Neutrophil Bioenergetic Metabolism in Sepsis

点击翻译按钮获取中文摘要

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.