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会导致许多疑难杂症，例如神经代谢疾病利氏综合症几乎肯定会导致儿童死亡，并且在晚期还会恶化。帮助满足新的未满足的医疗需求。为了预防这些疾病的发生，Kaeberlein 小组致力于阐明新的治疗方法线粒体疾病进展的机制和有效药理学的开发为此，我们利用 Leigh 的领先哺乳动物模型。缺失电子传递链结构蛋白亚基 NDUFS4 的综合症这些小鼠表现出严重的症状。我的团队最近发现了 FDA 批准的 mTOR。抑制剂雷帕霉素可以严重减缓疾病进展，并将平均寿命延长约 50% 这些小鼠的雷帕霉素还可以延长野生型小鼠的寿命，延缓易患癌症的小鼠的癌症发病，改善阿尔茨海默氏症模型的神经功能，并预防其他衰老特征。重要证据表明，这种小分子重塑了从 NDUFS4- 中分离出的脑组织中的代谢组 KO 小鼠，包括 NAD+ 水平降低以及从糖酵解向氨基酸分解代谢的转变。我的团队最近观察到 mTOR 和蛋白激酶 C (PKC) 通路严重失活通过磷酸化蛋白质组学分析，雷帕霉素处理的 NDUFS4-KO 小鼠的数据揭示了未知的情况。我已经了解了线粒体生理学中 mTORC2 和 PKC 信号通路之间的关系。有证据表明，抑制 PKC 可以延长这些小鼠的寿命，从而确定了其在病理学中的作用该提案将描述这种关系，阐明其机制含义，并发现新的药理学干预措施来预防线粒体疾病进展基于我的初步数据的假设驱动方法，我将阐明钙依赖性的作用。线粒体相关 ER 膜中的信号传导并探讨个体 PKC 在疾病中的重要性总的来说，这些互补的基础科学研究将通过化学和遗传抑制来实现表型。提供对线粒体生物学的更好理解，揭示信号通路在线粒体中的作用疾病，并阐明细胞间通讯的新机制这些目标的实现。甚至可能对预防正常衰老疾病（如阿尔茨海默氏症、癌症、和心脏病。