ER mitochondrial signaling and alcoholic tissue injury

ER线粒体信号传导和酒精组织损伤

• 批准号: 7860497
• 负责人: Gyorgy Hajnoczky
• 金额: $ 99.74万
• 依托单位: THOMAS JEFFERSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7860497
• 关键词: ADD-1 protein; Address; Adenovirus Vector; Affect; Alcohol abuse; Alcohol consumption; Alcoholism; Alcohols; Apoptosis; Area; Brain; Calcium Signaling; Cell Proliferation; Cell membrane; Cell physiology; Cells; Chronic; Communication; Computational Biology; Coupling; Defect; Development; Disease; Disorder by Site; Electroporation; Endoplasmic Reticulum; Ethanol; Ethanol Metabolism; Exhibits; Fatty Liver; Fluorescence Resonance Energy Transfer; Functional disorder; Gene Expression; Genetic; Goals; Heart; Immune system; Impairment; Indium; Injury; Length; Life; Light; Lipids; Liver; Lung; Measurement; Measures; Mediator of activation protein; Metabolic; Methodology; Methods; Mitochondria; Modeling; Molecular Biology; Monitor; Morphology; Necrosis; Organ; Organelles; Oxidation-Reduction; Oxidative Stress; PPAR alpha; Pancreas; Pharmaceutical Preparations; Predisposition; Proteins; Rattus; Reactive Oxygen Species; Regulation; Research; Research Personnel; Resolution; Role; SRE-2 binding protein; Sarcoplasmic Reticulum; Side; Signal Transduction; Skeletal Muscle; Skeletal muscle injury; Stress; Structure; Subcellular Fractions; Subcellular structure; System; Testing; Time; Tissues; Triglycerides; Width; alcohol abuse therapy; alcohol effect; alcohol exposure; biological adaptation to stress; cell injury; cellular imaging; electron tomography; endoplasmic reticulum stress; feeding; fluorescence imaging; functional disability; in vivo; innovation; interdisciplinary approach; mitochondrial dysfunction; multidisciplinary; novel; problem drinker; public health relevance; reconstruction; response; spatial relationship; technology development; tool; treatment strategy

DESCRIPTION (provided by applicant): This proposal brings together a unique multidisciplinary team of leading experts in alcohol studies and real time dynamic fluorescence imaging of cells and subcellular structures in intact tissues, high resolution electron tomography, sophisticated molecular biology and computational biology to address a fundamental question of how a deregulation of local interactions between organelles can result in global cell dysfunction, tissue damage and disease. The study addresses the regulation of the mitochondria-endoplasmic reticulum (ER)/sarcoplasmic reticulum (SR) interface and its relationship to localized Ca2+ signaling, formation of reactive oxygen species (ROS) and ER stress. The central hypothesis to be tested is that chronic alcohol exposure causes impaired ER/SR-mitochondrial structural and functional coupling and thereby increases the susceptibility to cell injury in liver and skeletal muscle. Our group has developed methods to manipulate the ER/SR-mitochondrial interface and has shown that this has consequences for localized Ca2+ signaling. Our first aim is to further develop these new experimental tools to be able a) to manipulate and measure the ER/SR-mitochondrial interface, b) to monitor the impact of controlled mitochondrial localization on local [Ca2+] signals and local ROS formation and c) to assess ER/SR stress responses in real time at the single cell level. This will help us analyze the relationship between ER/SR stress and changes in localized Ca2+ and ROS signaling. We will then test the concept that the effect of chronic alcohol exposure on ER/SR-mitochondrial interactions leads to impairments in ER/SR-mitochondrial structure, calcium signaling and induces ER/SR- mitochondrial stress responses in the liver and skeletal muscle. Finally, we will determine whether the alterations in ER-mitochondrial morphology and function contribute to alcohol-induced metabolic dysfunction and cell injury in vivo. With these approaches, we can answer fundamental questions about the subcellular organization of the ER-mitochondrial network and its disruption by ethanol. These studies will shed new light on the mechanisms by which alcohol abuse can cause tissue damage and will provide unique opportunities for the development of innovative treatment strategies. PUBLIC HEALTH RELEVANCE: Chronic alcoholism is associated with ER stress and mitochondrial dysfunction and changes in ER and mitochondrial morphology in multiple tissues. The ER/SR-mitochondrial junctions are important for the function and structure of both ER/SR and mitochondria and are envisioned as a target and an important mediator of the alcohol's effect on liver and skeletal muscle injury. A multidisciplinary approach utilizing expertise in alcohol studies and real time dynamic fluorescence imaging of cells and subcellular structures in intact tissues, high resolution electron tomography, sophisticated molecular biology and computational biology will help to address a fundamental question of how a deregulation of local interactions between organelles can result in global cell dysfunction, tissue damage and disease.

描述（由申请人提供）：该提案汇集了一个由酒精研究、完整组织中的细胞和亚细胞结构的实时动态荧光成像、高分辨率电子断层扫描、复杂的分子生物学和计算生物学领域的领先专家组成的独特的多学科团队，以解决细胞器之间局部相互作用的放松管制如何导致整体细胞功能障碍、组织损伤和疾病的基本问题。该研究探讨了线粒体-内质网 (ER)/肌浆网 (SR) 界面的调节及其与局部 Ca2+ 信号传导、活性氧 (ROS) 形成和 ER 应激的关系。待检验的中心假设是，长期接触酒精会导致 ER/SR 线粒体结构和功能耦合受损，从而增加肝脏和骨骼肌细胞损伤的易感性。我们的小组开发了操纵 ER/SR 线粒体界面的方法，并表明这对局部 Ca2+ 信号传导有影响。我们的首要目标是进一步开发这些新的实验工具，以便能够 a) 操纵和测量 ER/SR 线粒体界面，b) 监测受控线粒体定位对局部 [Ca2+] 信号和局部 ROS 形成的影响，c) 在单细胞水平实时评估 ER/SR 应激反应。这将帮助我们分析 ER/SR 应激与局部 Ca2+ 和 ROS 信号传导变化之间的关系。然后，我们将测试以下概念：慢性酒精暴露对 ER/SR-线粒体相互作用的影响会导致 ER/SR-线粒体结构、钙信号传导受损，并诱导肝脏和骨骼肌中的 ER/SR-线粒体应激反应。最后，我们将确定内质网线粒体形态和功能的改变是否导致酒精诱导的体内代谢功能障碍和细胞损伤。通过这些方法，我们可以回答有关内质网线粒体网络的亚细胞组织及其被乙醇破坏的基本问题。这些研究将为酒精滥用导致组织损伤的机制提供新的线索，并将为开发创新治疗策略提供独特的机会。 公共卫生相关性：慢性酒精中毒与内质网应激和线粒体功能障碍以及多个组织中内质网和线粒体形态的变化有关。 ER/SR-线粒体连接对于 ER/SR 和线粒体的功能和结构都很重要，并且被设想为酒精对肝脏和骨骼肌损伤影响的目标和重要介质。利用酒精研究方面的专业知识、完整组织中细胞和亚细胞结构的实时动态荧光成像、高分辨率电子断层扫描、复杂的分子生物学和计算生物学的多学科方法将有助于解决细胞器之间局部相互作用的放松管制如何导致整体细胞功能障碍、组织损伤和疾病的基本问题。