Evaluation of the accuracy of exchangeable copper and relative exchangeable copper (REC) in a mouse model of Wilson's disease

J Trace Elem Med Biol. 2018 Dec;50:652-657. doi: 10.1016/j.jtemb.2018.06.013. Epub 2018 Jun 21.


Wilson's disease (WD) is caused by mutations in the ATP7B gene responsible for a toxic copper overload mainly in the liver and the central nervous system. Phenotypic heterogeneity may challenge the diagnostic confirmation. Exchangeable copper (CuEXC) has recently been proposed as a new marker of WD, and its ratio to the total serum copper (Cus), Relative Exchangeable Copper (REC = CuEXC/Cus), as a diagnostic marker. This study aimed to investigate whether this could be confirmed in Atp7b-/- mice, an engineered WD animal model. Atp7b-/- (n = 137) and wild type (WT; n = 101) mice were investigated under the same conditions at 6-8, 20, 39, or 50 weeks of age. Twenty-four Atp7b-/- mice received D-penicillamine treatment from 39 to 50 weeks of age. Serum and liver [histology and intrahepatic copper (IHCu)] data were evaluated. In the WT group, all serum and liver data were normal. Atp7b-/- livers developed a chronic injury from isolated moderate inflammation (6-8 weeks: 16/33 = 48%) to inflammatory fibrosis with cirrhosis (50 weeks: 25/25 = 100% and 16/25 = 64% respectively). Cus and CuEXC increased until week 39, whereas IHCu and REC were stable with increasing age and much higher than in WT mice (mean ± SD: 669 ± 269 vs. 13 ± 3 μg/g dry liver and 39 ± 12 vs. 11 ± 3%, respectively). A threshold value of 20% for REC provided a diagnostic sensitivity and specificity of 100%, regardless of sex, age, or the use of D-penicillamine. Eleven weeks of 100 mg/kg D-penicillamine reduced liver fibrosis (p = 0.001), IHCu (p = 0.026) and CuEXC (p = 0.175). In conclusion, this study confirms REC as a WD diagnostic marker in a mouse model of chronic liver disease caused by copper overload. Further studies are needed to assess the usefulness of CuEXC to monitor the evolution of WD, particularly during treatment.

Keywords: Atp7b(-/-) mice; Cirrhosis; Diagnostic test; Exchangeable copper; REC.

MeSH terms

  • Adenosine Triphosphatases / blood
  • Alanine / blood
  • Animals
  • Aspartate Aminotransferases / blood
  • Bilirubin / blood
  • Biomarkers / blood
  • Copper / blood*
  • Copper-Transporting ATPases / genetics
  • Disease Models, Animal
  • Fibrosis / blood
  • Fibrosis / genetics
  • Hepatolenticular Degeneration / blood*
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Mutation / genetics


  • Biomarkers
  • Copper
  • Aspartate Aminotransferases
  • Adenosine Triphosphatases
  • Atp7b protein, mouse
  • Copper-Transporting ATPases
  • Alanine
  • Bilirubin