Intraarticular microbial DNA as a novel mediator of osteoarthritis

关节内微生物 DNA 作为骨关节炎的新型介质

• 批准号: 10256679
• 负责人: Matlock Jeffries
• 金额: $ 42.39万
• 依托单位: OKLAHOMA MEDICAL RESEARCH FOUNDATION
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-08 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10256679
• 关键词: 16S ribosomal RNA sequencing; Acute; Adult; Affect; Age; Aging; Animal Model; Bacterial DNA; Biological Assay; Blood Cells; Cartilage; Cecum; Cells; Chondrocytes; Chronic; Clinical; Collaborations; DNA; DNA amplification; DNA delivery; Data; Degenerative polyarthritis; Development; Disease; Environment; Environmental Risk Factor; Epigenetic Process; Fostering; Functional disorder; Future; Germ-Free; Goals; Grant; Health; High Fat Diet; Histopathology; Human; Immune system; Immunophenotyping; Inflammation; Inflammatory; International; Intervention; Joints; Knee joint; Laboratories; Measures; Medial meniscus structure; Mediator of activation protein; Mission; Mus; Musculoskeletal Diseases; Obesity; Operative Surgical Procedures; Oral; Outcome; Pathogenesis; Patients; Pattern; Phase; Population; Positioning Attribute; Predisposition; Production; Public Health; Publishing; Reporting; Research; Research Personnel; Ribosomal RNA; Risk; Risk Factors; Role; Serum; Societies; Source; Testing; Therapeutic Agents; Time; Tissues; Transplantation; United States National Institutes of Health; Variant; Work; age related; aged; articular cartilage; bisulfite; chronic pain; cytokine; deep sequencing; diet-induced obesity; disability; epigenome; genome-wide; gut microbiome; human old age (65+); improved; innovation; insight; microbial; microbiome; microbiome research; microbiota; microbiota transplantation; mouse model; non-genetic; novel; peripheral blood; prevent; subchondral bone; success; synthetic polymer Bioplex; systemic inflammatory response; therapeutic development; transcriptome sequencing; transcriptomics; treatment strategy; whole genome

Project Summary / Abstract The objective of the proposed research is to better understand how cartilage microbial DNA patterns are established, and how these relate to osteoarthritis (OA) pathogenesis and progression. Previously published studies have outlined both gut microbiome pro-inflammatory changes with aging generally, but no work has yet been performed to examine how the gut and/or cartilage microbiomes, inflammation, and epigenetics intersect with OA. Our laboratory has previously examined in detail the epigenetic changes within cartilage and subchondral bone that are associated with OA development, and we have recently examined epigenetic changes within peripheral blood cells of human OA patients which are associated with increased risk of rapid OA progression. Our first Aim is to determine whether the cartilage microbial DNA pattern originates from the gut microbiome. To do this, we will populate the gut microbiome of germ-free mice with microbiota from young and OA-susceptible C57BL6/J (B6) mice then determine cartilage microbial DNA patterns at prespecified timepoints following inoculation. If successful, we will go on to test how variations in the cecal microbiome, associated with OA risk factors including aging and obesity, are reflected in changes of the cartilage microbiome profile. Furthermore, we will determine whether already-established cartilage microbiome patterns can be altered via changing the gut microbiome. Our second Aim will determine whether cartilage microbial DNA is sufficient to change the risk for OA development in mice. To do this, we will inject microbial DNA amplified from human OA patients and healthy controls into the knee joints of germ-free mice, then induce OA by disruption of the medial meniscus (DMM) surgery and measure histopathological outcome. If successful, we will then go on to determine local and systemic inflammatory cell population changes by CyTOF, serum cytokine changes by Bioplex assay, and epigenetic and transcriptomic changes on a genome-wide level following intraarticular microbial DNA delivery. The proposed work is important, as we do not have a full understanding of how non-genetic OA risk factors increases OA risk, nor do we understand how the microbiome influences OA risk. Our work is quite innovative in its use of germ-free mouse microbiome transplantation to evaluate OA and the use of cutting-edge CyTOF and epigenetic analyses. Finally, we will be the first to evaluate whether gut microbiome transplantation may be used as a therapeutic agent to prevent OA. Success in our proposal may open a new avenue for OA aging research and may offer a novel treatment strategy for OA.

项目摘要 /摘要 拟议研究的目的是更好地了解软骨微生物DNA模式如何 建立，以及这些与骨关节炎（OA）的发病机理和进展如何相关。先前发表 研究总体上概述了肠道微生物组的促炎性变化，但尚无工作 进行了检查肠道和/或软骨微生物，炎症和表观遗传学如何相交 与OA。我们的实验室先前已经详细研究了软骨内的表观遗传变化 与OA发育相关的软骨下骨，我们最近检查了表观遗传变化 在人OA患者的外围血细胞中，与快速OA的风险增加有关 进展。我们的第一个目的是确定软骨微生物DNA模式是否起源于 肠道微生物组。为此，我们将用年轻的微生物群填充无细菌小鼠的肠道微生物组 然后，在预先指定的情况下，确定软骨微生物DNA模式，并确定软骨微生物DNA模式 接种后的时间点。如果成功，我们将继续测试盲肠微生物组的变化， 与OA风险因素有关，包括衰老和肥胖，反映在软骨微生物组的变化中 轮廓。此外，我们将确定是否可以建立已经建立的软骨微生物组模式 通过更改肠道微生物组改变。我们的第二个目标将确定软骨微生物DNA是否是 足以改变小鼠OA发育的风险。为此，我们将从中注入微生物DNA 人OA患者和健康对照的无菌小鼠膝关节，然后通过中断诱导OA 内侧半月板（DMM）手术并测量组织病理学结果。如果成功，我们将继续 为了确定局部和全身性炎症细胞群体通过细胞变化，血清细胞因子通过 关节内的Bioplex分析以及整个基因组水平上的表观遗传和转录组变化 微生物DNA递送。拟议的工作很重要，因为我们对如何 非遗传OA风险因素会增加OA风险，我们也不了解微生物组如何影响OA风险。 我们的工作在使用无细菌小鼠微生物组移植以评估OA和 尖端细胞和表观遗传分析的使用。最后，我们将是第一个评估肠道的人 微生物组移植可用作预防OA的治疗剂。我们的提议中的成功可能会开放 OA衰老研究的新途径，可能为OA提供新颖的治疗策略。