A myogenic program is enacted by cardiac protein CSRP3 in human and mouse kidney

Introduction: Cardiac injury with renal injury (acute cardiorenal syndrome) can induce renal fibrosis, but mechanisms by which heart injury signals to kidney are poorly understood. The myocardiocyte differentiation factor cardiac LIM protein (CSRP3) is released into plasma after cardiac injury, and taken up by megalin (Lrp2) in renal proximal tubular epithelial cells in vivo. We evaluated the relationship between cardiac injury and renal fibrosis in a mouse model and in human cells by characterizing novel primary culture of human tubular epithelial cells and then testing the hypothesis that CSRP3 alters PTEC transcription of myogenesis, transport, and extracellular matrix genes in vitro and in vivo. Methods: In mice, cardiac arrest (CA) was induced with potassium chloride and resuscitated with epinephrine and chest compressions (CPR) as previously described. SnRNAseq was conducted on mouse kidney cells harvested 24h after CA/CPR and analyzed using Seurat. Research-designated human kidneys were obtained under IRB exemption for non-human subjects research and underwent dissection, digestion and FACS sorting for PTEC selection. hPTEC were passaged up to 4x before transporters were characterized by qPCR, immunoblot, and immunofluorescence. Recombinant human CSRP3 (Origene) was administered to hPTEC followed by RNASeq and qPCR. CSRP3 (5μg) was retroorbitally injected in mice and renal fibrosis was assessed 7 weeks later. Results: In mice, snRNAseq demonstrated that CA/CPR significantly upregulated muscle cell differentiation ontologies in proximal tubule cells, driven by regulation of 110/162 myogenesis genes. Following FACS sorting for CD10 and CD13, hPTEC were 95% pure PTEC and expressed RNA and protein for PT transporters Lrp2, Aqp1, Slc34a1, and Slc5a2. In hPTEC, CSRP3 induced myogenesis differentiation factor 1 (MyoD1)>30 fold, and 20 myogenesis pathway and extracellular matrix genes. 8/20 genes were coordinately regulated between mouse kidney after CA/CPR and hPTEC after CSRP3. CSRP3 injection caused tubulointerstitial fibrosis in mice (V αSMA /V kidney (%): 1.05 (CSRP3) vs 0.68 (control), n=8/group, p<0.01). Conclusion: hPTEC cultured from research-designated donor kidneys express a PT phenotype. The myocardiocyte differentiation factor CSRP3 causes transcriptional change in hPTEC, resulting in myogenic differentiation which is highly similar to that observed in mouse kidney after CA/CPR with coordinate regulation of myogenesis genes. Intravenous administration of CSRP3 to mice causes tubular interstitial fibrosis. Together these data strongly suggest CSRP3 may drive renal fibrosis via the myogenesis pathway. Merit grant I01BX004288-01 This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.