Innate Immune Response to S. aureus Biofilm

对金黄色葡萄球菌生物膜的先天免疫反应

基本信息

批准号：
10665032
负责人：
Tammy L Kielian
金额：
$ 58.82万
依托单位：
UNIVERSITY OF NEBRASKA MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-07-01 至 2024-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10665032
关键词：
Acetylation Anabolism Anti-Bacterial Agents Anti-Inflammatory Agents Antibiotics Cells Cellular Metabolic Process ChIP-seq Data Development Disease Environment Excision Funding Genus staphylococcus Glycolysis Goals HDAC4 gene Hip region structure Histone Deacetylase Inhibitor Histones Immune Immune response Impairment Implant Incidence Infection Infiltration Inflammatory Innate Immune Response Interleukin-10 Knockout Mice Laboratories Leukocytes Libraries Macrophage Metabolic Metabolic Pathway Metabolism Microbial Biofilms Modeling Molecular Molecular Mechanisms of Action Mus Myeloid-derived suppressor cells Nebraska Neutrophil Infiltration Oligomycins Oxidative Phosphorylation Pathway interactions Patients Periprosthetic joint infection Phenotype Production Property Prosthesis Role Source Staphylococcus aureus Translating Urokinase Work aerobic glycolysis genome-wide improved in vivo inhibitor innovation knee replacement arthroplasty metabolome metabolomics monocyte mutant nanoparticle neutrophil programs promoter recruit

项目摘要

Staphylococcus aureus (S. aureus) is a leading cause of biofilm-associated prosthetic joint infections (PJIs) typified by an anti-inflammatory cellular milieu. The inflammatory phenotype of leukocytes is intimately tied to their metabolic status, where anti-inflammatory MΦs primarily rely on oxidative phosphorylation (OxPhos) and pro-inflammatory MΦs utilize glycolysis. Monocytes are also polarized toward an anti-inflammatory state during S. aureus PJI, which our preliminary data shows coincides with an OxPhos bias. We have devised an innovative approach to metabolically re-reprogram biofilm-associated monocytes to promote glycolysis using cell-targeted nanoparticles containing the OxPhos inhibitor oligomycin. Treatment of established biofilms with oligomycin nanoparticles promoted monocyte pro-inflammatory activity concomitant with increased neutrophil recruitment, leading to biofilm clearance. During the recent PPG cycle, our laboratory was the first to identify that myeloid- derived suppressor cells (MDSCs) skew biofilm-associated monocytes toward an anti-inflammatory state, in part, through IL-10 production. Therefore, we screened the Nebraska Transposon Mutant Library to identify mutants impaired in their ability to trigger IL-10 production. Significant hits involved in lactate biosynthesis were identified, and our preliminary data support a role for S. aureus-derived lactate in organizing the anti-inflammatory biofilm milieu, progressing from MDSCs/monocytes as a target to defining the molecular mechanism of action. First, during PJI, D- and L-lactate levels are reduced in S. aureus ddh1 and ldh1/ldh2 mutants, respectively, concomitant with significant reductions in MDSC infiltrates and IL-10 production, which translates into enhanced leukocyte recruitment, and biofilm clearance. Second, the IL-10 promoter is activated by acetylation and our ChIP-Seq data demonstrate that histone promoter acetylation is dramatically increased genome-wide in leukocytes recovered from WT vs. ldh1/ldh2 infected mice, providing molecular evidence that S. aureus biofilm- derived lactate functions as a histone deacetylase inhibitor (HDACi). Our central P01 hypothesis is that S. aureus biofilm development creates unique metabolic niches that promote an immune suppressive environment. In Project 4, we will explore the existence of a metabolic triad between S. aureus biofilm, MDSCs, and monocytes, whereby biofilm-derived lactate promotes leukocyte anti-inflammatory properties, in part, via IL- 10 production, contributing to biofilm persistence. This metabolic crosstalk and the molecular mechanisms responsible for this interplay will be explored in the following Specific Aims. 1) Establish that leukocyte metabolism can be targeted in vivo to promote pro-inflammatory activity and biofilm clearance; 2) Investigate the role of S. aureus biofilm-derived lactate in promoting MDSC and monocyte anti-inflammatory activity by stimulating IL-10 production; and 3) Determine whether S. aureus biofilm-derived lactate regulates IL-10 production by inhibiting histone deacetylase (HDAC) activity. These studies will inform our long-term goal of targeting metabolic pathways that disarm anti-bacterial innate immune defenses to facilitate biofilm eradication.

金黄色葡萄球菌 (S. aureus) 是生物膜相关假体关节感染 (PJI) 的主要原因以抗炎细胞环境为代表。白细胞的炎症表型与此密切相关。他们的代谢状态，其中抗炎 MΦ 主要依赖于氧化磷酸化 (OxPhos) 和促炎 MΦ 在糖酵解过程中也会极化为抗炎状态。我们的初步数据显示金黄色葡萄球菌 PJI 与 OxPhos 偏差一致，我们设计了一种创新方法。使用细胞靶向代谢重新编程生物膜相关单核细胞以促进糖酵解的方法含有 OxPhos 抑制剂寡霉素的纳米颗粒用寡霉素处理已建立的生物膜。纳米粒子促进单核细胞促炎活性，同时增加中性粒细胞募集，导致生物膜清除在最近的 PPG 周期中，我们的实验室是第一个发现骨髓- 衍生抑制细胞（MDSC）部分地将生物膜相关单核细胞偏向抗炎状态，因此，我们筛选了内布拉斯加州转座子突变体库来鉴定突变体。确定了其触发 IL-10 产生的能力受损，我们的初步数据支持金黄色葡萄球菌衍生的乳酸在组织抗炎生物膜中的作用首先，从 MDSC/单核细胞作为目标到定义分子作用机制。在 PJI 期间，金黄色葡萄球菌 ddh1 和 ldh1/ldh2 突变体中的 D- 和 L- 乳酸水平分别降低，伴随着 MDSC 渗透和 IL-10 产生的显着减少，这转化为增强白细胞募集和生物膜清除其次，IL-10 启动子通过乙酰化和我们的作用而被激活。 ChIP-Seq 数据表明组蛋白启动子乙酰化在全基因组范围内显着增加从 WT 与 ldh1/ldh2 感染小鼠中回收的白细胞，提供了金黄色葡萄球菌生物膜-的分子证据衍生的乳酸作为组蛋白脱乙酰酶抑制剂 (HDACi) 发挥作用。我们的中心 P01 假设是 S. 金黄色葡萄球菌生物膜的发育创造了独特的代谢生态位，促进免疫抑制在项目 4 中，我们将探索金黄色葡萄球菌生物膜、MDSC、和单核细胞，因此生物膜衍生的乳酸部分通过 IL- 促进白细胞抗炎特性 10 的产生，有助于生物膜的持久性和分子机制。负责这种相互作用的因素将在以下具体目标中进行探讨： 1) 确定白细胞。可以靶向体内代谢以促进促炎活性和生物膜清除；2) 研究金黄色葡萄球菌生物膜衍生的乳酸在促进 MDSC 和单核细胞抗炎活性中的作用刺激 IL-10 产生；和 3) 确定金黄色葡萄球菌生物膜衍生的乳酸是否调节 IL-10 这些研究将为我们的长期目标提供信息。针对解除抗菌先天免疫防御的代谢途径，以促进生物膜的根除。