Molecular investigation of the effects of the combination of cyclophosphamide used in the treatment of glomerulonephritis and the TRPC1/6 inhibitor DGsMTx4 on podocyte survival.

Project no. 22448 · Status: Kapanmış (Yayın bekleniyor) · 2021

Summary

Glomerulonephritis is an inflammatory disease caused by a series of immune-mediated disorders that lead to inflammation in the glomerulus and other parts of the kidney. Cyclophosphamide, in addition to its immunosuppressive and immunomodulatory effects, also possesses antimitotic and antireplicative properties, making it a standard treatment applied in low doses for glomerulonephritis. Podocytes are unique cells with specialized foot processes connected by slit diaphragms located on the outer side of glomerular capillaries. Proteinuria is an early consequence of podocyte damage and is a typical sign of kidney disease. Persistent podocyte damage eventually leads to chronic kidney disease. Calcium (Ca2+) signaling plays a critical role in podocyte damage. Transient receptor potential cation (TRPC) channels have been shown to contribute to calcium entry in podocytes. To investigate the molecular effects of the commonly used immunosuppressant cyclophosphamide and the TRPC1/6 inhibitor D-GsMTx4 agent in treatment, we conducted research on human kidney and urine-derived podocyte cell lines and human proximal tubular epithelial cell lines in vitro. Our results showed significant decreases in the following genes in PODO-152 ADR-damaged cells following combinations of Cyclophosphamide and D-GsMTx4: ABL1, CRADD, DIABLO, TRAF2, XIAP, BAK1, BCL2A1, NOD1, RIPK2, BID, APAF1, NFKB1, TP53, BCL2L11, CASP4, IGF1R, BAG1, BRAF, DAPK1, TRAF3, BFAR, TP73, CASP7, CYCS, TP53, BP2, CD27, AKT1, TNF, TNFRSF10A, and TNFRSF11B; and significant upregulation of apoptotic-related FAS, FASLG, NAIP, CASP5, and AIFM1 genes was observed. In PODO-256 ADR-damaged cells, the combinations of Cyclophosphamide and D-GsMTx4 resulted in significant decreases in the LTA, ABL1, NAIP, TP53, AIFM1, DIABLO, TNF, CYCS, BID, TRAF3, TP73, and BAK1 genes, and significant upregulation of BCL2A1, IGF1R, and CASP8 genes was determined. In ADR-damaged PODO-152 and PODO-256 cell lines, after 48 hours of treatment with Cyclophosphamide + D-GsMTx4 combination doses, apoptotic cells were observed at rates of 1.7% and 2.0%, respectively.

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