome; mitochondrial dysfunction; mitochondrial metabolism; mouse model; neurodegenerative phenotype; novel; novel therapeutics; phosphoproteomics; premature; prevent; protein kinase C beta; protein kinase C gamma; ruboxistaurin; small molecule; success; targeted treatment; uptake; virtual

Mitochondrial dysfunction pathologically causes many incurable diseases such as the neurometabolic disease Leigh Syndrome almost certainly resulting in childhood death. It additionally exacerbates most late- onset diseases such as cancer, Alzheimer's, and heart disease. To help fill the unmet medical need for new treatments that prevent the onset of these diseases, the Kaeberlein Group focuses on elucidating novel mechanisms of mitochondrial disease progression and the development of effective pharmacological interventions with specific molecular targets. To do this, we utilize the leading mammalian model of Leigh Syndrome missing the electron transport chain structural protein subunit NDUFS4. These mice exhibit a severe neurodegenerative phenotype and premature death. My group recently discovered the FDA-approved mTOR inhibitor rapamycin can remarkably attenuate disease progression and increase the mean lifespan by ~50% in these mice. Rapamycin also extends lifespan in wild type mice, delays the onset of cancer in cancer-prone mice, improves neurological function in Alzheimer's models, and prevents other hallmarks of aging. We amassed significant evidence that this small molecule remodels the metabolome in brain tissue isolated from NDUFS4- KO mice, including decreased NAD+ levels and a switch from glycolysis to amino acid catabolism. My group recently observed severe deactivation of the mTOR and protein kinase C (PKC) pathways in rapamycin-treated NDUFS4-KO mice by phosphoproteomic analysis. This data revealed an unknown relationship between the mTORC2 and PKC signaling pathways in mitochondrial physiology. I have acquired evidence that inhibition of PKCs extends lifespan in these mice, establishing its role in the pathology of mitochondrial disease. This proposal will characterize this relationship, elucidate its mechanistic implications, and discover new pharmacological interventions to prevent mitochondrial disease progression taking a hypothesis driven approach based on my preliminary data. I will illuminate the role of calcium-dependent signaling in mitochondria-associated ER membranes and probe the importance of individual PKCs in the disease phenotype through chemical and genetic inhibition. Collectively, these complementary basic science studies will provide a better understanding of mitochondrial biology, uncover the role of signaling pathways in mitochondrial disease, and illuminate novel mechanisms of interorganellar communication. The achievement of these aims may even have broad implications in the prevention of diseases of normative aging such as Alzheimer’s, cancer, and heart disease.

线粒体功能障碍在病理上导致许多不治之症，如神经代谢性疾病， 利氏综合症几乎肯定会导致儿童死亡。它还加剧了最晚- 癌症、老年痴呆症和心脏病等疾病。以帮助填补未满足的医疗需求， 预防这些疾病发作的治疗方法，Kaeberlein集团专注于阐明新的 线粒体疾病进展的机制和有效药理学的发展 针对特定分子靶点的干预措施。为了做到这一点，我们利用Leigh的领先哺乳动物模型 电子传递链结构蛋白亚基NDUFS 4缺失综合征。这些小鼠表现出严重的 神经退行性表型和过早死亡。我的团队最近发现了FDA批准的mTOR 抑制剂雷帕霉素可以显着减缓疾病进展，并增加平均寿命约50%， 这些老鼠雷帕霉素还可以延长野生型小鼠的寿命，延缓癌症易感小鼠的癌症发作， 改善阿尔茨海默病模型的神经功能，并防止其他衰老标志。我们收集了 这一小分子重塑从NDUFS分离的脑组织中的代谢组的重要证据4- KO小鼠，包括NAD+水平降低和糖酵解转换为氨基酸催化剂。 我的研究小组最近观察到， 通过磷酸化蛋白质组学分析测定雷帕霉素处理的NDUFS 4-KO小鼠。这些数据揭示了一个未知的 线粒体生理学中mTORC 2和PKC信号通路之间的关系。我已经获得 有证据表明，抑制蛋白激酶C延长了这些小鼠的寿命，确立了其在糖尿病病理学中的作用。 线粒体疾病这一建议将描述这种关系，阐明其机制的影响， 并发现新的药物干预措施，以防止线粒体疾病的进展， 假设驱动的方法基于我的初步数据。我将阐明钙依赖性 在ESTA相关的ER膜的信号传导，并探测在疾病中的单个PKC的重要性， 表型通过化学和遗传抑制。这些互补的基础科学研究将共同 更好地了解线粒体生物学，揭示线粒体信号通路的作用， 疾病，并阐明细胞器间通讯的新机制。这些目标的实现 甚至可能在预防正常衰老疾病如阿尔茨海默氏症，癌症， 和心脏病。