UNIGE document Scientific Article
previous document  unige:4761  next document
add to browser collection

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

Birraux, Jacques
Lasne, Françoise
Nguyen, Tuan H.
show hidden authors show all authors [1 - 9]
Published in Molecular therapy. 2009, vol. 17, no. 10, p. 1754-60
Abstract 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.
Keywords AnimalsAntiviral Agents/pharmacologyBlotting, WesternCell LineCells, CulturedErythropoietin/genetics/physiologyGanciclovir/analogs & derivatives/pharmacologyGene Therapy/methodsGenetic Vectors/geneticsHela CellsHepatocytes/cytology/drug effects/virologyHumansLentivirus/geneticsLiver Diseases/therapyMacaca fascicularisMalePolymerase Chain ReactionSimplexvirus/geneticsThymidine Kinase/genetics/physiologyTransduction, Genetic/methodsViral Proteins/genetics/physiology
PMID: 19568222
Full text
Research group Recherche clinique en chirurgie pédiatrique (886)
(ISO format)
MENZEL, Olivier et al. Biosafety in ex vivo gene therapy and conditional ablation of lentivirally transduced hepatocytes in nonhuman primates. In: Molecular therapy, 2009, vol. 17, n° 10, p. 1754-60. doi: 10.1038/mt.2009.143 https://archive-ouverte.unige.ch/unige:4761

461 hits

0 download


Deposited on : 2010-01-14

Export document
Format :
Citation style :