An integrative study of circulating leukocyte composition, epigenetic patterns, and functional consequences in knee osteoarthritis

膝骨关节炎循环白细胞组成、表观遗传模式和功能后果的综合研究

• 批准号: 10267470
• 负责人: Matlock Jeffries
• 金额: $ 10.01万
• 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: 2020-08-01 至 2022-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10267470
• 关键词: integrative; study; circulating; leukocyte; composition

Project Summary / Abstract My previous research in epigenetics and clinical training as a rheumatology fellow have prepared me well for a future career as a physician scientist. The K08 proposal described herein will allow me the necessary protected time and mentoring to expand my skills and achieve my primary goals of becoming leader in the field of osteoarthritis (OA) epigenetics. I will participate in formal coursework with a particular focus on bioinformatics, statistics, and research communication at the University of Oklahoma Health Sciences Center. I will expand my presentation skills through both didactic training and presentation of my research findings. My career development will be greatly advanced by my appointment as an Oklahoma Shared Clinical and Translational Resources Scholar, as well as regular meetings with my primary mentor and advisory committee. My research during this pivotal period will focus on OA, a chronic, debilitating musculoskeletal disease affecting 40% of US adults over the age of 70. Understanding the mechanisms of gene-environmental interaction and the contributions of extraarticular tissues to OA are key to the development of clinical biomarkers and therapeutic agents. I have published data showing significantly altered DNA methylation patterns in OA cartilage and subchondral bone, and have preliminary data from peripheral blood mononuclear cells of OA patients demonstrating remarkably consistent epigenetic dysregulation of several immune-related pathways. Based on these data, I hypothesize that OA is a disease characterized by low-level systemic inflammation, associated with dysregulated epigenomes that alter gene transcription, leading to disease pathogenesis. To evaluate this, I will recruit knee OA patients at risk of radiographic progression and perform serial blood draws and knee X-rays for later analysis. I will determine if changes in circulating inflammatory cell populations and/or their response to stimulation are present in knee OA patients compared to both healthy and rheumatoid arthritis controls (Aim 1), and examine whether patterns in inflammatory cell composition are predictive of radiographic OA progression over time, using state-of-the-art cyTOF analysis. I will then confirm an altered DNA methylation pattern using banked DNA from a large OA cohort (Aim 2a). Subsequently, I will study DNA methylation and gene transcription in leukocyte subsets: CD4+, CD8+, and monocyte cells (Aim 2b). Finally, I will use a novel, targetable dCas9-TET1 fusion protein to evaluate the functional consequences of specific DNA demethylation events I previously identified in my OA cartilage work (Aim 3). Discoveries achieved by these aims will form the preliminary data for strong R01 applications to confirm novel diagnostic biomarkers and develop new therapeutic strategies to treat this devastating disease. If successful, I will provide the first data regarding circulating OA inflammatory cell subpopulations. Leveraging the core facilities at OMRF, I will additionally provide a publically-available biorepository of the specimens and data I collect for future study.

项目概要/摘要 我之前在表观遗传学方面的研究和作为风湿病学研究员的临床培训为我做好了充分的准备 未来的职业是一名医师科学家。本文描述的 K08 提案将允许我获得必要的保护 时间和指导来扩展我的技能并实现成为该领域领导者的主要目标 骨关节炎（OA）表观遗传学。我将参加正式课程，特别关注生物信息学， 俄克拉荷马大学健康科学中心的统计和研究交流。我会扩展我的 通过教学培训和研究成果的展示来提高演讲技巧。我的职业生涯 我被任命为俄克拉荷马州共享临床和转化人员，这将极大地促进发展 资源学者，以及与我的主要导师和咨询委员会的定期会议。 在这个关键时期，我的研究将集中在 OA，一种慢性、衰弱性肌肉骨骼疾病，影响 40% 的美国成年人年龄超过 70 岁。了解基因与环境相互作用的机制以及 关节外组织对 OA 的贡献是临床生物标志物和治疗方法开发的关键 代理。我发表的数据显示 OA 软骨中 DNA 甲基化模式发生显着改变 软骨下骨，并有 OA 患者外周血单个核细胞的初步数据 证明了几种免疫相关途径的非常一致的表观遗传失调。基于 根据这些数据，我假设 OA 是一种以低水平全身炎症为特征的疾病，与 表观基因组失调会改变基因转录，导致疾病发病机制。 为了评估这一点，我将招募有放射学进展风险的膝关节 OA 患者并进行系列血液检查 绘图和膝盖X光片以供以后分析。我将确定循环炎症细胞群是否发生变化 和/或与健康人和类风湿患者相比，膝关节骨关节炎患者对刺激的反应也存在 关节炎对照（目标 1），并检查炎症细胞组成的模式是否可以预测 使用最先进的 cyTOF 分析来观察放射学 OA 随着时间的推移的进展情况。然后我将确认更改 使用来自大型 OA 队列的库 DNA 进行 DNA 甲基化模式（目标 2a）。接下来我会研究DNA 白细胞亚群中的甲基化和基因转录：CD4+、CD8+ 和单核细胞（目标 2b）。最后，我 将使用一种新型的、可靶向的 dCas9-TET1 融合蛋白来评估特定 DNA 的功能后果 我之前在 OA 软骨工作中发现的去甲基化事件（目标 3）。 通过这些目标实现的发现将形成强大的 R01 应用的初步数据，以确认新颖性 诊断生物标志物并开发新的治疗策略来治疗这种毁灭性的疾病。如果成功的话我 将提供有关循环 OA 炎症细胞亚群的第一个数据。发挥核心作用 OMRF 设施，我将另外提供一个公开可用的样本和数据生物存储库 收集以供将来学习。