Single-cell omics approaches to investigate TMD

项目总结/摘要 颞下颌关节紊乱病（TMD）是最常见的口面疼痛形式，影响5-10%的成年人， 许多原因和危险因素都与TMD有关，但我们对生理因素的理解 慢性TMD疼痛发展的潜在机制是有限的。目前的理论框架 涉及由于炎症，免疫， 神经性和伤害性过程。进一步研究TMD中受影响的组织，包括肌肉， 咀嚼，颞下颌关节和滑膜，以及三叉神经分支，是必要的， 这些组织可以阐明病因学过程，但这些组织不易用于研究或诊断。 有证据支持来自血液和唾液的循环炎症和免疫介质的作用，但 这些潜在的生物标志物与口面组织病理学之间的关系需要严格地 通过直接比较的实验建立。我们建议同时评估 结果表明，TMD患者和对照组的咬肌组织和外周血中免疫细胞群的变化与 以便在细胞水平上确定病因学途径。在该项目的规划和可行性阶段，我们 将首先开发收集和处理组织的新技术，使最先进的单细胞 分析.在实施阶段，我们将使用血液中的单细胞RNA-seq评估基因转录， 将咬肌活检组织切片中的表达模式与空间转录组学进行比较。在咬肌 我们还将检查感觉神经密度和形态以研究神经病理机制， 和细胞表面蛋白来探索神经免疫相互作用。我们将识别出细胞模式， 病例，以及寻找代表不同亚类的个体之间的差异， 具有相似病因机制的患者。最后，我们将整合这些生物信息学数据集， 无偏见的组学方法，以发现具有预测和预后价值的生物标志物。该项目将 发展技术，提高对战区导弹防御系统的机理认识， 基于神经病学的分类，旨在更全面地了解整个 在细胞水平上的伤害感受性、伤害性和神经性机制的谱。

Project Summary/Abstract Temporomandibular disorders (TMD) are the most common form of orofacial pain, affecting 5-10% of adults in the U.S. Many causes and risk factors have been linked to TMD, but our understanding of the physiological mechanisms underlying the development of chronic TMD pain is limited. The current theoretical framework implicates sensitization at the peripheral and central levels due to a network of inflammatory, immune, neuropathic, and nociplastic processes. Further studies of the tissues affected in TMD, including muscles of mastication, the temporomandibular joint and synovium, and the trigeminal nerve branches, are necessary to elucidate etiological processes, but these tissues are not easily accessible for research or for diagnosis. Evidence supports a role for circulating inflammatory and immune mediators from blood and saliva, but the relationship between these potential biomarkers and pathology in orofacial tissues needs to be rigorously established in experiments with direct comparisons. We propose to conduct simultaneous assessments of masseter muscle tissue and immune cell populations in peripheral blood from both TMD cases and controls in order to identify etiological pathways at the cellular level. In the Planning and Feasibility phase of this project we will first develop novel techniques of collection and processing of tissues enabling state-of-the-art single-cell analysis. In the Implementation phase, we will assess gene transcription using single-cell RNA-seq in blood, comparing expression patterns with spatial transcriptomics in slices of masseter biopsy tissue. In masseter muscle we will also examine sensory nerve density and morphology to investigate neuropathic mechanisms, and cell-surface proteins to explore neuroimmune interactions. We will identify cellular patterns that distinguish cases from controls, as well as look for differences between individuals that represent distinct subcategories of patients with similar etiological mechanisms. Finally, we will integrate bioinformatics datasets from these unbiased omics approaches to discover biomarkers with predictive and prognostic value. This project will develop techniques and improve mechanistic understanding to advance the science of TMD from a rudimentary symptom-based classification toward a more complete understanding of biopsychosocial etiology across the spectrum of nociceptive, nociplastic, and neuropathic mechanisms at the cellular level.