Nitric oxide and mitochondrial biogenesis in sepsis

脓毒症中的一氧化氮和线粒体生物发生

基本信息

批准号：
8534342
负责人：
CLAUDE A PIANTADOSI
金额：
$ 31.4万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-08-23 至 2014-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8534342
关键词：
5&apos -AMP-activated protein kinase Acute Adenosine Monophosphate Antibiotics Apoptosis Autophagocytosis Biogenesis Biological Models Biology CREB1 gene Caring Cell Death Cell Nucleus Cell Survival Chemical Warfare Chemicals Complex Critical Illness Data Development Failure Genes Genetic Goals Gram-Positive Bacteria Grant Hepatic Hepatocyte Hypoxia Infection Inflammation Inflammatory Knowledge Learning Link Liver Mediating Mediator of activation protein Mitochondria Mitochondrial DNA Multiple Organ Failure Mus NF-kappa B NOS2A gene Nitric Oxide Nitric Oxide Synthase Nucleic Acids Organ Organ failure Organelles Oxidative Phosphorylation Pathway interactions Patients Phenotype Phosphorylation Phosphotransferases Physiological Play Prevalence Prevention Process Production Protein Kinase Proteins Quality Control Receptor Activation Regulation Research Role Sentinel Sepsis Signal Transduction Specificity Standardization Testing Therapeutic Titrations Toll-like receptors Transcriptional Regulation Up-Regulation Work base cost design enzyme activity innovation insight mortality novel novel therapeutic intervention nrf1 protein prevent pro-apoptotic protein programs repaired respiratory response transcription factor

项目摘要

DESCRIPTION (provided by applicant): This revised competitive renewal application seeks to understand how nitric oxide (NO) production in bacterial sepsis causes both mitochondrial damage and regulates mitochondrial quality control, which protects against organ failure. In the previous grant cycle, we discovered that toll-like receptor (TLR) activation of NF-kB- dependent NO synthase (iNOS/NOS2) is involved in the regulation of hepatic mitochondrial biogenesis through the major co-activator, PGC-11 and the adenosine monophosphate (AMP)-activated protein kinase (AMPK). Our preliminary data in mice with sepsis indicate that: NOS2 induction leads to AMPK activation, AMPK helps to activate mitochondrial biogenesis, but not always after ATP depletion, and NOS2 deficiency diminishes AMPK up-regulation and mitochondrial biogenesis and increases apoptosis and inflammation in response to TLR activation. Thus, AMPK appears to activate mitochondrial biogenesis while opposing apoptosis and inflammation in sepsis. In contrast, too much NO is independently associated with mitochondrial damage and loss of NO signal specificity by chemical attack of NO species (NOx) on proteins and nucleic acids. We hypothesize that NOS2, acting in part through AMPK, is required for apposite regulation of the transcriptional program of mitochondrial biogenesis before the failure of ATP production in order to maintain mitochondrial quality control and prevent cell death in sepsis. A test of this hypothesis requires definitions of NO-dependent transcriptional control mechanisms, the role of AMPK, and the chemical biology of mitochondrial NO in relevant model systems. Our approach will focus on the liver as a sentinel organ and on one genetic factor- NOS2- as a quantitative influence on mitochondrial turnover and cell survival. We plan to test these Specific Aims: Aim 1: To determine the role of NOS2 on hepatic mitochondrial biogenesis and cell survival during severe sepsis through quantitative NOS2 gene titration studies. 1A. Define the relationships between NOx-mediated mtDNA and protein damage, respiratory capacity, high-energy metabolite levels, and sepsis-induced hepatic cell death using NOS2 titration. 1B. Assess the importance of NOS2 expression on the regulation of mitochondrial biogenesis in sepsis through CREB and/or NRF-1 induction and regulation of NRF-21 (GABPA) and/or PGC-11 expression. Aim 2: To understand hepatic AMPK activation in sepsis in relation to NOS2, the transcriptional program of mitochondrial biogenesis, and prevention of apoptosis. 2A. Determine if NO-dependent AMPK activation promotes mitochondrial biogenesis in sepsis through CREB and/or prevents apoptosis by inhibitory phosphorylation of pro-apoptotic proteins, Bad and BNIP3. 2B. Determine if pharmacological activation of AMPK in sepsis can promote mitochondrial biogenesis and/or inhibit apoptosis independently of NOS2. These studies will provide new insights into NOx-induced mitochondrial damage in sepsis in the context of physiological mechanisms by which NOS2 regulates mitochondrial biogenesis and the extent to which it is orchestrated by AMPK. By implication, NO regulation of AMPK would play a critical salvage role in MODS, and this knowledge would allow rational new pharmacological approaches to support mitochondrial function while minimizing collateral damage by NOx.

描述（由申请人提供）：这种经过修订的竞争性更新应用程序旨在了解细菌败血症中一氧化氮（NO）的产生如何导致线粒体损伤并调节线粒体质量控制，从而防止器官衰竭。在上一个赠款周期中，我们发现NF-KB-依赖性NO合酶（Inos/NOS2）激活TOLL样受体（TLR）参与了通过主要共振活剂PGC-11的肝线粒体生物发生的调节，PGC-11和腺苷（Amp）磷酸腺苷磷酸盐（AMP）活性蛋白Kinase（Ampactate protein Kinase（Amp）。 Our preliminary data in mice with sepsis indicate that: NOS2 induction leads to AMPK activation, AMPK helps to activate mitochondrial biogenesis, but not always after ATP depletion, and NOS2 deficiency diminishes AMPK up-regulation and mitochondrial biogenesis and increases apoptosis and inflammation in response to TLR activation.因此，AMPK似乎激活了线粒体生物发生，同时反对败血症的凋亡和炎症。相比之下，太多的否与线粒体损伤和无信号特异性的丧失无独立的蛋白质和核酸上的非信号特异性相关。我们假设NOS2部分通过AMPK起作用，是对ATP产生之前的线粒体生物发生转录程序的适当调节，以维持线粒体质量控制并防止脓毒症的细胞死亡。对该假设的检验需要定义NO依赖性的转录控制机制，AMPK的作用以及线粒体NO在相关模型系统中的化学生物学。我们的方法将集中于肝脏作为前哨器官，而将一种遗传因子NOS2-作为对线粒体周转和细胞存活的定量影响。我们计划测试这些特定目标：目标1：通过定量的NOS2基因滴定研究，在严重败血症期间，在严重败血症期间，NOS2对肝脏线粒体生物发生和细胞存活的作用。 1a。定义NOX介导的mtDNA与蛋白质损伤，呼吸能力，高能代谢物水平以及使用NOS2滴定对肝诱导的肝细胞死亡之间的关系。 1B。评估NOS2表达对通过CREB和/或NRF-1诱导以及NRF-21（GABPA）（GABPA）和/或PGC-11表达的调节的NOS2表达对线粒体生物发生的调节的重要性。目的2：了解败血症中的肝AMPK激活NOS2，线粒体生物发生的转录程序和预防凋亡。 2a。确定非依赖性AMPK激活是否通过CREB促进败血症中的线粒体生物发生和/或通过抑制促凋亡蛋白的抑制性磷酸化（BAD和BNIP3）来预防凋亡。 2b。确定AMPK在败血症中的药理激活是否可以促进线粒体生物发生和/或独立于NOS2抑制凋亡。这些研究将在生理机制中对NOS2调节线粒体生物发生及其由AMPK策划的程度的生理机制，对败血症中NOX诱导的线粒体损害的新见解。暗示，AMPK的任何调节都不会在mod中发挥关键的打捞作用，并且这些知识将允许合理的新药理方法支持线粒体功能，同时最大程度地减少NOX的侧支损害。