Regulation of glomerular matrix proteins via podocytic and endothelial-cell derived microRNAs

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Proteinuria is a hallmark of glomerular disease. We identified nephronectin (NPNT) as an extracellular matrix protein important for the glomerular filtration barrier. Podocyte microRNA-378a-3p (miR-378a-3p) and endothelial miR-192 regulate podocyte NPNT expression and are upregulated in membranous glomerulonephritis. Overexpression of miR-378a-3p or miR-192 as well as knockdown of npnt caused a phenotype with proteinuria, edema, podocyte effacement and widening of the glomerular basement membrane. Furthermore, we want to analyze the regulation of glomerular matrix proteins via podocyte and endothelial cell derived miRs.We will investigate the glomerular function of npnt and generate podocyte specific npnt knockout mice that will be analyzed regarding spontaneous or stress related development of glomerular damage. Using different Npnt constructs with and without the specific miR binding site we will solve the question to what degree the glomerular changes after miR-378a-3p overexpression are due to npnt knockdown or other miR targets.We will use fluorescent dextrans with different molecular weights to characterize the increase of glomerular permeability after npnt knockdown. To identify other matrix associated targets that are post-transcriptionally controlled by miR-378a-3p and miR-192, we will perform comparative microarrays after miR transfection of human podocytes and glomerular endothelial cells. Finally, we will investigate the therapeutic potential of antagomirs of miR-378a-3p and miR-192 to analyze if inhibition of these miRs has a protective effect.