Biosafety in ex vivo gene therapy and conditional ablation of lentivirally transduced hepatocytes in nonhuman primates.

Détails

ID Serval
serval:BIB_C68C44A0855B
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Titre
Biosafety in ex vivo gene therapy and conditional ablation of lentivirally transduced hepatocytes in nonhuman primates.
Périodique
Molecular Therapy
Auteur(s)
Menzel O., Birraux J., Wildhaber B.E., Jond C., Lasne F., Habre W., Trono D., Nguyen T.H., Chardot C.
ISSN
1525-0024 (Electronic)
ISSN-L
1525-0016
Statut éditorial
Publié
Date de publication
2009
Peer-reviewed
Oui
Volume
17
Numéro
10
Pages
1754-1760
Langue
anglais
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov't Publication Status: ppublish
Résumé
Ex vivo gene therapy is an interesting alternative to orthotopic liver transplantation (OLT) for treating metabolic liver diseases. In this study, we investigated its efficacy and biosafety in nonhuman primates. Hepatocytes isolated from liver lobectomy were transduced in suspension with a bicistronic liver-specific lentiviral vector and immediately autotransplanted (SLIT) into three cynomolgus monkeys. The vector encoded cynomolgus erythropoietin (EPO) and the conditional suicide gene herpes simplex virus-thymidine kinase (HSV-TK). Survival of transduced hepatocytes and vector dissemination were evaluated by detecting transgene expression and vector DNA. SLIT was safely performed within a day in all three subjects. Serum EPO and hematocrit rapidly increased post-SLIT and their values returned to baseline within about 1 month. Isoforms of EPO detected in monkeys' sera differed from the physiological renal EPO. In liver biopsies at months 8 and 15, we detected EPO protein, vector mRNA and DNA, demonstrating long-term survival and functionality of transplanted lentivirally transduced hepatocytes. Valganciclovir administration resulted in complete ablation of the transduced hepatocytes. We demonstrated the feasibility and biosafety of SLIT, and the long term (>1 year) functionality of lentivirally transduced hepatocytes in nonhuman primates. The HSV-TK/valganciclovir suicide strategy can increase the biosafety of liver gene therapy protocols by safely and completely ablating transduced hepatocytes on demand.
Mots-clé
Animals, Antiviral Agents/pharmacology, Blotting, Western, Cell Line, Cells, Cultured, Erythropoietin/genetics, Erythropoietin/physiology, Ganciclovir/analogs & derivatives, Ganciclovir/pharmacology, Genetic Therapy/methods, Genetic Vectors/genetics, HeLa Cells, Hepatocytes/cytology, Hepatocytes/drug effects, Humans, Lentivirus/genetics, Liver Diseases/therapy, Macaca fascicularis, Male, Polymerase Chain Reaction, Simplexvirus/genetics, Thymidine Kinase/genetics, Thymidine Kinase/physiology, Transduction, Genetic/methods, Viral Proteins/genetics, Viral Proteins/physiology
Pubmed
Open Access
Oui
Création de la notice
21/02/2015 12:06
Dernière modification de la notice
09/05/2019 1:03
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