Cincinnati Center for Innovative Medicine in Adult & Pediatric Rheumatology

 描述(由申请人提供)：辛辛那提儿童医院医学中心和辛辛那提大学医学院建议在RFA-AR-14-016下建立一个网络综合临床和技术研究网站，由John Harley和Hermine Brunner领导。系统性红斑狼疮(SLE)将是我们网站的主要焦点，并将增加类风湿性关节炎(RA)和幼年特发性关节炎(JIA)的可自由扩展能力。我们的目标将是收集来自SLE和RA患者的材料和数据，并专注于SLE的肾脏和皮肤受累。我们将把单细胞RNA表达带到肾脏、皮肤，可能还有滑膜的分解细胞中。利用研究阶段0中已有的材料和数据，我们将制定方案和程序，分别评估狼疮性肾炎患者和对照组(肾母细胞瘤切除术后的正常肾)肾小球和间质细胞中单细胞信使RNA的表达；并了解这些操作和标准细胞冷冻方案的后果。我们将比较外周血中的细胞和狼疮性肾炎患者肾脏中的细胞，我们将分离成熟的B细胞和浆细胞进行单细胞RNA测序，以建立单个自身抗体的序列。我们计划在研究第一阶段进行三个试点实验：a.除了信使RNA外，还进行长非编码RNA的单细胞RNA测序；b.分解毛根细胞，并发展评估狼疮脱发中发现的单细胞RNA表达变化的能力；c.评估在尿液中发现的单个细胞的RNA表达模式，并将其与肾活检和外周血细胞中观察到的组织病理学进行比较。我们有初步数据显示，在狼疮性肾炎的强小鼠模型(NZB/W F1加补充IFNA)中，miR146a的混合制剂具有治疗效果，同时人类数据显示，miR146a的转基因表达减少了干扰素反应基因的表达。在研究第一阶段，我们将评估miR146a Agomir将肾小球细胞单个细胞RNA表达的扭曲恢复到非炎症状态的能力。最后，我们有一个强大的系统来提供临床样本，包括来自SLE和RA的肾脏、皮肤、头发和滑膜的组织活检，以及大量的医疗记录数据和来自其他研究的辅助数据，包括为本文建议的项目和我们网络同事的项目提供的基因分型。认识到由于该网络的审议，所进行的具体研究可能与所提议的有很大不同，我们提出了初步计划，为我们的网站在一个明显重要的领域提供科学重点。我们期待着参与网络领导委员会的审议，帮助网络的其他成员开展他们的项目，并在我们提出的项目中听取网络同事的建议、经验、帮助、参与和建议。

 DESCRIPTION (provided by applicant): The Cincinnati Children's Hospital Medical Center and the University of Cincinnati College of Medicine propose a Network Combined Clinical and Technology Research Site under RFA-AR-14-016 to be led by John Harley and Hermine Brunner. Systemic lupus erythematosus (SLE) will be the primary focus of our site with additional capabilities in rheumatoid arthritis (RA) and discretionary extension into juvenile idiopathic arthritis (JIA). Our goals will be to assemble materials and data from SLE and RA patients and focusing on kidney and skin involvement with SLE. We will bring single cell RNA expression to disaggregated cell of kidney, skin, and, potentially, synovium. With the materials and data available in Research Phase 0, we will develop protocols and procedures to evaluate separately the single cell messenger RNA expression in glomerular and interstitial renal cells from lupus nephritis cases and controls (normal kidney from Wilm's tumor nephrectomies); and to understand the consequences of these manipulations and of standard cell freezing protocols. We will compare cells in the peripheral blood to cells infiltrating the kidney with lupus nephritis We will isolate mature B and plasma cells for single cell RNA sequencing to establish the sequence of individual autoantibodies. We plan three pilot experiments in Research Phase 1: A. To perform single cell RNA sequencing with long non-coding RNA in addition to messenger RNA; B. To disaggregate cells from hair roots and develop the capacity to evaluate the changes in single cell RNA expression found in lupus alopecia; and C. To assess the RNA expression patterns of individual cells found in the urine and compare them to the histopathology observed on kidney biopsy and peripheral blood cells and disaggregated from the kidney biopsy. We have preliminary data showing therapeutic efficacy for an agomir of miR146a in a strong murine model for lupus nephritis (NZB/W F1 with supplemental IFNa) along with human data showing that the transfected expression of miR146a reduces expression of interferon responsive genes. In Research Phase 1 we will evaluate the capacity of miR146a Agomir to return the distortions in individual cell RNA expression of glomerular cells to a non-inflammatory state. Finally, we have a strong system for providing clinical samples, including tissue biopsies of kidney, skin, hair and synovium from SLE and RA, along with extensive medical record data and ancillary data from other studies, including genotyping, for the projects proposed herein and those of our Network colleagues. Realizing that the specific studies performed may, as a consequence of the deliberations of the Network, deviate substantially from what is proposed, we present tentative plans that provide our site with scientific focus in an area of obvious importance. We look forward to participating in the deliberations of the Network Leadership Committee, to assisting the other members of the network with their projects, and to the advice, experience, help, participation, and recommendations of our Network colleagues in the projects we propose.