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 严重脓毒症是一种综合征，估计影响美国 75 万人，其中约 1,900 万人患有严重脓毒症。 每年全世界的人们。随着发病率的上升，死亡率超过20%，并造成严重的死亡 在幸存者中，脓毒症如今被认为是一个公共卫生问题。尽管死亡率一直是 脓毒症导致多器官功能障碍的机制与增加的器官功能障碍密切相关 功能障碍尚未得到很好的了解，因此治疗仍然是反应性的而不是预防性的，并且是非 具体的。最近的证据挑战了之前对脓毒症引起的器官功能障碍的理解： 是由于血流减少引起的细胞死亡，例如急性肾损伤（AKI） 发生在肾血流量正常或增加的情况下；并且它的特点不是急性管状 坏死或凋亡，而是由肾小管上皮细胞 (TEC) 氧化的斑片状、异质区域引起 压力和能量消耗。这种细胞凋亡和坏死的缺乏，以及代谢重新认识 对炎症损伤的反应不仅可以限制急性期的细胞死亡，还可以重新编程能量 决定上皮细胞未来反应的调节途径表明，对这些机制的探索 anisms 代表了潜在的治疗机会。因此，本提案的目标是确定 上皮细胞重新编程代谢以适应炎症损伤和不足的机制 承受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 将在两个具体目标的框架内利用细胞培养和动物模型制定计划。目标 1 将阻止- 探究 AMPK 在调节 TEC 对脓毒症反应的糖酵解和适应阶段中的作用。目标2 将剖析线粒体自噬作为一种节能反应来限制 TEC 氧化应激和细胞死亡的作用 该研究计划将在职业发展计划的背景下制定，该计划将在 以下几页提供了三个基本领域的支持：指导、课程作业和研究。 该计划在匹兹堡大学提供的独特环境中开发，将允许 首席研究员 (PI) 完成过渡到独立所需的关键培训，重点关注 1. 研究脓毒症引起的器官功能障碍的转化模型系统的设计和开发； 2. 的 上皮细胞能量调节途径 (AMPK)、能量利用和周转以及线粒体的量化 败血症期间的软骨质量控制流程； 3. 基本荧光和活体显微镜技术- 评估活体动物病理生理过程的技巧。这些实验将设定 未来工作的阶段，以描述能量调节与器官功能障碍之间的具体途径，以及 利用操纵这些途径的可能性来制定诊断、预防和治疗策略 例如，R01 资助的项目。最终，该职业发展计划的执行将获得独特的地位 PI 作为败血症研究的未来领导者，以改善危重患者的护理。 。