Neutrophil Bioenergetic Metabolism in Sepsis

项目描述：此行政补充申请的目标是支持购买 安捷伦科技公司的赛马XF Pro代谢分析仪(XF Pro)用于测定中性粒细胞生物能量 脓毒症中的新陈代谢，这与家长基金R35GM118337题为“新的治疗方法”密切相关。 在脓毒症中维持器官功能“。败血症每年影响至少170万美国人，导致 27万名患者死亡，占所有医院死亡人数的30%。不幸的是，有一些 目前尚无针对败血症患者的特效治疗方法。我们发现细胞外的冷诱导 RNA结合蛋白(ECIRP)扩大了一种新的、具有破坏性的中性粒细胞群，我们将其命名为抗原- 出现老化的中性粒细胞(APAN)，从而显著增加脓毒症的器官损伤和死亡。ECIRP有 已被证明会导致线粒体生物发生和线粒体DNA片段化的变化。然而， 尽管细胞能量代谢和线粒体生物发生在免疫中起着关键作用 反应，eCIRP诱导的线粒体生物能量代谢的变化对诱导的影响， Apans的激活和功能从未被研究过。XF Pro是一款高吞吐量的顶级产品 代谢分析仪，评估并允许操纵线粒体呼吸、糖酵解和三磷酸腺苷 实时活细胞产生率。我们建议使用XF Pro进行的线粒体研究将显著 通过询问三个与已有的问题相关的新问题来扩大母公司Mira奖的范围 在Parent R35奖中正在研究：1)代谢重编程在eCIRP诱导中的作用是什么 APAN扩张？2)代谢重编程在APAN加重脓毒症中的作用是什么？3) 靶向eCIRP通过抑制代谢重编程减轻脓毒症？总而言之，XF Pro将 使实验能够显著扩展当前对潜在病因的理解 机制，发现和评估新的潜在治疗靶点，并评估有前景的新治疗方法 给脓毒症患者。

PROJECT DESCRIPTION: The goal of this Administrative Supplement application is to support the purchase of the Agilent Technologies Seahorse XF Pro metabolic analyzer (XF Pro) to determine neutrophil bioenergetic metabolism in sepsis, which is highly relevant to the parent grant R35GM118337 entitled "Novel Approaches to Maintaining Organ Function in Sepsis". Sepsis affects at least 1.7 million Americans annually, causing the death of 270,000 patients and being responsible for 30% of all hospital deaths. Unfortunately, there are currently no specific effective therapies for septic patients. We discovered that extracellular cold-inducible RNA-binding protein (eCIRP) expands a novel and damaging neutrophil population, which we named antigen- presenting aged neutrophils (APANs), thus markedly increasing organ injury and death in sepsis. eCIRP has been shown to cause changes in mitochondrial biogenesis and mitochondrial DNA fragmentation. However, even though cellular energy metabolism and mitochondrial biogenesis play a critical role in the immune response, the effects of eCIRP-induced changes in mitochondrial bioenergetic metabolism on the induction, activation, and function of APANs have never been studied. The XF Pro is a high-throughput, top-of-the-line metabolic analyzer that assesses and allows the manipulation of mitochondrial respiration, glycolysis, and ATP production rate of live cells in real-time. Our proposed mitochondrial studies with the XF Pro will significantly expand the scope of the parent MIRA award by interrogating three new questions related to those already being investigated in the parent R35 award: 1) What is the role of metabolic reprogramming in eCIRP-induced APAN expansion? 2) What is the role of metabolic reprogramming on APANs’ aggravation of sepsis? 3) Does targeting eCIRP mitigate sepsis via suppression of metabolic reprogramming? In summary, the XF Pro will enable experiments to significantly expand the current understanding of the underlying pathogenetic mechanisms, discover and evaluate novel potential therapeutic targets, and assess promising novel treatments for septic patients.