Regulation of Innate Immunity and Inflammation Through Nuclear Reprogramming

通过核重编程调节先天免疫和炎症

• 批准号: 10339416
• 负责人: Jack J Hawiger
• 金额: --
• 依托单位: VETERANS HEALTH ADMINISTRATION
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-01-01 至 2024-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10339416
• 关键词: Age; Anti-Bacterial Agents; Anti-Inflammatory Agents; Antibiotics; Bacteria; Bacterial Infections; Binding; Binding Proteins; Blood; Brain; Carbohydrates; Carnitine; Cell Nucleus; Cells; Cholesterol; Clinical Trials; Complex; Coupled; Elements; Endothelial Cells; Endothelium; Endotoxins; Esters; Fatty Acids; Gene Expression; Genes; Genetic Transcription; Genome; Genomics; Glucose; Goals; Gram-Negative Bacteria; Heart; Hereditary Disease; Human; Hyperglycemia; Hyperlipidemia; Hypersensitivity; Hypertriglyceridemia; Hypotension; Immune; Immunotherapy; Impaired cognition; Importins; Incidence; Individual; Infection; Inflammation; Inflammation Mediators; Inflammatory; Injury; Interleukin-1 beta; Interleukin-6; Kidney; Laboratories; Link; Lipopolysaccharides; Liver; Lung; Mediating; Metabolic; Metabolic Diseases; Metabolic syndrome; Microcirculation; Modeling; Monitor; Multiple Organ Failure; Mus; Natural Immunity; Nuclear Import; Nuclear Pore Complex; Organ; Outcome; Pathway interactions; Patients; Peptides; Peritonitis; Platelet Activation; Population; Production; Proteins; Regulation; Regulatory Element; Reporting; Research; Sepsis; Septic Shock; Shock; Signal Pathway; Spleen; Sterols; Stress; Survival Rate; Survivors; TNF gene; Testing; Thrombocytopenia; Tissues; Treatment Protocols; Triglycerides; Veterans; Virulence Factors; antimicrobial; antimicrobial drug; base; blood glucose regulation; carbohydrate transport; clinically relevant; design; genomic data; glycogenolysis; improved; innovation; lipid biosynthesis; microbial; mortality; multiorgan damage; novel; nuclear reprogramming; nucleocytoplasmic transport; prevent; protein transport; recurrent infection; response; transcription factor; western diet; wounded soldier

PROJECT SUMMARY Regulation of Innate Immunity and Inflammation Through Nuclear Reprogramming We propose to advance our mechanistic studies of innate immunity and inflammation in severe infections that evolve into their end stage, septic shock, thereby threatening the lives of wounded soldiers on the battlefield, our Veterans, and the civilian population. Those Veterans (and non-Veteran patients) between the ages of 65 to 85 are particularly prone to septic shock with a 45% mortality rate while many survivors suffer debilitating cognitive decline. We are pursuing our long-term goal to decode innate immunity and inflammation with a new class of cell-penetrating Nuclear Transport Modifiers (NTMs) that reprogram the inflammatory regulome. One of these anti-inflammatory peptides achieved an unprecedented enhancement of innate immunity-mediated bacterial clearance in the lungs (700-fold), spleen (300-fold), and blood (200-fold), all taking place before antimicrobial therapy. Concurrently, it improved the blood markers of microvascular endothelial injury, including thrombocytopenia. When combined with anti-microbial therapy, NTM increased the survival rate to 55%, compared to a 30% survival rate in the antibiotic-only group. This NTM peptide, cSN50.1, simultaneously targets two nuclear transport shuttles, importins α5 and β1. To unravel the mechanism of innate immunity and inflammation that underlie microbial and metabolic disorders, we designed, produced, and tested two novel mono-selective NTMs: (i) Importin α5-selective cSN50.1α, and (ii) Importin β1-selective cSN50.1β. Strikingly, the importin α5-selective peptide produced an 80% survival rate in endotoxin shock induced by lipopolysaccharide (LPS), one of the most potent virulence factors of Gram-negative bacteria responsible for two thirds of septic shock cases. This new NTM also effectively controlled the Western Diet-induced hypersensitivity to lethal endotoxin shock while preventing liver glycogenolysis, which causes hyperglycemia and hypertriglyceridemia linked to the nuclear transport of Carbohydrate Responsive Elements-Binding Proteins (CHREBPs). In turn, the Imp β1-selective peptide suppressed lipogenesis mediated by the nuclear transport of the Sterol Regulatory Element-Binding Proteins (SREBPs) that control over 30 genes involved in the production of triglycerides, cholesterol, and fatty acids, a hallmark of metabolic syndrome. We hypothesize that the proposed use of selective NTMs with antimicrobial agent(s) would untangle multiorgan damage and mortality in severe infections. Furthermore, we posit that the Importin α5/SRTFs/CHREBPs and Imp β1/SREBPs pathways are interdependent, thereby predisposing individuals with metabolic syndrome to hyperacute responses to severe infections. Hence, in Aim 1, we will investigate the mechanism of the NTM’s beneficial action in polymicrobial peritonitis (i.e. the striking enhancement of the innate immunity-mediated clearance of bacteria in the lungs, spleen, kidneys, and blood prior to antimicrobial therapy) by suppressing either Imp α5/SRTFs/ChREBPs or Imp β1/SREBPs transport. In Aim 2, we will analyze the mechanism of the survival gain through the timed and selective inhibition of Imp α5/SRTFs/ChREBPs or Imp β1/SREBPs transport without and with antimicrobial therapy. The new selective NTMs will significantly deepen our mechanistic understanding of innate immunity reprogramming using genomic and immunometabolic approaches. Thus, the new pathway-selective NTMs offer us a unique opportunity to (i) decode the transcriptional mechanism of septic shock that integrates microbial and metabolic inflammation and (ii) to develop clinically relevant strategies to reduce the incidence of septic shock, increase survival, and reduce long-term cognitive decline in survivors.

项目总结 核重编程对先天免疫和炎症的调节 我们建议推进我们对严重感染中先天免疫和炎症的机制研究 演变到最后阶段，感染性休克，从而威胁到战场上受伤士兵的生命，我们的 退伍军人和平民。65岁至85岁的退伍军人(和非退伍军人患者) 尤其容易发生感染性休克，死亡率高达45%，而许多幸存者的认知能力却很弱 拒绝。我们正在追求我们的长期目标，用一种新的类别来解码先天免疫和炎症 细胞穿透性核运输调节剂(NTM)，对炎症调节体进行重新编程。这其中的一个 抗炎肽对天然免疫介导的细菌具有前所未有的增强作用 肺(700倍)、脾(300倍)和血液(200倍)的清除，都发生在抗菌药之前 心理治疗。同时，它还改善了微血管内皮损伤的血液标志物，包括 血小板减少症。当与抗微生物治疗相结合时，NTM将存活率提高到55%， 相比之下，仅使用抗生素组的存活率为30%。这种名为cSN50.1的NTM多肽同时以 两艘核运输航天飞机，Importinsα5和β1。为了揭开先天免疫和 炎症是微生物和代谢紊乱的基础，我们设计、生产并测试了两种新的 单选择性NTMS：(I)导入α5-选择性cSN50.1α；(Ii)导入β-1-选择性cSN50.1β。令人惊讶的是， Importinα5-选择性多肽对内毒素休克小鼠的存活率为80% 内毒素是革兰氏阴性菌最强大的毒力因子之一，导致两种细菌 三分之一的感染性休克病例。这种新的NTM还有效地控制了西方饮食诱导的超敏反应 对致死性内毒素休克，同时防止肝糖原分解，这会导致高血糖和 与碳水化合物反应元件结合蛋白核转运相关的高甘油三酯血症 (CHREBPS)。反过来，Impβ1选择性多肽抑制了通过核运输介导的脂肪生成。 甾醇调节元件结合蛋白(SREBPs)控制着30多个参与生产的基因 甘油三酯、胆固醇和脂肪酸的含量，这是代谢综合征的标志。我们假设提议的 选择性NTMS与抗菌剂(S)联合使用可解决重症患者的多器官损害和死亡率 感染。此外，我们假设α5/SRTFS/CHREBPS和Impβ1/SREBPS的导入途径是 相互依赖，从而使患有代谢综合征的人更容易对严重的 感染。因此，在目标1中，我们将研究NTM在多菌种中的有益作用机制 腹膜炎(即通过先天免疫调节肺部细菌清除的显著增强， 脾、肾脏和血液)通过抑制Impα5/SRTFS/ChREBP或 IMPβ1/SREBPS传输。在目标2中，我们将从时间和时间两个方面分析生存收益的机制 抗菌药物对Impα5/SRTFS/ChREBPS或Impβ1/SREBPS转运的选择性抑制 心理治疗。新的选择性NTM将显著加深我们对先天免疫的机械性理解 使用基因组和免疫新陈代谢方法进行重新编程。因此，新的路径选择性非专利技术提供了 我们有一个独特的机会来(I)破译感染性休克的转录机制，它整合了微生物和 代谢性炎症和(Ii)制定临床相关策略，以减少感染性休克的发生率， 提高存活率，减少幸存者的长期认知衰退。