title: Renal endothelial protein C receptor expression and shedding during diabetic nephropathy.
abstract: Essentials Endothelial protein C receptor (EPCR) promotes diabetic nephropathy (DN) outcome improvement. Renal expression and shedding of EPCR were measured in diabetic patients with or without DN. Inhibition of metalloproteinase-driven EPCR shedding restored glomerular endothelium phenotype. EPCR shedding through metalloproteinase ADAM17 contributes to the worsening of DN.
  
  
  
  Background Diabetic nephropathy (DN) represents the leading cause of end-stage renal disease. The endothelial protein C receptor (EPCR) and its ligand (activated protein C) have been shown to ameliorate the phenotype of DN in mice. EPCR activity can be regulated by proteolytic cleavage involving ADAMs, yielding a soluble form of EPCR (sEPCR). Objective To characterize the renal expression and shedding of EPCR during DN. Methods EPCR levels were measured in plasma, urine and biopsy samples of diabetic patients with (n = 73) or without (n = 63) DN. ADAM-induced cleavage of EPCR was investigated in vitro with a human glomerular endothelium cell line. Results DN patients showed higher plasma and urinary levels of sEPCR than diabetic controls (112.2 versus 135.2 ng mL(-1) and 94.35 versus 140.6 ng mL(-1) , respectively). Accordingly, glomerular endothelial EPCR expression was markedly reduced in patients with DN, and this was associated with increased glomerular expression of ADAM-17 and ADAM-10. In vitro, EPCR shedding was induced by incubation of glomerular endothelium in high-glucose medium, and this shedding was suppressed by ADAM-17 inhibition or silencing, which led to improved vascular endothelial cadherin (VE-cadherin) expression and reduced mRNA expression of transforming growth factor (TGF)-β. In addition, EPCR silencing led to minor effects on VE-cadherin but to a significant increase in TGF-β mRNA expression. Conclusion Inhibition of ADAM-driven glomerular EPCR shedding restored the endothelial phenotype of glomerular endothelium, whereas EPCR silencing led to enhanced expression of TGF-β, a marker of endothelial-mesenchymal transition. These findings demonstrate that EPCR shedding driven by ADAMs contributes to the worsening of DN.
authors.affiliation: Department of Pathology, Academic Medical Center, University of Amsterdam, Amsterdam, the Netherlands.
authors.forename: L
authors.initials: L
authors.lastname: Lattenist
authors.affiliation: Department of Pathology, Academic Medical Center, University of Amsterdam, Amsterdam, the Netherlands.
authors.forename: P
authors.initials: P
authors.lastname: Ochodnický
authors.affiliation: Department of Internal Medicine, Slotervaart Hospital, Amsterdam, the Netherlands.
authors.forename: M
authors.initials: M
authors.lastname: Ahdi
authors.affiliation: Department of Pathology, Academic Medical Center, University of Amsterdam, Amsterdam, the Netherlands.
authors.forename: N
authors.initials: N
authors.lastname: Claessen
authors.affiliation: Department of Pathology, Academic Medical Center, University of Amsterdam, Amsterdam, the Netherlands.
authors.forename: J C
authors.initials: JC
authors.lastname: Leemans
authors.affiliation: Academic Renal Unit, University of Bristol, Bristol, UK.
authors.forename: S C
authors.initials: SC
authors.lastname: Satchell
authors.affiliation: Department of Pathology, Academic Medical Center, University of Amsterdam, Amsterdam, the Netherlands.|Department of Pathology, Radboud University Nijmegen Medical Center, Nijmegen, the Netherlands.
authors.forename: S
authors.initials: S
authors.lastname: Florquin
authors.affiliation: Department of Internal Medicine, Slotervaart Hospital, Amsterdam, the Netherlands.|Department of Experimental Vascular Medicine, Academic Medical Center, University of Amsterdam, Amsterdam, the Netherlands.
authors.forename: V E
authors.initials: VE
authors.lastname: Gerdes
authors.affiliation: Department of Pathology, Academic Medical Center, University of Amsterdam, Amsterdam, the Netherlands.
authors.forename: J J T H
authors.initials: JJ
authors.lastname: Roelofs
mesh_terms: D000072197:ADAM10 Protein / Q000378:metabolism; D000072198:ADAM17 Protein / Q000378:metabolism; D000368:Aged; D053829:Amyloid Precursor Protein Secretases / Q000378:metabolism; D001706:Biopsy; D002460:Cell Line; D003920:Diabetes Mellitus / Q000097:blood / Q000473:pathology / Q000652:urine; D003924:Diabetes Mellitus, Type 2 / Q000378:metabolism / Q000473:pathology; D003928:Diabetic Nephropathies / Q000378:metabolism* / Q000473:pathology; D000074283:Endothelial Protein C Receptor / Q000378:metabolism*; D004727:Endothelium / Q000473:pathology; D005260:Female; D020868:Gene Silencing; D006801:Humans; D007668:Kidney / Q000378:metabolism*; D007678:Kidney Glomerulus / Q000378:metabolism; D008024:Ligands; D008297:Male; D008565:Membrane Proteins / Q000378:metabolism; D045726:Metalloproteases / Q000378:metabolism; D008875:Middle Aged; D010641:Phenotype; D034741:RNA, Small Interfering / Q000378:metabolism; D016212:Transforming Growth Factor beta / Q000378:metabolism
chemical_list: D000074283:Endothelial Protein C Receptor; D008024:Ligands; D008565:Membrane Proteins; C478370:PROCR protein, human; D034741:RNA, Small Interfering; D016212:Transforming Growth Factor beta; D053829:Amyloid Precursor Protein Secretases; D045726:Metalloproteases; D000072197:ADAM10 Protein; C491236:ADAM10 protein, human; D000072198:ADAM17 Protein; C000606462:ADAM17 protein, human
keywords: ADAM-17; albuminuria; diabetes complications; diabetic nephropathy; endothelial cell protein C receptor; hyperglycemia