Limits...
COMMD1-deficient dogs accumulate copper in hepatocytes and provide a good model for chronic hepatitis and fibrosis.

Favier RP, Spee B, Schotanus BA, van den Ingh TS, Fieten H, Brinkhof B, Viebahn CS, Penning LC, Rothuizen J - PLoS ONE (2012)

Bottom Line: Both HGF and TGF-β1 gene expression peaked at 24 months, and thereafter decreased gradually.Both STAT3 and c-MET showed an increased time-dependent activation.Therefore they represent a genetically-defined large animal model to test clinical applicability of new therapeutics developed in rodent models.

View Article: PubMed Central - PubMed

Affiliation: Department of Clinical Sciences of Companion Animals, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands. r.p.favier@uu.nl

ABSTRACT
New therapeutic concepts developed in rodent models should ideally be evaluated in large animal models prior to human clinical application. COMMD1-deficiency in dogs leads to hepatic copper accumulation and chronic hepatitis representing a Wilson's disease like phenotype. Detailed understanding of the pathogenesis and time course of this animal model is required to test its feasibility as a large animal model for chronic hepatitis. In addition to mouse models, true longitudinal studies are possible due to the size of these dogs permitting detailed analysis of the sequence of events from initial insult to final cirrhosis. Therefore, liver biopsies were taken each half year from five new born COMMD1-deficient dogs over a period of 42 months. Biopsies were used for H&E, reticulin, and rubeanic acid (copper) staining. Immunohistochemistry was performed on hepatic stellate cell (HSC) activation marker (alpha-smooth muscle actin, α-SMA), proliferation (Ki67), apoptosis (caspase-3), and bile duct and liver progenitor cell (LPC) markers keratin (K) 19 and 7. Quantitative RT-PCR and Western Blots were performed on gene products involved in the regenerative and fibrotic pathways. Maximum copper accumulation was reached at 12 months of age, which coincided with the first signs of hepatitis. HSCs were activated (α-SMA) from 18 months onwards, with increasing reticulin deposition and hepatocytic proliferation in later stages. Hepatitis and caspase-3 activity (first noticed at 18 months) increased over time. Both HGF and TGF-β1 gene expression peaked at 24 months, and thereafter decreased gradually. Both STAT3 and c-MET showed an increased time-dependent activation. Smad2/3 phosphorylation, indicative for fibrogenesis, was present at all time-points. COMMD1-deficient dogs develop chronic liver disease and cirrhosis comparable to human chronic hepatitis, although at much higher pace. Therefore they represent a genetically-defined large animal model to test clinical applicability of new therapeutics developed in rodent models.

Show MeSH

Related in: MedlinePlus

TGF-β1 pathway.(A) Gene-expression profiling. Interaction plots of Q-PCR data of important mediators of fibrogenesis in a time-dependent copper-induced hepatitis in five COMMD1-deficient dogs. Q-PCR results were normalized against the expression of six control dogs (one to three years of age). Linear mixed-effect modeling was used; * significant difference corrected for multiple testing. (B) Western blot analysis on the activation of TGF-beta signalling (total Smad2 and phosphorylated Smad2 (Ser727)) of five COMMD1-deficient dogs at 12, 30, and 42 months. β-actin (ACTB) served as loading control.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC3412840&req=5

pone-0042158-g002: TGF-β1 pathway.(A) Gene-expression profiling. Interaction plots of Q-PCR data of important mediators of fibrogenesis in a time-dependent copper-induced hepatitis in five COMMD1-deficient dogs. Q-PCR results were normalized against the expression of six control dogs (one to three years of age). Linear mixed-effect modeling was used; * significant difference corrected for multiple testing. (B) Western blot analysis on the activation of TGF-beta signalling (total Smad2 and phosphorylated Smad2 (Ser727)) of five COMMD1-deficient dogs at 12, 30, and 42 months. β-actin (ACTB) served as loading control.

Mentions: The mRNA levels of TGF-β1 and both its receptors increased significantly starting at 24 months and decreased gradually afterwards (Figure 2A). The increase in relative mRNA levels of HGF and its receptor c-MET was sharp and strong (15 and 11-fold, respectively) occurring at 24 months of age in all dogs (Figure 3A). These levels decreased gradually over time and returned to basal levels at 36 months of age.


COMMD1-deficient dogs accumulate copper in hepatocytes and provide a good model for chronic hepatitis and fibrosis.

Favier RP, Spee B, Schotanus BA, van den Ingh TS, Fieten H, Brinkhof B, Viebahn CS, Penning LC, Rothuizen J - PLoS ONE (2012)

TGF-β1 pathway.(A) Gene-expression profiling. Interaction plots of Q-PCR data of important mediators of fibrogenesis in a time-dependent copper-induced hepatitis in five COMMD1-deficient dogs. Q-PCR results were normalized against the expression of six control dogs (one to three years of age). Linear mixed-effect modeling was used; * significant difference corrected for multiple testing. (B) Western blot analysis on the activation of TGF-beta signalling (total Smad2 and phosphorylated Smad2 (Ser727)) of five COMMD1-deficient dogs at 12, 30, and 42 months. β-actin (ACTB) served as loading control.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3412840&req=5

pone-0042158-g002: TGF-β1 pathway.(A) Gene-expression profiling. Interaction plots of Q-PCR data of important mediators of fibrogenesis in a time-dependent copper-induced hepatitis in five COMMD1-deficient dogs. Q-PCR results were normalized against the expression of six control dogs (one to three years of age). Linear mixed-effect modeling was used; * significant difference corrected for multiple testing. (B) Western blot analysis on the activation of TGF-beta signalling (total Smad2 and phosphorylated Smad2 (Ser727)) of five COMMD1-deficient dogs at 12, 30, and 42 months. β-actin (ACTB) served as loading control.
Mentions: The mRNA levels of TGF-β1 and both its receptors increased significantly starting at 24 months and decreased gradually afterwards (Figure 2A). The increase in relative mRNA levels of HGF and its receptor c-MET was sharp and strong (15 and 11-fold, respectively) occurring at 24 months of age in all dogs (Figure 3A). These levels decreased gradually over time and returned to basal levels at 36 months of age.

Bottom Line: Both HGF and TGF-β1 gene expression peaked at 24 months, and thereafter decreased gradually.Both STAT3 and c-MET showed an increased time-dependent activation.Therefore they represent a genetically-defined large animal model to test clinical applicability of new therapeutics developed in rodent models.

View Article: PubMed Central - PubMed

Affiliation: Department of Clinical Sciences of Companion Animals, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands. r.p.favier@uu.nl

ABSTRACT
New therapeutic concepts developed in rodent models should ideally be evaluated in large animal models prior to human clinical application. COMMD1-deficiency in dogs leads to hepatic copper accumulation and chronic hepatitis representing a Wilson's disease like phenotype. Detailed understanding of the pathogenesis and time course of this animal model is required to test its feasibility as a large animal model for chronic hepatitis. In addition to mouse models, true longitudinal studies are possible due to the size of these dogs permitting detailed analysis of the sequence of events from initial insult to final cirrhosis. Therefore, liver biopsies were taken each half year from five new born COMMD1-deficient dogs over a period of 42 months. Biopsies were used for H&E, reticulin, and rubeanic acid (copper) staining. Immunohistochemistry was performed on hepatic stellate cell (HSC) activation marker (alpha-smooth muscle actin, α-SMA), proliferation (Ki67), apoptosis (caspase-3), and bile duct and liver progenitor cell (LPC) markers keratin (K) 19 and 7. Quantitative RT-PCR and Western Blots were performed on gene products involved in the regenerative and fibrotic pathways. Maximum copper accumulation was reached at 12 months of age, which coincided with the first signs of hepatitis. HSCs were activated (α-SMA) from 18 months onwards, with increasing reticulin deposition and hepatocytic proliferation in later stages. Hepatitis and caspase-3 activity (first noticed at 18 months) increased over time. Both HGF and TGF-β1 gene expression peaked at 24 months, and thereafter decreased gradually. Both STAT3 and c-MET showed an increased time-dependent activation. Smad2/3 phosphorylation, indicative for fibrogenesis, was present at all time-points. COMMD1-deficient dogs develop chronic liver disease and cirrhosis comparable to human chronic hepatitis, although at much higher pace. Therefore they represent a genetically-defined large animal model to test clinical applicability of new therapeutics developed in rodent models.

Show MeSH
Related in: MedlinePlus