title: Podocyte vascular endothelial growth factor (Vegf₁₆₄) overexpression causes severe nodular glomerulosclerosis in a mouse model of type 1 diabetes.
abstract: OBJECTIVE
  The pathogenic role of excessive vascular endothelial growth factor (VEGF)-A in diabetic nephropathy has not been defined. We sought to test whether increased podocyte VEGF-A signalling determines the severity of diabetic glomerulopathy.
  
  
  METHODS
  Podocyte-specific, doxycycline-inducible Vegf₁₆₄ (the most abundant Vegfa isoform) overexpressing adult transgenic mice were made diabetic with low doses of streptozotocin and examined 12 weeks after onset of diabetes. We studied diabetic and non-diabetic transgenic mice fed a standard or doxycycline-containing diet. VEGF-A and albuminuria were measured by ELISA, creatinine was measured by HPLC, renal morphology was examined by light and electron microscopy, and gene expression was assessed by quantitative PCR, immunoblotting and immunohistochemistry.
  
  
  RESULTS
  Podocyte Vegf₁₆₄ overexpression in our mouse model of diabetes resulted in advanced diabetic glomerulopathy, characterised by Kimmelstiel-Wilson-like nodular glomerulosclerosis, microaneurysms, mesangiolysis, glomerular basement membrane thickening, podocyte effacement and massive proteinuria associated with hyperfiltration. It also led to increased VEGF receptor 2 and semaphorin3a levels, as well as nephrin and matrix metalloproteinase-2 downregulation, whereas circulating VEGF-A levels were similar to those in control diabetic mice.
  
  
  CONCLUSIONS
  Collectively, these data demonstrate that increased podocyte Vegf₁₆₄ signalling dramatically worsens diabetic nephropathy in a streptozotocin-induced mouse model of diabetes, resulting in nodular glomerulosclerosis and massive proteinuria. This suggests that local rather than systemic VEGF-A levels determine the severity of diabetic nephropathy and that semaphorin3a signalling and matrix metalloproteinase-2 dysregulation are mechanistically involved in severe diabetic glomerulopathy.
authors.affiliation: Department of Pediatrics, Yale University School of Medicine, 333 Cedar St, P.O. Box 208064, New Haven, CT 06520-8064, USA.
authors.forename: D
authors.initials: D
authors.lastname: Veron
authors.forename: C A
authors.initials: CA
authors.lastname: Bertuccio
authors.forename: A
authors.initials: A
authors.lastname: Marlier
authors.forename: K
authors.initials: K
authors.lastname: Reidy
authors.forename: A M
authors.initials: AM
authors.lastname: Garcia
authors.forename: J
authors.initials: J
authors.lastname: Jimenez
authors.forename: H
authors.initials: H
authors.lastname: Velazquez
authors.forename: M
authors.initials: M
authors.lastname: Kashgarian
authors.forename: G W
authors.initials: GW
authors.lastname: Moeckel
authors.forename: A
authors.initials: A
authors.lastname: Tufro
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002851:Chromatography, High Pressure Liquid; D003404:Creatinine / Q000097:blood / Q000652:urine; D003922:Diabetes Mellitus, Type 1 / Q000235:genetics / Q000378:metabolism*; D003928:Diabetic Nephropathies / Q000209:etiology* / Q000235:genetics / Q000378:metabolism*; D004195:Disease Models, Animal; D004797:Enzyme-Linked Immunosorbent Assay; D007150:Immunohistochemistry; D051379:Mice; D008822:Mice, Transgenic; D046529:Microscopy, Electron, Transmission; D050199:Podocytes / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D040121:Semaphorin-3A / Q000235:genetics / Q000378:metabolism; D053719:Tandem Mass Spectrometry; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics / Q000378:metabolism*
chemical_list: D040121:Semaphorin-3A; D042461:Vascular Endothelial Growth Factor A; D003404:Creatinine