Immune Dysregulation in Sarcoidosis

摘要 结节病是一种特征不佳的免疫性疾病，通常被称为“伟大的模仿者” 由于它与其他多种疾病相似，诊断困难。疾病引起的 免疫细胞聚集物称为肉芽肿，在各种组织中形成，导致器官损伤和最终 严重者死亡。结节病的病因在很大程度上仍不清楚，没有明确的触发因素和症状 可以自行消失，或者尽管接受了治疗，情况仍会恶化。目前的研究揭示了与 一些常见的细菌种类以及免疫失调，特别是在T辅助1，T辅助17， 和T调节细胞群。然而，该领域的进展在一定程度上受到了复杂性的阻碍 目前模型中的疾病和局限性。 我们假设不同的T细胞亚群驱动免疫失调和进展 结节病。我们进一步假设，这种失调将结节病与其他肉芽肿区分开来。 处理并提供疾病特定建模和药物发现的基础。 AIM 1将应用最近开发的单细胞水平测序技术来更好地阐明 基因和细胞相互作用的复杂网络参与了发病机制。结节病患者的标本 肉芽肿和外周血单核细胞(PBMC)将通过单细胞RNA测序进行分析。 他们的转录本将与对照受试者的PBMC以及形成的对照肉芽肿进行比较 用纯化的蛋白衍生物刺激对照PBMC。细胞将通过它们的基因表达来识别 然后评估结节病患者与对照组相比在基因表达和调控方面的差异。 我们将利用这些数据建立结节病细胞类型和调控网络的转录图谱。 目的2将利用这一图谱评估成功利用患者的体外结节病模型 样本，以阐明疾病和对照之间的差异。使用每种模型创建的肉芽肿将是 通过单细胞rna-seq进行分析，并与我们的结节病转录图谱进行比较。这种方法促进了 对结节病免疫环境的详细评价和对现有能力的评价 模拟结节病的结节病模型。 该项目整合了测序和生物信息学的最新进展，以揭示机制 与结节病有关。结节病肉芽肿和外周血单核细胞的单细胞转录图谱 询问调节结节病免疫功能障碍的细胞相互作用，提供新的焦点 用于进一步的机械学研究。我们对结节病模型的分析将指导进一步的模型设计，以及 高通量结节病生物标志物和药物筛选的设计。这个项目有改进的潜力。 治疗的特异性和有效性，以及诊断的简便性。多学科的研究也 为我在探索一个关键的临床问题时成为一名内科科学家奠定了基础。

Abstract Sarcoidosis is a poorly characterized immune disorder that has often been called the “Great Imitator” due to its similarity to a variety of other diseases and the difficulty of diagnosis. The disease causes aggregates of immune cells called granulomas to form in various tissues, causing organ damage and eventual death in severe cases. Sarcoidosis etiology remains largely unknown, with no clear trigger and symptoms that can disappear on their own or worsen despite treatment. Current research has uncovered associations with some common bacterial species as well as immune dysregulation, especially in the T helper 1, T helper 17, and T regulatory cell populations. However, progress in the field has been hampered in part by the complexity of the disease and limitations in current models. We hypothesize that distinct subpopulations of T cells drive immune dysregulation and progression in sarcoidosis. We further hypothesize that this dysregulation differentiates sarcoidosis from other granulomatous processes and provides a basis for disease specific modeling and drug discovery. Aim 1 will apply recently developed single-cell level sequencing techniques to better elucidate the complex network of gene and cell interactions involved in pathogenesis. Samples of sarcoidosis patient granulomas and peripheral blood mononuclear cells (PBMCs) will be analyzed via single-cell RNA sequencing. Their transcriptomes will be compared to the PBMCs of control subjects as well as control granulomas formed by stimulation of control PBMCs with purified protein derivative. Cells will be identified by their gene expression profiles, then assessed for differences in gene expression and regulation in sarcoidosis compared to controls. We will use this data to develop a transcriptomic atlas of cell types and regulatory networks in sarcoidosis. Aim 2 will use this atlas to evaluate in vitro sarcoidosis models which successfully utilized patient samples to elucidate differences between disease and controls. Granulomas created using each model will be analyzed via single-cell RNA-seq and compared to our sarcoidosis transcriptomic atlas. This approach fosters detailed evaluation of the sarcoidosis immune environment and the evaluation of the capability of existing sarcoidosis models to mimic sarcoidosis. This project integrates recent advances in sequencing and bioinformatics to uncover mechanisms involved in sarcoidosis. The single-cell transcriptomic atlas of sarcoidosis granulomas and PBMCs will interrogate cellular interactions that regulate immune dysfunction in sarcoidosis, providing new points of focus for further mechanistic research. Our analysis of sarcoidosis models will guide further model design, and design of high throughput biomarker and drug screens for sarcoidosis. This project has the potential to improve the specificity and efficacy of treatment, as well as the ease of diagnosis. The multidisciplinary research also lays the groundwork for my training as a physician scientist while exploring a key clinical question.