High-dimensional single cell profiling in immune-mediated glomerular diseases
Immune-mediated glomerular diseases are predominantly driven by pathogenic immune responses against renal autoantigens or by local manifestations of systemic autoimmunity. Kidney-infiltrating leukocytes are generally thought to play a critical role in this process, e.g. by local cytokine production and immune-epithelial cell interactions. However, our current knowledge of infiltrating leukocytes, as well as of their potential interaction with parenchymal kidney cells in patients with glomerulonephritis is very limited. Recent developments in single cell technologies have revolutionized the way we characterize complex immune cell assemblies, their dynamics, clonal distribution, pathways, function, and intercellular cross-talk. In this service project, we aim to provide high-dimensional single cell profiling to all CRC members. To this end, we established a workflow combining flow cytometry and single cell RNAsequencing (scRNA-seq) along with barcoded epitope measurement in glomerulonephritis patients “from sample preparation to data analysis”. This technical platform will provide all research groups of the CRC with the expertise to enable state-of-the-art single cell technologies, complemented with data analysis focused on the specific questions of each project. In particular, we will (1) offer high-dimensional single cell analysis of kidney biopsy specimen from GN patients; (2) provide integrated data analyses; (3) support the research projects of the CRC in single cell applications and bioinformatic analysis of experimental mouse models, and (4) further develop single cell technologies in our field including renal parenchymal cell isolation and analysis of cell-cell interaction studies. This service project will provide a comprehensive characterization of renal immune cells and epithelial cells in patients with selected forms of glomerulonephritis that will complement the Hamburg Glomerulonephritis Registry and be instrumental for the CRC 1192 to address specific questions in human glomerular disease.