Regulation of Innate Immunity and Inflammation Through Nuclear Reprogramming

项目摘要 通过核重编程调节天然免疫和炎症 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， 发展到他们的最终阶段，败血性休克，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。年龄在65至85岁之间的退伍军人（和非退伍军人患者） 特别容易发生败血性休克，死亡率为45%，而许多幸存者的认知能力下降， 下降我们正在追求我们的长期目标，用一类新的免疫调节剂来解码先天免疫和炎症。 细胞穿透核转运修饰剂（NTMs），重新编程炎症调节蛋白。其中一 抗炎肽实现了前所未有的先天免疫介导的细菌 在肺（700倍）、脾（300倍）和血液（200倍）中的清除率，均发生在抗菌剂使用前 疗法同时，它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，单纯化疗组的存活率为30%。该NTM肽cSN 50.1同时靶向 两个核转运穿梭蛋白，输入蛋白α5和β1。为了解开先天免疫的机制， 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型的 单选择性NTMs：（i）输入素α5-选择性cSN50.1α，和（ii）输入素β1-选择性cSN50.1β。引人注目的是， importin α5-选择性肽对内毒素休克的存活率为80%， 脂多糖（LPS）是革兰氏阴性菌的最强毒力因子之一，负责两种 三分之一的感染性休克病例这种新的NTM也有效地控制了西方饮食引起的过敏反应 致命的内毒素休克，同时防止肝糖原分解，这会导致高血糖症， 高甘油三酯血症与糖反应元件结合蛋白的核转运有关 （CHREBPs）。反过来，Imp β1-选择性肽抑制了由核转运介导的脂肪生成， 固醇调节元件结合蛋白（SREBP），控制30多个参与生产的基因， 甘油三酸酯、胆固醇和脂肪酸，代谢综合征的标志。我们假设， 使用选择性NTM与抗菌剂将解开多器官损伤和死亡率， 感染.此外，我们还证实Importin α5/SRTFs/CHREBPs和Imp β1/SREBPs通路是一个重要的信号通路。 相互依赖，从而使代谢综合征患者对严重的 感染.因此，在目标1中，我们将研究NTM在多微生物中的有益作用的机制。 腹膜炎（即先天免疫介导的肺部细菌清除的显著增强， 脾脏、肾脏和血液），通过抑制Imp α5/SRTFs/ChREBP或 Imp β1/SREBP转运。在目标2中，我们将通过定时和 在不使用和使用抗菌剂的情况下选择性抑制Imp α5/SRTFs/ChREBPs或Imp β1/SREBPs转运 疗法新的选择性NTMs将显著加深我们对先天免疫机制的理解 使用基因组和免疫代谢方法进行重编程。因此，新的路径选择性非关税措施提供了 我们有一个独特的机会，（i）解码感染性休克的转录机制，整合微生物和 代谢性炎症和（ii）开发临床相关策略以降低败血性休克的发生率， 提高生存率，减少幸存者长期认知能力下降。

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.