title: Improved Hepatocyte Engraftment After Portal Vein Occlusion in LDL Receptor-Deficient WHHL Rabbits and Lentiviral-Mediated Phenotypic Correction In Vitro.
abstract: Innovative cell-based therapies are considered as alternatives to liver transplantation. Recent progress in lentivirus-mediated hepatocyte transduction has renewed interest in cell therapy for the treatment of inherited liver diseases. However, hepatocyte transplantation is still hampered by inefficient hepatocyte engraftment. We previously showed that partial portal vein embolization (PVE) improved hepatocyte engraftment in a nonhuman primate model. We developed here an ex vivo approach based on PVE and lentiviral-mediated transduction of hepatocytes from normal (New Zealand White, NZW) and Watanabe heritable hyperlipidemic (WHHL) rabbits: the large animal model of familial hypercholesterolemia type IIa (FH). FH is a life-threatening human inherited autosomal disease caused by a mutation in the low-density lipoprotein receptor (LDLR) gene, which leads to severe hypercholesterolemia and premature coronary heart disease. Rabbit hepatocytes were isolated from the resected left liver lobe, and the portal branches of the median lobes were embolized with Histoacryl® glue under radiologic guidance. NZW and WHHL hepatocytes were each labeled with Hoechst dye or transduced with lentivirus expressing GFP under the control of a liver-specific promoter (mTTR, a modified murine transthyretin promoter) and were then immediately transplanted back into donor animals. In our conditions, 65-70% of the NZW and WHHL hepatocytes were transduced. Liver repopulation after transplantation with the Hoechst-labeled hepatocytes was 3.5 ± 2%. It was 1.4 ± 0.6% after transplantation with either the transduced NZW hepatocytes or the transduced WHHL hepatocytes, which was close to that obtained with Hoechst-labeled cells, given the mean transduction efficacy. Transgene expression persisted for at least 8 weeks posttransplantation. Transduction of WHHL hepatocytes with an LDLR-encoding vector resulted in phenotypic correction in vitro as assessed by internalization of fluorescent LDL ligands. In conclusion, our results have applications for the treatment of inherited metabolic liver diseases, such as FH, by transplantation of lentivirally transduced hepatocytes.
authors: INSERM U 972, Univ. Paris-Sud, IFR 93, Bicêtre Hospital , Le Kremlin-Bicêtre , France.
authors: Sylvie
authors: S
authors: Goulinet-Mainot
authors: INSERM U 972, Univ. Paris-Sud, IFR 93, Bicêtre Hospital , Le Kremlin-Bicêtre , France.
authors: Hadrien
authors: H
authors: Tranchart
authors: INSERM U 972, Univ. Paris-Sud, IFR 93, Bicêtre Hospital , Le Kremlin-Bicêtre , France.
authors: Marie-Thérèse
authors: MT
authors: Groyer-Picard
authors: INSERM U 972, Univ. Paris-Sud, IFR 93, Bicêtre Hospital, Le Kremlin-Bicêtre, France; †Department of General Surgery, Univ. Paris-Sud, Antoine Béclère Hospital, Clamart, France.
authors: Panagiotis
authors: P
authors: Lainas
authors: INSERM U 972, Univ. Paris-Sud, IFR 93, Bicêtre Hospital, Le Kremlin-Bicêtre, France; †Department of General Surgery, Univ. Paris-Sud, Antoine Béclère Hospital, Clamart, France.
authors: Papa
authors: P
authors: Saloum Diop
authors: ‡ Department of Animal Surgery, Veterinary School of Nantes , Nantes , France.
authors: Delphine
authors: D
authors: Holopherne
authors: ‡ Department of Animal Surgery, Veterinary School of Nantes , Nantes , France.
authors: Patrick
authors: P
authors: Gonin
authors: ¶ CNRS UMR 8203, Institut Gustave Roussy , Villejuif , France.
authors: Karim
authors: K
authors: Benihoud
authors: # IFR 93, Bicêtre Hospital , Le Kremlin-Bicêtre , France.
authors: Nathalie
authors: N
authors: Ba
authors: ‡ Department of Animal Surgery, Veterinary School of Nantes , Nantes , France.
authors: Olivier
authors: O
authors: Gauthier
authors: INSERM U 972, Univ. Paris-Sud, IFR 93, Bicêtre Hospital, Le Kremlin-Bicêtre, France; †Department of General Surgery, Univ. Paris-Sud, Antoine Béclère Hospital, Clamart, France.
authors: Dominique
authors: D
authors: Franco
authors: * Department of Pathology, Bicêtre Hospital , Le Kremlin-Bicêtre , France.
authors: Catherine
authors: C
authors: Guettier
authors: INSERM U 972, Univ. Paris-Sud, IFR 93, Bicêtre Hospital, Le Kremlin-Bicêtre, France; ††Department of Pediatric Radiology, Bicêtre Hospital, Le Kremlin-Bicêtre, France.
authors: Danièle
authors: D
authors: Pariente
authors: INSERM U 972, Univ. Paris-Sud, IFR 93, Bicêtre Hospital , Le Kremlin-Bicêtre , France.
authors: Anne
authors: A
authors: Weber
authors: INSERM U 972, Univ. Paris-Sud, IFR 93, Bicêtre Hospital, Le Kremlin-Bicêtre, France; †Department of General Surgery, Univ. Paris-Sud, Antoine Béclère Hospital, Clamart, France.
authors: Ibrahim
authors: I
authors: Dagher
authors: ‡‡ INSERM U1064, CHU Hôtel Dieu, Université de Nantes , Nantes , France.
authors: Tuan
authors: T
authors: Huy Nguyen
keywords: Familial hypercholesterolemia; Hepatocyte transplantation; Lentiviral vector; Liver; Portal vein embolization; Rabbit