title: Urinary CCN2 (CTGF) as a possible predictor of diabetic nephropathy: preliminary report.
abstract: BACKGROUND
  It is currently impossible to reliably predict which diabetic patients will develop nephropathy and progress to kidney failure. Microalbuminuria, often regarded as a predictor of overt diabetic renal disease is, in fact, an indicator of established glomerular damage. We have shown that glomerular expression of the prosclerotic cytokine CCN2 (CTGF) is greatly up-regulated early in experimental and in human diabetes and mesangial cell exposure to CCN2 increases its production of extracellular matrix (ECM) molecules responsible for glomerulosclerosis. As an early marker, we therefore investigated the presence of CCN2 in urine and the relationship to diabetes and/or renal disease in an experimental model of diabetes and in a limited patient population.
  
  
  METHODS
  Urine samples from (1) healthy rats, (2) rats made diabetic by streptozotocin (STZ), (3) healthy human volunteers, (4) diabetic patients with renal disease, and (5) diabetic patients without renal disease were examined by Western blotting and/or enzyme-linked immunosorbent assay (ELISA) for qualitative and quantitative analysis of the of CCN2.
  
  
  RESULTS
  Low levels of urinary CCN2 were present in healthy, control rats, but were increased approximately sevenfold overall in STZ-diabetic animals. CCN2 levels were the highest at week 3 of diabetes, then decreased with time, but remained significantly elevated over controls even after 32 weeks. Consistently low levels of urinary CCN2 were also detected in healthy volunteers (mean value, 7.1 CCN2/mg creatinine). However, levels were elevated approximately sixfold in the majority of diabetic patients with nephropathy. A small number of the diabetic patients not yet exhibiting evidence of renal involvement demonstrated CCN2 urinary levels that were ninefold greater than controls. The remaining normoalbuminuric diabetic patients demonstrated CCN2 levels indistinguishable from those of healthy volunteers. Analysis by Western blotting confirmed the identity of the urinary CCN2. A molecular species equivalent to full-length CCN2 (37/39 kD doublet) was present in healthy controls. In contrast, the nephropathic group demonstrated multiple CCN2 bands.
  
  
  CONCLUSIONS
  These findings support our hypothesis that CCN2 is up-regulated early in the evolution of glomerulosclerosis, including that of diabetes. We contend that urinary CCN2 may both stage nephropathy and predict those patients who are destined for progressive glomerulosclerosis and end-stage renal disease (ESRD). Cross-sectional and prospective studies of larger, well-defined diabetic patients groups will be required to prove this hypothesis, and are ongoing.
authors.affiliation: Department of Medicine, Division of Nephrology and Hypertension, Henry Ford Health Center, Detroit, Michigan, USA. bruce_riser@baxter.com
authors.forename: Bruce L
authors.initials: BL
authors.lastname: Riser
authors.forename: Pedro
authors.initials: P
authors.lastname: Cortes
authors.forename: Mark
authors.initials: M
authors.lastname: DeNichilo
authors.forename: Poornima V
authors.initials: PV
authors.lastname: Deshmukh
authors.forename: Parminder S
authors.initials: PS
authors.lastname: Chahal
authors.forename: Ali K
authors.initials: AK
authors.lastname: Mohammed
authors.forename: Jerry
authors.initials: J
authors.lastname: Yee
authors.forename: Dorothy
authors.initials: D
authors.lastname: Kahkonen
mesh_terms: D000818:Animals; D015415:Biomarkers; D055513:Connective Tissue Growth Factor; D003921:Diabetes Mellitus, Experimental / Q000652:urine; D003928:Diabetic Nephropathies / Q000175:diagnosis* / Q000652:urine*; D006801:Humans; D017874:Immediate-Early Proteins / Q000652:urine*; D036341:Intercellular Signaling Peptides and Proteins / Q000652:urine*; D008297:Male; D010865:Pilot Projects; D011237:Predictive Value of Tests; D051381:Rats; D011916:Rats, Inbred F344
chemical_list: D015415:Biomarkers; C526317:CCN2 protein, human; C526316:CCN2 protein, rat; D017874:Immediate-Early Proteins; D036341:Intercellular Signaling Peptides and Proteins; D055513:Connective Tissue Growth Factor