Respiration in Sepsis

脓毒症时的呼吸

基本信息

批准号：
8666533
负责人：
CLAUDE A PIANTADOSI
金额：
--
依托单位：
DURHAM VA MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-01-01 至 2016-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8666533
关键词：
5&apos -AMP-activated protein kinase AMP-activated protein kinase kinase Adenosine Monophosphate Aging Agonist Animal Model Antibiotics Apoptosis Apoptotic Area Autophagocytosis Basic Science Biogenesis CREB1 gene Carbon Monoxide Caring Cause of Death Cell Death Cell Survival Cell model Chemical Warfare Data Disease Energy Metabolism Event Failure Funding Gene Expression Regulation Genes Genetic Genetic Programming Goals Health Hepatic Hepatocyte High Prevalence Hypoxia Immune system Infection Inflammation Inflammatory Lead Life Link Liver Mediating Mediator of activation protein Mitochondria Mitochondrial DNA Mitochondrial Proteins Molecular Multiple Organ Failure Mus Nitric Oxide Nitric Oxide Synthase Organ Oxidation-Reduction Pathogenesis Pathway interactions Phosphorylation Prevention Process Production Protein Kinase Proteins Quality Control Receptor Activation Regulation Respiration Role Sentinel Sepsis Signal Transduction Specificity Standardization Staphylococcus aureus Syndrome Testing Therapeutic Thinking Tissues Titrations Toll-like receptors Transcription factor genes Transcriptional Regulation Up-Regulation Veterans Work cell injury cost design high throughput screening innovation insight mortality novel nrf1 protein prevent programs public health relevance respiratory response transcription factor

项目摘要

DESCRIPTION (provided by applicant): This VA Merit Review renewal application proposes to study transcriptional regulation of the pro-survival program of mitochondrial biogenesis during experimental S. aureus sepsis. The need for basic research in this area is high due to the high prevalence and cost of sepsis-induced multiple organ dysfunction syndrome (MODS) in older Veterans. This high mortality syndrome is due in part to mitochondrial damage, which is mechanistically poorly understood. Using earlier funding, we discovered that mitochondrial biogenesis in sepsis is activated before energy crisis, while a rescue pathway is delayed involving the adenosine monophosphate (AMP)-activated protein kinase (AMPK) and PGC-1¿ co-activator. New preliminary data links AMPK activation to nitric oxide synthase-2 (NOS2) induction in sepsis, does not require high AMP/ATP, and loss of AMPK activation in nitric oxide synthase-2 (NOS2) deficient mice is accompanied by worsening inflammation. AMPK may serve as an activator of three crucial transcription factors- CREB, NRF-1, and NRF- 2 (GABPA) for mitochondrial biogenesis in sepsis, but NO regulation also induces mitochondrial damage by loss of NO signal specificity and indiscriminate chemical attack on mitochondria by NO species (NOx). We propose that AMPK activation by impending energy failure up-regulates the mitochondrial damage control program in sepsis and if so, NO-independent pharmacological AMPK activation could control excess NO production and mitigate sepsis-induced MODS. We will focus on AMPK in the liver, a sentinel organ, and NOS2 as a quantitative influence on mitochondrial turnover and apoptosis using pharmacological AMPK activation to limit NO-induced mitochondrial damage. Our Specific Aims are: Aim 1: To understand hepatic AMPK activation in sepsis in relation to NOS2 induction, the program of mitochondrial biogenesis, and the prevention of apoptosis. 1A. Define the relationships between AMPK activation and NOx-mediated mtDNA and protein damage, respiratory capacity, high-energy metabolites, and sepsis-induced hepatic cell death using NOS2 gene titration. 1B. Determine if AMPK activation through NO-dependent CREB/NRF-1 activity promotes mitochondrial biogenesis and inhibitory phosphorylation of pro-apoptotic Bad and BNIP3, preventing apoptosis in sepsis. Aim 2: To determine if specific strategies to activate or inhibit AMPK independently of NOS2 regulate hepatic mitochondrial biogenesis and cell survival and lessen NO-dependent cell damage in sepsis. 2A. Determine if the pharmacological activation of AMPK independently of NOS2 promotes mitochondrial biogenesis and/or inhibits apoptosis in sepsis. We will also use high-throughput screening to identify one or more novel selective agonists of AMPK. 2B. Compare NO-dependent and NO-independent AMPK activation for effects on the transcriptional regulation of mitochondrial biogenesis in sepsis using CREB and PGC-1¿ as key readouts. These studies will provide new molecular data on AMPK regulation of mitochondrial biogenesis in sepsis, and on the extent to which NO production is regulatory. Proof-of-concept would lead to rational pharmacological approaches to activate and support mitochondrial biogenesis while minimizing NO-induced collateral damage. Understanding these fundamental regulatory mechanisms is needed to design forward-thinking therapeutic approaches to protect mitochondrial quality control during sepsis.

描述（由申请人提供）：这项VA值得回顾的续签应用建议，以研究实验性金黄色葡萄球菌败血症期间线粒体生物发生的促生物生物生物生物的转录调节。由于老年退伍军人中败血症引起的多个器官功能障碍综合征（MOD）的高患病率和成本高，因此对该领域的基础研究的需求很高。这种高死亡率综合征的部分原因是线粒体损害的机械理解不足。使用较早的资金，我们发现败血症中的线粒体生物发生在能量危机前被激活，而救援途径则延迟涉及腺苷单磷酸（AMP）激活的蛋白激酶激酶（AMPK）和PGC-1？co-激活剂。新的初步数据将AMPK激活与脓毒症中的一氧化氮合酶-2（NOS2）诱导，不需要高AMP/ATP，并且通过担心被担心入射来实现一氧化氧化物合酶-2（NOS2）（NOS2）（NOS2）中AMPK激活的损失。 AMPK可以作为三种至关重要的转录因子的激活剂-CREB，NRF-1和NRF-2（GABPA），用于脓毒症中的线粒体生物发生，但没有任何调节也会通过无需任何物种（Nox）（Nox）对线粒体造成无信号特异性和无歧视化学攻击。我们提出，通过迫在眉睫的能量故障来激活AMPK激活败血症中的线粒体损伤控制程序，如果是这样，则无独立的药物AMPK激活可以控制不超过生产，并减轻败血症诱导的mod。我们将使用药物AMPK激活来限制无诱导的线粒体损伤的肝脏，前哨器官和NOS2的AMPK，作为对线粒体周转和凋亡的定量影响。我们的具体目的是：目标1：了解与NOS2诱导，线粒体生物发生程序和预防凋亡有关的败血症中的肝AMPK激活。 1a。定义AMPK激活与NOX介导的mtDNA与蛋白质损伤，呼吸能力，高能量代谢产物以及败血症诱导的肝细胞死亡，使用NOS2基因滴定。 1B。确定通过无依赖性CREB/NRF-1活性激活AMPK是否会促进线粒体的生物发生并抑制促凋亡坏和BNIP3，从而阻止败血症的凋亡。目标2：确定与NOS2无关激活AMPK的特定策略是否调节肝脏线粒体生物发生和细胞存活以及败血症中无依赖性细胞损伤。 2a。确定AMPK的药理学激活是否独立于NOS2促进了线粒体生物发生和/或抑制败血症的凋亡。我们还将使用高通量筛选来识别AMPK的一种或多种新颖的选择性激动剂。 2b。比较使用CREB和PGC-1作为关键读数的无依赖性和无独立的AMPK激活对败血症中线粒体生物发生的转录调节的影响。这些研究将提供有关败血症线粒体生物发生的AMPK调节的新分子数据，以及在没有生产的程度上进行调节。概念证明将导致有理药物的方法激活和支持线粒体生物发生，同时最大程度地减少无诱导的侧支损害。需要了解这些基本的调节机制来设计具有前瞻性的治疗方法，以保护败血症期间的线粒体质量控制。