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.
We will be good citizens of the new Accelerating Medicines Partnership (RFA-AR-14-016) and not only enhancing the work of our sister sites, but also bringing new technologies to the analysis of the pathology of lupus and rheumatoid arthritis and for the first time. In particular, we will evaluate the genes being turned on in every cell, all by itself, for example, of the filtering apparatus of the kidney, which will allow us to demonstrate which cells and which specific genes are causing these illnesses. The new technologies make possible new ways of testing and understanding new therapies, which will be demonstrated in this project.