Cartilage Microbial Products as Novel Drivers of Knee Osteoarthritis Epigenetic Dysregulation

软骨微生物产品作为膝骨关节炎表观遗传失调的新驱动因素

• 批准号: 10588964
• 负责人: Matlock Jeffries
• 金额: --
• 依托单位: OKLAHOMA CITY VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-10-01 至 2026-09-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10588964
• 关键词: 16S ribosomal RNA sequencing; ATAC-seq; Adult; Affect; Age; Aging; Animal Model; Animals; Bacterial Genes; Cadaver; Cartilage; Cells; Chondrocytes; Chromatin; Chronic; Chronic Disease; Clinical; Clinical Data; Cytometry; DNA; DNA Methylation; DNA amplification; Data; Data Set; Degenerative polyarthritis; Development; Dietary Intervention; Disease; Epigenetic Process; Exposure to; Failure; Future; General Population; Germ-Free; Goals; Histologic; Human; In Vitro; Incidence; Inflammation; Inflammatory; Infrastructure; Joints; Knee; Knee Osteoarthritis; Laboratories; Link; Machine Learning; Measures; Medial meniscus structure; Mediating; Medical; Modeling; Mouse Strains; Mus; Musculoskeletal Diseases; Non obese; Obesity; Oligonucleotides; Operative Surgical Procedures; Outcome; Pain; Pathology; Pathway interactions; Patients; Pattern; Peripheral; Positioning Attribute; Predisposition; Publishing; Reporting; Research; Resistance; Risk; Risk Factors; Sampling; Serum; Severities; Shapes; Specific qualifier value; Specimen; Splenocyte; Synovial Membrane; Techniques; Testing; Therapeutic Agents; Tissues; Transplantation; Trauma; Veterans; Work; age related; behavioral outcome; bisulfite; bisulfite sequencing; chronic pain; cohort; cytokine; disability; draining lymph node; epigenome; fecal transplantation; genome-wide; gut microbiome; human old age (65+); immune activation; innovation; insight; knee replacement arthroplasty; machine learning model; microbial; microbial products; microbiome; microbiome alteration; microbiome analysis; microbiome research; microbiota transplantation; mouse model; next generation; next generation sequencing; non-genetic; normal aging; novel; novel therapeutic intervention; peripheral blood; prevent; recruit; subchondral bone; success; treatment strategy; whole genome

Project Summary / Abstract The objective of the proposed research is to better understand how age- and obesity-related gut microbiome changes are reflected in the knee cartilage microbiome and how these changes may be associated with knee osteoarthritis (OA) with a particular focus on epigenetics. OA is a leading cause of disability among Veterans and occurs at roughly 5x the rate of civilians. The incidence of OA rises with increases in age and obesity, and previously published studies have outlined gut microbiome changes with both age and obesity. Furthermore, OA in mice related to obesity can be lessened through dietary interventions that reshape the microbiome, and we have recently described a novel cartilage microbiome in humans and mice that changes with OA development. Our laboratory has previously examined in detail the epigenetic changes within cartilage, subchondral bone, and peripheral blood that are associated with OA development, and we have generated preliminary data that microbial DNA amplified from human OA cartilage can induce similar epigenetic changes in chondrocytes in vitro. In this project, our first Aim is to determine whether age- and obesity-related changes in the gut microbiome in human OA patients and healthy controls are reflected in similar changes in various joint microbiome niches, including cartilage, subchondral bone, and synovium. To do this, we will obtain paired cecal and cartilage samples from end- stage OA patients undergoing total knee replacement and matched control cadaveric samples from the NDRI. We will then profile microbiomes using 16s bacterial gene next-generation sequencing. We will then generate machine learning models of microbial changes associated with normal aging and obesity and compare these with OA-aging and OA-obesity. Our second Aim will determine whether age- and/or obesity-related cartilage microbiome changes impact OA outcomes and the cartilage microbiome specifically, using fecal microbiome transplantation (FMT) from human OA patients with and without aging and obesity into germ-free mice, and evaluating OA outcomes following DMM surgery. We will also examine both systemic and local inflammation associated with differences in microbiome transplants at prespecified timepoints using CyTOF. Our third Aim will evaluate epigenetic changes both within joint tissues and inflammatory cells induced by differences in the gut microbiome, using the same transplantation groups as in Aim 2. The proposed work is important, as we do not have a full understanding of why age and obesity are associated with increases in OA risk, nor do we understand how the cartilage microbiome influences OA risk. Our work is quite innovative in its use of paired gut and cartilage microbial samples, the next-generation techniques used to evaluate the microbiome, and our use of germ-free mouse microbiome transplantation to evaluate OA outcomes. We will also be the first to apply whole-genome bisulfite sequencing techniques to investigate the epigenetic changes within human and mouse cartilage that are altered by the local microbiome and associated with OA development. Finally, we will be the first to evaluate whether gut microbiome transplantation may be used as a therapeutic agent to alter age- and obesity-related OA risk. Success in our proposal may open a new avenue for OA aging research and may offer a novel treatment strategy for OA.

项目概要/摘要 拟议研究的目的是更好地了解年龄和肥胖相关的肠道如何 微生物组的变化反映在膝关节软骨微生物组中，以及这些变化是如何发生的 与膝骨关节炎（OA）相关，特别关注表观遗传学。 OA 是导致 退伍军人中的残疾率大约是平民的 5 倍。 OA 的发病率随 年龄和肥胖的增加，之前发表的研究概述了肠道微生物组的变化 年龄和肥胖。此外，与肥胖相关的小鼠的 OA 可以通过饮食来减少。 重塑微生物组的干预措施，我们最近描述了一种新型软骨微生物组 人类和小鼠随着 OA 的发展而变化。我们的实验室之前已经详细检查过 与 OA 相关的软骨、软骨下骨和外周血的表观遗传变化 开发，我们已经生成了从人类 OA 中扩增微生物 DNA 的初步数据 软骨可以在体外诱导软骨细胞发生类似的表观遗传变化。在这个项目中，我们的首要目标是 确定人类 OA 患者肠道微生物组的变化是否与年龄和肥胖相关 健康的控制反映在各种关节微生物生态位的类似变化，包括软骨、 软骨下骨和滑膜。为此，我们将从末端获取配对的盲肠和软骨样本 接受全膝关节置换术的阶段 OA 患者和来自该中心的匹配对照尸体样本 NDRI。然后，我们将使用 16s 细菌基因下一代测序来分析微生物组。我们将 然后生成与正常衰老和肥胖相关的微生物变化的机器学习模型， 将这些与 OA 衰老和 OA 肥胖进行比较。我们的第二个目标将确定年龄和/或 肥胖相关的软骨微生物组变化影响 OA 结果和软骨微生物组 具体来说，使用来自患有和不患有 OA 的人类 OA 患者的粪便微生物组移植 (FMT) 无菌小鼠的衰老和肥胖，并评估 DMM 手术后的 OA 结果。我们也会 检查与微生物组移植差异相关的全身和局部炎症 使用 CyTOF 预先指定的时间点。我们的第三个目标将评估关节内的表观遗传变化 使用相同的移植，由肠道微生物组差异诱导的组织和炎症细胞 目标 2 中的群体。拟议的工作很重要，因为我们还没有完全理解为什么年龄 和肥胖与 OA 风险增加相关，我们也不了解软骨微生物组如何 影响 OA 风险。我们的工作在使用配对的肠道和软骨微生物样本方面相当创新， 用于评估微生物组的下一代技术以及我们对无菌小鼠的使用 微生物组移植评估 OA 结局。我们也将率先应用全基因组 亚硫酸氢盐测序技术研究人类和小鼠软骨内的表观遗传变化 这些变化会受到当地微生物组的改变并与 OA 的发展相关。最后，我们将成为第一 评估肠道微生物组移植是否可以用作改变年龄和年龄的治疗剂 肥胖相关的 OA 风险。我们提案的成功可能会为 OA 老化研究开辟一条新途径，并可能 为 OA 提供了一种新的治疗策略。