Hepatic repopulation with stably transduced conditionally immortalized hepatocytes in the Gunn rat

Yujo Kawashita, Chandan Guha, Rituparna Moitra, Xia Wang, Ira J. Fox, Jayanta Roy-Chowdhury, Namita Roy-Chowdhury

Research output: Contribution to journalArticle

18 Scopus citations

Abstract

Background/Aims: Conditionally immortalized hepatocytes offer a renewable source of hepatocytes, but although preparative maneuvers have been developed for hepatic repopulation with primary hepatocytes, extensive proliferation of transplanted immortalized hepatocytes has not been accomplished heretofore. Our aim was to achieve ex vivo gene therapy of uridinediphosphoglucuronate glucuronosyltransferase-1A1 (UGT1A1)-deficient jaundiced Gunn rats (model of Crigler-Najjar syndrome type-1) by hepatic repopulation with genetically modified and conditionally immortalized hepatocytes. Methods: Gunn rat hepatocytes were conditionally immortalized by stable transduction with a thermolabile mutant simian virus 40 T-antigen (tsTagA58) and further transduced with UGT1A1. These hepatocytes proliferate at 33°C, but at 37°C the tsTagA58 is degraded and the cells become quiescent. The cells were transplanted into Gunn rat livers after preparative hepatic irradiation (50 Gy) and 66% hepatectomy. Results: The engrafted UGT1A1-positive immortalized hepatocytes replaced ∼80% of the host hepatocytes in 20 weeks, leading to normalization of hyperbilirubinemia. Liver histology, and serum albumin and alanine aminotransferase levels remained normal. Conclusions: We achieved complete cure of hyperbilirubinemia in Gunn rats by ex vivo gene therapy via genetically modified and conditionally immortalized hepatocytes.

Original languageEnglish (US)
Pages (from-to)99-106
Number of pages8
JournalJournal of Hepatology
Volume49
Issue number1
DOIs
StatePublished - Jul 1 2008

Keywords

  • Crigler-Najjar syndrome
  • Gene therapy
  • Hepatic irradiation
  • Hepatic repopulation
  • Immortalized hepatocytes

ASJC Scopus subject areas

  • Hepatology

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