Neutrophil Bioenergetic Metabolism in Sepsis

项目描述：本行政补充申请的目标是支持购买 的Agilent Technologies Seahorse XF Pro代谢分析仪（XF Pro），以确定中性粒细胞生物能量 这与题为“败血症的新方法”的母基金R35 GM 118337高度相关。 在脓毒症中维持器官功能”。脓毒症每年影响至少170万美国人， 27万名患者死亡，占所有医院死亡人数的30%。不幸的是 目前对于脓毒症患者没有特异性有效的治疗方法。我们发现细胞外冷诱导的 RNA结合蛋白（eCIRP）扩大了一种新的和破坏性的中性粒细胞群体，我们将其命名为抗原- 呈现老化中性粒细胞（APAN），从而显著增加脓毒症中的器官损伤和死亡。eCIRP具有 已显示引起线粒体生物发生和线粒体DNA片段化的变化。然而，在这方面， 尽管细胞能量代谢和线粒体生物合成在免疫中起着关键作用， 响应，eCIRP诱导的线粒体生物能量代谢变化对诱导的影响， APAN的激活和功能从未被研究过。XF Pro是一款高通量、顶级的 一种代谢分析仪，用于评估并允许操纵线粒体呼吸、糖酵解和ATP 活细胞的实时生产率。我们建议使用XF Pro进行线粒体研究， 通过询问三个与已经存在的问题有关的新问题，扩大母公司MIRA裁决的范围。 正在研究的父母R35奖：1）什么是代谢重编程的作用，在eCIRP诱导 APAN扩展？2)代谢重编程在APAN加重脓毒症中的作用是什么？3)并 靶向eCIRP通过抑制代谢重编程减轻败血症？XF Pro将 使实验能够显着扩大目前的理解，潜在的致病 机制，发现和评估新的潜在治疗靶点，并评估有前途的新治疗方法 对于败血症患者。

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.