Molecular and Cellular Dissection of Early Rheumatoid Arthritis

 描述（由申请人提供）：虽然在类风湿性关节炎（RA）的遗传定义和生物治疗方面取得了巨大进展，但仍然缺乏 关于这种疾病的发病机制和异质性的基本知识。最近的技术发展允许精确的分子分析疾病组织下降到单细胞水平。我们建议询问循环免疫细胞和滑膜组织细胞，以确定系统性免疫细胞多样性，滑膜病理和临床表型之间的分子和细胞相关性。在具体目标1（阶段0）中，我们将优化滑膜组织和外周血白细胞样本的制备和储存工艺，以支持各种先进的分析技术。这些包括滑膜组织细胞的单细胞RNA seq、表观遗传学研究（ATAC-Seq）、用于细胞表型的高维分析的CyTOF质谱细胞术以及至少12个外周血白细胞亚群的RNA seq分析。我们还将支持对滑膜组织组织切片进行原位分子和细胞分析的新技术的开发和可行性研究。此外，我们还将开发滑膜细胞系，以实现对滑膜细胞生物学的详细研究。在特定目标2（第1阶段）中，我们将招募30名类风湿关节炎受试者，使用在第0阶段开发和验证的技术进行血液和滑膜组织研究。对这些数据的系统级分析将推动在更大的患者队列中使用特定测定的选择。在特定目标3（2期）中，我们将对250例早期RA患者进行为期3年的观察性队列研究，分析滑膜组织和外周血白细胞。我们将采用0期和1期研究中出现的一组集中测定。将记录所有患者的病程、治疗和治疗反应。我们将解决以下假设：1）滑膜组织的多参数分析可以定义具有不同临床特征或结果的疾病子集; 2）使用高维分子和细胞数据分析外周血细胞子集，可以识别这些疾病子集的相关性; 3）可以开发可以识别以及预测治疗反应的有用参数。

 DESCRIPTION (provided by applicant): While tremendous progress has been made in genetic definition and biological treatments for rheumatoid arthritis (RA), there remains a lack of fundamental knowledge about the pathogenesis and heterogeneity of this disorder. Recent technological developments allow for precise molecular analysis of disease tissues down to the single cell level. We propose to interrogate circulating immune cells and synovial tissue cells to identify the molecular and cellular correlates between systemic immune cell diversity, synovial pathology and clinical phenotypes. In Specific aim 1 (phase 0), we will optimize processes for the preparation and storage of synovial tissue and peripheral blood leukocyte samples in order to support a variety of advanced analytic techniques. These include single cell RNA seq of synovial tissue cells, epigenetic studies (ATAC-Seq), CyTOF mass cytometry for high-dimensional analysis of cellular phenotypes, and RNA seq analysis of at least 12 peripheral blood leukoctye subsets. We will also support development and feasibility studies of novel technologies for in situ molecular and cellular analysis of histological sections of synovial tissu. In addition, we will develop synovial cell lines to enable detailed study of synovial cell biology.In Specific Aim 2 (phase 1), we will enroll 30 subjects with rheumatoid arthritis for study of blood and synovial tissue using the technologies developed and validated in phase 0. A systems level analysis of these data will drive the selection of specific assays to be utilized in a larger patiet cohort. In Specific Aim 3 (phase 2), we will carry out an observational cohort study of 250 early RA patients enrolled over a three year period with analysis of both synovial tissue and peripheral blood leukocytes. We will employ a set of focused assays that emerge from the studies in Phase 0 and 1. The disease course, therapy and response to therapy will be documented in all patients. We will address the following hypotheses: 1) multiparameter analysis of synovial tissue can define subsets of disease with distinct clinical features or outcomes; 2) analysis of peripheral blood cell subsets with high dimensional molecular and cellular data, can identify correlates for these disease subsets; 3) useful parameters can be developed that can identify, as well as predict, therapeutic response.