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

基本信息

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
项目类别：
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&apos s Disease Alzheimer&apos 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

项目摘要

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引起许多无法治愈的疾病，例如神经代谢疾病利综合症几乎肯定会导致儿童死亡。此外，它最晚期加剧了癌症，阿尔茨海默氏病和心脏病等发作疾病。帮助满足未满足的新需求阻止这些疾病发作的治疗方法，Kaeberlein组专注于阐明新颖的新型线粒体疾病进展的机制和有效药物的发展特定分子靶标的干预措施。为此，我们利用了Leigh的领先哺乳动物模型综合征缺少电子传输链结构蛋白亚基NDUFS4。这些小鼠暴露了严重的神经退行性表型和过早死亡。我的小组最近发现了FDA批准的MTOR 抑制剂雷帕霉素可以显着减轻疾病的进展，并使平均寿命增加约50％这些老鼠。雷帕霉素还延长了野生型小鼠的寿命，延迟了容易发生癌症小鼠的癌症发作，改善阿尔茨海默氏症模型中的神经功能，并防止其他衰老的标志。我们积累了大量证据表明，该小分子重塑了从NDUFS4-分离的脑组织中的代谢组 KO小鼠，包括NAD+水平降低和从糖酵解转变为氨基酸分解代谢。我小组最近观察到MTOR和蛋白激酶C（PKC）途径的严重失活雷帕霉素处理的NDUFS4-KO小鼠通过磷酸蛋白质组学分析。这些数据揭示了一个未知的线粒体生理学中MTORC2和PKC信号通路之间的关系。我已经获得了抑制PKC在这些小鼠中延长寿命的证据，确定了其在病理中的作用线粒体疾病。该建议将表征这种关系，阐明其机械含义，并发现新的药物干预措施，以防止线粒体疾病进展假设驱动的方法基于我的初步数据。我将阐明依赖钙的作用线粒体相关的ER膜中的信号传导，并探测单个PKC在该疾病中的重要性通过化学和遗传抑制的表型。总的来说，这些完整的基础科学研究将提供更好地了解线粒体生物学，揭示信号通路在线粒体中的作用疾病，并阐明了组织间通信的新型机制。这些目标的成就甚至可能对预防正常衰老的疾病（例如阿尔茨海默氏症，癌症）有广泛的影响和心脏病。