The role of energy regulation in the epithelial cell response to sepsis and the origin of multiple organ dysfuntion

能量调节在上皮细胞对脓毒症反应中的作用和多器官功能障碍的起源

• 批准号: 9321179
• 负责人: Hernando Gomez Danies
• 金额: $ 18.65万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-08-01 至 2019-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9321179
• 关键词: Acute; Acute Kidney Tubular Necrosis; Acute Renal Failure with Renal Papillary Necrosis; Adenosine Monophosphate; Adenosine Triphosphate; Affect; Animal Model; Animals; Apoptosis; Area; Autophagocytosis; Biological Models; Blood; Blood flow; Cell Culture Techniques; Cell Death; Cell Survival; Cells; Complex; Critical Care; Critical Illness; Cyclic AMP-Dependent Protein Kinases; Cytosol; Data; Death Rate; Defense Mechanisms; Development; Development Plans; Diagnostic; Environment; Epithelial; Epithelial Cells; FRAP1 gene; Fluorescence; Functional disorder; Funding; Future; Genus Hippocampus; Glucose; Glycolysis; Glycolysis Inhibition; Goals; Human; Incidence; Inflammation; Inflammatory; Injury; Kidney; Knock-out; Knowledge; Ligation; Link; Mediating; Mentors; Metabolic; Metabolism; Mitochondria; Modeling; Modification; Morbidity - disease rate; Mus; Necrosis; Nicotinamide adenine dinucleotide; Organ; Oxidative Phosphorylation; Oxidative Stress; Oxygen Consumption; Pathway interactions; Patients; Pharmacology; Phase; Phosphorylation; Positioning Attribute; Preventive; Principal Investigator; Process; Production; Program Development; Proliferating; Protein Kinase; Proteins; Public Health; Puncture procedure; Quality Control; Reactive Oxygen Species; Regulation; Regulatory Pathway; Renal Blood Flow; Research; Respiration; Rodent Model; Role; SIRT1 gene; Sepsis; Signal Transduction; Small Interfering RNA; Source; Survivors; Syndrome; Techniques; Therapeutic; Tissues; Training; Tubular formation; United States; Universities; Warburg Effect; Work; base; career development; cell injury; design; energy balance; experimental study; extracellular; hypoxia inducible factor 1; improved; intravital microscopy; knock-down; liver metabolism; mortality; oxidation; oxidative damage; programs; response; tumor

PROJECT ABSTRACT Severe sepsis is a syndrome estimated to affect 750,000 people in the United States and about 19 million people worldwide every year. With a rising incidence, death rates exceeding 20% and causing significant mor- bidity in survivors, sepsis is considered today a public health problem. Despite that mortality has been consist- ently associated with increasing organ dysfunction, the mechanisms by which sepsis causes multiple organ dysfunction are not well understood, and hence therapy remains reactive rather than preventive, and non- specific. Recent evidence has challenged the previous understanding of sepsis-induced organ dysfunction as being due to decreased blood flow-induced cell death by showing for example that acute kidney injury (AKI) occurs in the setting of normal or increased renal blood flow; and that it is characterized not by acute tubular necrosis or apoptosis, but rather by patchy, heterogeneous areas of tubular epithelial cell (TEC) oxidative stress and energy depletion. This paucity of apoptosis and necrosis, and the recognition that metabolic re- sponses to inflammatory injury may not only limit cell death in the acute phase but also, re-program energy regulatory pathways to determine future responses of epithelial cells, suggests that exploration of these mech- anisms represents potential therapeutic opportunities. Accordingly, the goal of this proposal is to determine the mechanisms by which the epithelial cell re-programs metabolism to adapt to inflammatory injury, and to under- stand the impact of these modifications on cell and organ function, and cell survival. The proposed research plan will be developed using cell culture and animal models in the frame of two specific aims. Aim 1 will deter- mine the role of AMPK in regulating the glycolytic and adaptive phases of the TEC response to sepsis. Aim 2 will dissect the role of mitophagy as an energy conserving response to limit TEC oxidative stress and cell death This research program will be framed in the context of a career development plan that will be described in the following pages and that is supported on three fundamental domains: Mentoring, Coursework and Research. The development of this program in the unique environment provided by the University of Pittsburgh, will allow the principal investigator (PI) to complete key training necessary to transition to independence focused on 1. The design and development of translational model systems to study sepsis induced organ dysfunction; 2. The quantification of epithelial cell energy regulatory pathways (AMPK), energy utilization and turnover, and mito- chondrial quality control processes during sepsis; and 3. Basic fluorescence and intravital microscopy tech- niques for the assessment of pathophysiologic processes in the living animal. These experiments will set the stage for future work to characterize the specific pathways linking energy regulation to organ dysfunction, and harness the possibility of manipulating these pathways to develop diagnostic, preventive and therapeutic strat- egies in a R01-funded project. Ultimately, the execution of this career development plan will uniquely position the PI as a future leader in sepsis research to improve the care of the critically ill patient. .

项目摘要 严重败血症是一种综合征，估计会影响美国750,000人，约1900万 每年都在全球。随着发病率的上升，死亡率超过20％ 幸存者的婚姻，败血症被认为是一个公共卫生问题。尽管死亡率已经组成 - 与增加器官功能障碍相关，败血症引起多器官的机制 功能障碍尚不清楚，因此治疗仍然是反应性的，而不是预防性的，非 - 具体的。最近的证据挑战了先前对败血症引起的器官功能障碍的理解 由于急性肾脏损伤（AKI），由于血液流动诱导的细胞死亡减少所致 发生在正常或增加肾血流的情况下；并且它的特征不是急性管状 坏死或细胞凋亡，而是通过管状上皮细胞（TEC）氧化的片状，异质区域 压力和能量消耗。凋亡和坏死的这种稀少，以及代谢恢复的认识 发作性损伤的发起不仅可能限制急性期细胞死亡，而且还可以重新编程能量 确定上皮细胞未来反应的调节途径表明，这些机甲的探索 Anisms代表潜在的治疗机会。因此，该提议的目的是确定 上皮细胞重新编程代谢以适应炎症性损伤的机制，并 具有这些修饰对细胞和器官功能以及细胞存活的影响。拟议的研究 计划将使用两个特定目的的细胞培养和动物模型制定。 AIM 1将阻止 - 挖掘AMPK在调节TEC对败血症反应的糖酵解和适应阶段中的作用。目标2 将剖析线粒体的作用，作为限制TEC氧化应激和细胞死亡的能量响应的作用 该研究计划将在职业发展计划的背景下进行构建，该计划将在 遵循页面，并在三个基本领域得到支持：指导，课程和研究。 在匹兹堡大学提供的独特环境中，该计划的制定将允许 首席研究员（PI）要完成过渡到独立性所必需的关键培训。 研究和开发败血症诱导器官功能障碍的翻译模型系统； 2 定量上皮细胞能量调节途径（AMPK），能量利用和周转，以及MITO- 败血症期间的软骨质量控制过程； 3。基本荧光和插入显微镜技术 - 评估活动物中病理生理过程的有益机。这些实验将设置 未来工作的阶段，以表征将能量调节与器官功能障碍联系起来的特定途径，并且 利用操纵这些途径以发展诊断，预防和治疗性层的可能性 R01资助的项目中的egies。最终，该职业发展计划的执行将唯一地位 PI是败血症研究的未来领导者，以改善对重症患者的护理。 。