title: Crry, a complement regulatory protein, modulates renal interstitial disease induced by proteinuria.
abstract: Crry, a complement regulatory protein, modulates renal interstitial disease induced by proteinuria.
  
  
  BACKGROUND
  Recent studies have suggested a role for urinary complement components in mediating tubulointerstitial damage, which is known to have a good correlation with progression of chronic renal diseases. Although accumulating evidence suggests that complement regulatory proteins play an important protective role in glomeruli, their role in renal tubules remains unclear. In order to establish the role of a complement regulatory protein, Crry, in renal tubular injury, we employed a molecular biological approach to block the expression of Crry in tubules of animals with proteinuria induced with puromycin aminonucleoside nephritis (PAN). Methods and Results. Two different antisense oligodeoxynucleotides (ODNs) against Crry were designed and applied to cultured rat mesangial cells in vitro in order to establish their efficacy. Antisense ODN treatment resulted in decreased expression of Crry protein associated with increased sensitivity to complement attack in cell lysis assays compared with control ODN treatment or no treatment (44.7, 1.50, and 1.34%, respectively). Antisense ODNs did not affect the expression of Thy1 as a control, confirming the specificity of our ODNs. In vivo, we performed selective right renal artery perfusion to administer antisense ODNs to the kidney and showed prominent uptake of ODNs by proximal tubular cells. Reduced expression of Crry protein was demonstrated in proximal tubular cells in antisense ODNs-treated kidneys. Normal rats treated with the antisense ODNs did not show any pathological changes. However, in PAN, rats with massive proteinuria showed increased deposition of C3 and C5b-9 in tubules in antisense-treated kidneys, and histological assessment revealed more severe tubulointerstitial injury in antisense-treated animals compared with controls.
  
  
  CONCLUSIONS
  These results establish a pathogenic role for complement in leading to tubulointerstitial injury during proteinuria and, to our knowledge for the first time, show a protective role of a complement regulatory protein, Crry, in renal interstitial disease.
authors.affiliation: Department of Internal Medicine, University of Tokyo School of Medicine, Tokyo, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Hori
authors.forename: K
authors.initials: K
authors.lastname: Yamada
authors.forename: N
authors.initials: N
authors.lastname: Hanafusa
authors.forename: T
authors.initials: T
authors.lastname: Okuda
authors.forename: N
authors.initials: N
authors.lastname: Okada
authors.forename: T
authors.initials: T
authors.lastname: Miyata
authors.forename: W G
authors.initials: WG
authors.lastname: Couser
authors.forename: K
authors.initials: K
authors.lastname: Kurokawa
authors.forename: T
authors.initials: T
authors.lastname: Fujita
authors.forename: M
authors.initials: M
authors.lastname: Nangaku
mesh_terms: D000818:Animals; D000954:Antigens, Surface; D016375:Antisense Elements (Genetics) / Q000493:pharmacokinetics; D002478:Cells, Cultured; D003176:Complement C3 / Q000032:analysis / Q000378:metabolism; D003165:Complement System Proteins / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D005920:Glomerular Mesangium / Q000737:chemistry / Q000166:cytology; D007687:Kidney Tubules, Proximal / Q000166:cytology / Q000502:physiology; D008297:Male; D009395:Nephritis, Interstitial / Q000209:etiology / Q000473:pathology*; D009404:Nephrotic Syndrome / Q000150:complications / Q000473:pathology; D009841:Oligonucleotides / Q000493:pharmacokinetics; D010503:Periodic Acid-Schiff Reaction; D011507:Proteinuria / Q000150:complications / Q000473:pathology*; D051381:Rats; D017208:Rats, Wistar; D011956:Receptors, Cell Surface; D011951:Receptors, Complement / Q000032:analysis / Q000235:genetics*; D012077:Renal Artery; D014162:Transfection; D014746:Vimentin / Q000032:analysis
chemical_list: D000954:Antigens, Surface; D016375:Antisense Elements (Genetics); D003176:Complement C3; C513105:Cr1l protein, rat; D009841:Oligonucleotides; D011956:Receptors, Cell Surface; D011951:Receptors, Complement; D014746:Vimentin; D003165:Complement System Proteins