title: MiR-217 mediates the protective effects of the dopamine D2 receptor on fibrosis in human renal proximal tubule cells.
abstract: Lack or downregulation of the dopamine D2 receptor (D2R) increases the vulnerability to renal inflammation independent of blood pressure in mice. Common single nucleotide polymorphisms (SNPs) rs6276, 6277, and 1800497 in the human D2R gene are associated with decreased receptor expression/function and hypertension. Human renal proximal tubule cells from subjects carrying these SNPs have decreased D2R expression and increased expression of profibrotic factors and production of extracellular matrix proteins. We tested the hypothesis that the D2R mediates these effects by regulating micro-RNA expression. In cells carrying D2R SNPs, micro-RNAs (miRs)-217, miR-224, miR-335, and miR-1265 were downregulated, whereas miR-1290 was upregulated >4-fold compared with those carrying D2R wild-type alleles. However, only miR-217 was directly regulated by D2R expression. In cells carrying D2R wild-type, miR-217 inhibitor increased the expression of transforming growth factor (TGF)-β1, matrix metalloproteinase 3, fibronectin 1, and collagen 1a, whereas miR-217 mimic had the opposite effect. In cells carrying D2R SNPs, miR-217 mimic also decreased the expression of TGFβ1 and its targets. Wnt5a, a miR-217 target, was increased in cells carrying D2R SNPs and decreased by miR-217 mimic but increased by miR-217 inhibitor in both cell types. In cells carrying D2R wild-type, Wnt5a treatment increased TGFβ1 while silencing Ror2, a Wnt5a receptor, decreased TGFβ1 and blunted the Wnt5a-induced increase in cells carrying D2R wild-type. Our results show that renal proximal tubule cells from subjects carrying D2R SNPs resulting in D2R downregulation have increased TGFβ1 that is mediated by decreased regulation of the miR-217-Wnt5a-Ror2 pathway.
authors: From the Kidney Disease Center, First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, China (F.H., J.C.); Division of Nephrology, Department of Medicine (P.K., P.A.J., I.A.) and Department of Physiology (P.A.J.), University of Maryland School of Medicine, Baltimore; and Department of Pathology, University of Virginia School of Medicine, Charlottesville (J.G., R.A.F.) hanf8876@gmail.com.
authors: Fei
authors: F
authors: Han
authors: From the Kidney Disease Center, First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, China (F.H., J.C.); Division of Nephrology, Department of Medicine (P.K., P.A.J., I.A.) and Department of Physiology (P.A.J.), University of Maryland School of Medicine, Baltimore; and Department of Pathology, University of Virginia School of Medicine, Charlottesville (J.G., R.A.F.).
authors: Prasad
authors: P
authors: Konkalmatt
authors: From the Kidney Disease Center, First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, China (F.H., J.C.); Division of Nephrology, Department of Medicine (P.K., P.A.J., I.A.) and Department of Physiology (P.A.J.), University of Maryland School of Medicine, Baltimore; and Department of Pathology, University of Virginia School of Medicine, Charlottesville (J.G., R.A.F.).
authors: Jianghua
authors: J
authors: Chen
authors: From the Kidney Disease Center, First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, China (F.H., J.C.); Division of Nephrology, Department of Medicine (P.K., P.A.J., I.A.) and Department of Physiology (P.A.J.), University of Maryland School of Medicine, Baltimore; and Department of Pathology, University of Virginia School of Medicine, Charlottesville (J.G., R.A.F.).
authors: John
authors: J
authors: Gildea
authors: From the Kidney Disease Center, First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, China (F.H., J.C.); Division of Nephrology, Department of Medicine (P.K., P.A.J., I.A.) and Department of Physiology (P.A.J.), University of Maryland School of Medicine, Baltimore; and Department of Pathology, University of Virginia School of Medicine, Charlottesville (J.G., R.A.F.).
authors: Robin A
authors: RA
authors: Felder
authors: From the Kidney Disease Center, First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, China (F.H., J.C.); Division of Nephrology, Department of Medicine (P.K., P.A.J., I.A.) and Department of Physiology (P.A.J.), University of Maryland School of Medicine, Baltimore; and Department of Pathology, University of Virginia School of Medicine, Charlottesville (J.G., R.A.F.).
authors: Pedro A
authors: PA
authors: Jose
authors: From the Kidney Disease Center, First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, China (F.H., J.C.); Division of Nephrology, Department of Medicine (P.K., P.A.J., I.A.) and Department of Physiology (P.A.J.), University of Maryland School of Medicine, Baltimore; and Department of Pathology, University of Virginia School of Medicine, Charlottesville (J.G., R.A.F.).
authors: Ines
authors: I
authors: Armando
mesh_terms: D000483:Alleles; D002478:Cells, Cultured; D005355:Fibrosis; D005786:Gene Expression Regulation; D006801:Humans; D015151:Immunoblotting; D007674:Kidney Diseases / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D007687:Kidney Tubules, Proximal / Q000378:metabolism* / Q000473:pathology; D035683:MicroRNAs / Q000096:biosynthesis / Q000235:genetics*; D018613:Microscopy, Confocal; D012333:RNA, Messenger / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction; D017448:Receptors, Dopamine D2 / Q000235:genetics* / Q000378:metabolism; D015398:Signal Transduction
chemical_list: C553315:MIRN217 microRNA, human; D035683:MicroRNAs; D012333:RNA, Messenger; D017448:Receptors, Dopamine D2
keywords: microRNAs; receptor, dopamine D2; transforming growth factors