Gastroenterology

Gastroenterology

Volume 119, Issue 6, December 2000, Pages 1720-1730
Gastroenterology

Liver, Pancreas, and Biliary Tract
Correction of liver disease by hepatocyte transplantation in a mouse model of progressive familial intrahepatic cholestasis,☆☆

https://doi.org/10.1053/gast.2000.20222Get rights and content

Abstract

Background & Aims: Patients with progressive familial intrahepatic cholestasis (PFIC) type 3 have a mutation in the MDR3 gene, encoding the hepatocanalicular phospholipid translocator. In general, liver failure develops within the first decade of life in these patients. Previous studies have shown that in the mdr2-knockout mouse, the animal model for this disease, the absence of phospholipids in bile causes chronic bile salt–induced damage to hepatocytes. We aimed to test the efficacy of hepatocyte transplantation and liver repopulation in this disease model. Methods: Transgenic MDR3-expressing hepatocytes as well as normal mdr2(+/+) hepatocytes were transplanted in mdr2(−/−) mice, and liver repopulation was assessed by immunohistochemistry and measurement of biliary lipid secretion. Results: Transplanted hepatocytes partially repopulated the liver, restored phospholipid secretion, and diminished liver pathology. Repopulation was stronger when hepatocellular damage was enhanced by a bile salt–supplemented diet. After 1 year, however, these animals developed multiple hepatic tumors, and biliary phospholipid secretion decreased. In transplanted animals receiving a control diet, repopulation was slower but eventually remained stable at 21%, while liver pathology was completely abrogated and tumor formation was prevented. Conclusions: These results suggest that moderate liver pathology is a safe condition for the induction of effective hepatocyte repopulation and that this therapy is potentially applicable to patients with PFIC type 3.

GASTROENTEROLOGY 2000;119:1720-1730

Section snippets

Experimental protocol

Hepatocyte transplantation was performed in male mdr2(−/−) mice, the generation of which has been described previously.11 For donor mice, a transgenic A63 mouse strain that expresses the homologous human MDR3 gene behind an albumin promoter and was bred against the identical FVB background was used.13, 14 Compared with mdr2(+/+) control mice, phospholipid secretion in A63 mice is 180%.13, 14 MDR3-expressing hepatocytes were isolated from 3-month-old male A63 mice by ex vivo collagenase

Results

We tested the hypothesis that chronic damage to hepatocytes in a condition of defective biliary phospholipid secretion, i.e., in patients with PFIC type 3 and in mdr2(−/−) mice, causes a state of regenerative proliferation. This was done by immunostaining for Ki67 in samples from a PFIC type 3 patient and by staining for PCNA in sections from mdr2(−/−) livers (both markers stain nuclei of proliferating hepatocytes). Indeed, in livers from both the PFIC type 3 patients (Figure 1A) and mdr2(−/−)

Discussion

Lack of biliary phospholipid secretion leads to liver disease in both mice19 and humans.12 This is caused by the cytotoxic action of high concentrations of bile salts in the biliary tree. We hypothesized that this chronic damage to parenchymal cells leads to a continuous state of regenerative proliferation that makes these livers amenable to repopulation by transplanted hepatocytes. Our results now show that, at least for the mouse model, this hypothesis is correct. From the extensive

Acknowledgements

The authors thank Piet Borst and Bert Groen for critically reading the manuscript, Wilfried Meun for assistance with microphotography, Jos Mulder for assistance with PCNA immunohistochemistry, and Isabel Wesdorp for performing reverse-transcription polymerase chain reactions.

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  • Cited by (0)

    Address requests for reprints to: Ronald P. J. Oude Elferink, Ph.D., Department of Gastroenterology and Hepatology Fo-116, Meibergdreef 9, 1105 AZ Amsterdam, The Netherlands. e-mail: [email protected]; fax: (31) 20-5669190.

    ☆☆

    Supported by project no. 902-23-097 from the Dutch Association for Scientific Research.

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