Single cell analysis of healthy and diseased temporomandibular joint synovial fluid

项目摘要/摘要 颞下颌关节紊乱病(TMJDS)是一组复杂的、鲜为人知的疼痛状况， 深刻地影响着美国许多儿童和成年人的生活质量。出现TMJDS的患者经常 对保守治疗反应良好，但一些患者忍受着多次手术而没有缓解。 事实上，作为TMJDS基础的机械性知识的严重缺乏阻碍了 循证治疗。因此，迫切需要利用最新的技术进步 分子生物学，以开发新的治疗靶点，以改善护理。在这个项目中，我们建议 利用单细胞RNA测序(sc-RNAseq)对细胞群体和基因表达进行鉴定和分类 在TMJ滑液中。我们的中心假设是有症状的TMJD患者的滑液中含有一种 与TMJ疼痛直接相关的独特细胞群。在这个项目的UH2阶段，我们将开发 从健康和有症状的TMJD中获取高质量TMJ滑液和血液的方案 病人。在该项目的UH3阶段，我们将对TMJ进行单细胞RNA测序和分析 健康和有症状的TMJD患者的滑液和血液。我们假设滑液 有症状的TMJD患者含有一种独特的细胞群，可以通过单细胞RNA进行鉴定 在TMJD患者的血液中存在与疼痛相关但与关节紊乱无关的测序 紧接在症状恶化之前，在健康对照组中不存在。这项研究的结果 将在单细胞水平上提供有关TMJD的启动和解决的机械数据。

Project Summary/Abstract Temporomandibular joint disorders (TMJDs) are a set of complex, poorly understood painful conditions that profoundly impact the quality of life of many children and adults in the US. Patients presenting with TMJDs often respond well to conservative therapy, but some patients endure multiple surgical procedures without relief. Indeed, the profound lack of mechanistic knowledge underpinning TMJDs has hindered the development of evidence-based therapy. As a result, there is a critical need to utilize the most recent technological advances in molecular biology towards developing novel therapeutic targets to improve care. In this project, we propose to utilize single cell RNA sequencing (sc-RNAseq) to identify and classify the cell populations and gene expression in TMJ synovial fluid. Our central hypothesis is that synovial fluid from symptomatic TMJD patients contains a unique cell population that is directly correlated to TMJ pain. In the UH2 phase of this project, we will develop the protocols for obtaining high quality TMJ synovial fluid and blood from healthy and symptomatic TMJD patients. In the UH3 phase of this project, we will perform single cell RNA sequencing and analysis on TMJ synovial fluid and blood from both healthy and symptomatic TMJD patients. We hypothesize that synovial fluid from symptomatic TMJD patients contains a unique cell population that can be identified by single cell RNA sequencing, is correlated with pain but not joint derangement, is present in blood from TMJD patients immediately preceding worsening symptoms, and is not present in healthy controls. The results from this study will provide mechanistic data at the single cell level regarding the initiation and resolution of TMJD.