Modulating granulocytic myeloid-derived suppressor cell (G-MDSC) metabolic activity to promote Staphylococcus aureus biofilm clearance

调节粒细胞骨髓源性抑制细胞 (G-MDSC) 代谢活性以促进金黄色葡萄球菌生物膜清除

• 批准号: 10738662
• 负责人: Tammy L Kielian
• 金额: $ 23.03万
• 依托单位: UNIVERSITY OF NEBRASKA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-06-15 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10738662
• 关键词: Adoptive Transfer; Anti-Inflammatory Agents; Antibiotic Therapy; Antibiotics; Antiinflammatory Effect; Attenuated; Cardiac Myocytes; Cell physiology; Cells; Cellular Metabolic Process; Chronic; Coculture Techniques; Data; Development; Dyes; Endothelial Cells; Epithelial Cells; Excision; Exhibits; Exposure to; Extravasation; Future; Glycolysis; Glycolysis Inhibition; Goals; Health Care Costs; Human; Immune; Immune Evasion; Immunosuppression; Impaired cognition; Implant; In Vitro; Infection; Infiltration; Inflammatory; Inflammatory Response; Label; Laboratories; Leukocytes; Link; Macrophage; Mediating; Medical Device; Mesenchymal Stem Cells; Metabolic; Metabolic Pathway; Metabolism; Microbial Biofilms; Mitochondria; Modeling; Movement; Mus; Myeloid-derived suppressor cells; Myocardial Ischemia; Nanotubes; Operative Surgical Procedures; Oxidative Phosphorylation; PTPRC gene; Parkinson Disease; Pathologic; Pathology; Periprosthetic joint infection; Population; Property; Prosthesis; Reporting; Role; Shapes; Site; Source; Spinal cord injury; Staphylococcus aureus; T-Cell Activation; Tissues; Work; aerobic glycolysis; antibiotic tolerance; bactericide; cell type; chemotherapy; congenic; disability; granulocyte; humanized mouse; improved; in vivo; innovation; monocyte; mouse model; neutrophil; novel; novel strategies; novel therapeutic intervention; prevent; programs; superresolution microscopy; treatment strategy

Staphylococcus aureus (S. aureus) is a leading cause of biofilm-associated prosthetic joint infection (PJI) characterized by antibiotic tolerance and evasion of immune-mediated clearance. Our laboratory has established a critical role for granulocytic myeloid-derived suppressor cells (G-MDSCs), a pathologically activated neutrophil precursor, in attenuating monocyte/macrophage (MФ) proinflammatory properties and neutrophil bactericidal activity that leads to S. aureus biofilm persistence. The metabolic attributes of leukocytes are intimately linked with their inflammatory properties, relationships encompassing the emerging field of immunometabolism. This has been best described for MФs, where biases towards aerobic glycolysis or oxidative phosphorylation (OxPhos) dictate pro- vs. anti-inflammatory activity, respectively. In contrast, little information is available regarding the metabolic tendencies of G-MDSCs during infection and our preliminary studies are the first to demonstrate that G-MDSCs exhibit a glycolytic bias following S. aureus biofilm exposure in vitro and in vivo. Importantly, blocking glycolysis in G-MDSCs attenuated their suppressive activity resulting in decreased biofilm burden in vivo. This provides proof-of-concept that targeting G-MDSC metabolism is a tractable and novel approach to promote biofilm clearance. Recent studies have revealed that mitochondria can directly traffic between cells in vitro and in vivo via tunneling nanotubes (TNTs), where mitochondrial transfer in recipient cells promotes their OxPhos activity. However, most of these reports examined TNT-mediated mitochondrial transfer between mesenchymal stem cells and endothelial cells or cardiomyocytes, whereas this mechanism of intercellular metabolic rewiring has not been explored in the context of MФ-G-MDSC crosstalk. These studies will leverage MФs as a source of mitochondria for reprogramming G-MDSC metabolism, which originated from our prior work showing that MФ adoptive transfer reduced S. aureus biofilm burden in vivo. Indeed, our preliminary data support this innovative concept, where MФs transferred their mitochondria to G-MDSCs in a cell contact-dependent manner, which skewed G-MDSC metabolism towards OxPhos. This R21 revision will investigate the hypothesis that increasing mitochondrial abundance in G-MDSCs will result in metabolic reprogramming from glycolysis to an OxPhos bias coincident with diminished immune suppressive activity, resulting in improved biofilm clearance. Understanding how mitochondrial transfer from MФs can reprogram G- MDSC metabolism will be examined leveraging natural transfer and exogenous mitochondrial treatment paradigms in the following Specific Aims. 1) Identify the functional implications of mitochondrial transfer on G- MDSC anti-inflammatory activity and 2) Determine whether augmenting G-MDSC mitochondrial activity improves S. aureus clearance during PJI. These studies will inform our long-term goal of targeting critical metabolic nodes to reprogram aberrant leukocyte anti-inflammatory responses to eradicate biofilm together with antibiotics.

金黄色葡萄球菌（金黄色葡萄球菌）是生物膜相关关节感染（PJI）的主要原因 以抗生素耐受性和免疫介导的清除的进化为特征。我们的实验室已经建立 粒细胞髓细胞衍生的抑制细胞（G-MDSC）的关键作用，这是一种病理激活的中性粒细胞 前体，衰减单核细胞/巨噬细胞（MO）促炎特性和中性粒细胞杀菌性 导致金黄色葡萄球菌生物膜持久性的活性。白细胞的代谢属性密切相关 凭借其炎症特性，关系涵盖了免疫代谢的新兴领域。这 最好在MSS中描述，其中有氧糖酵解或氧化磷酸化偏见 （Oxphos）分别决定了抗炎活性。相反，几乎没有信息可用 关于在感染过程中G-MDSC和我们的初步研究的代谢趋势是第一个 证明G-MDSC在体外和体内暴露于金黄色葡萄球菌暴露后暴露了糖酵解偏置。 重要的是，阻塞G-MDSC中的糖酵解减弱了其抑制活性，导致生物膜降低 体内负担。这提供了概念证明，靶向G-MDSC代谢是一种可挑剔且新颖的 促进生物膜清除的方法。最近的研究表明，线粒体可以直接流动 通过隧道纳米管（TNT）在体外和体内细胞之间，其中线粒体转移在受体细胞中 促进其OXPHOS活性。但是，这些报告中的大多数检查了TNT介导的线粒体转移 间充质干细胞和内皮细胞或心肌细胞之间 在MO-G-MDSC串扰的背景下，尚未探索细胞间代谢重新布线。这些研究 将利用MOS作为线粒体的来源来重编程G-MDSC代谢，起源于 我们先前的工作表明，MOS适应性转移减少了体内的金黄色葡萄球菌生物膜伯嫩。确实，我们的 初步数据支持这一创新概念，其中MOS将其线粒体转移到了G-MDSC中 细胞接触依赖性方式，将G-MDSC代谢偏向OXPHOS。此R21修订将 研究以下假设：G-MDSC中的线粒体丰度增加将导致代谢 从糖酵解到与免疫抑制活性减少的OXPHOS偏置重新编程， 导致生物膜清除率提高。了解如何从MOS进行线粒体转移可以重新编程G- 将检查MDSC代谢，以利用天然转移和外源线粒体治疗 以下特定目标中的范例。 1）确定线粒体转移对G-的功能意义 MDSC抗炎活性和2）确定增强G-MDSC线粒体活性是否会改善 S. PJI期间的金黄色葡萄球菌清除。这些研究将为我们的长期目标提供针对关键代谢节点的长期目标 为了重新编程对放射性生物膜的异常白细胞抗炎反应以及抗生素。