title: Polyploidization without mitosis improves in vivo liver transduction with lentiviral vectors.
abstract: Lentiviral vectors are efficient gene delivery vehicles for therapeutic and research applications. In contrast to oncoretroviral vectors, they are able to infect most nonproliferating cells. In the liver, induction of cell proliferation dramatically improved hepatocyte transduction using all types of retroviral vectors. However, the precise relationship between hepatocyte division and transduction efficiency has not been determined yet. Here we compared gene transfer efficiency in the liver after in vivo injection of recombinant lentiviral or Moloney murine leukemia viral (MoMuLV) vectors in hepatectomized rats treated or not with retrorsine, an alkaloid that blocks hepatocyte division and induces megalocytosis. Partial hepatectomy alone resulted in a similar increase in hepatocyte transduction using either vector. In retrorsine-treated and partially hepatectomized rats, transduction with MoMuLV vectors dropped dramatically. In contrast, we observed that retrorsine treatment combined with partial hepatectomy increased lentiviral transduction to higher levels than hepatectomy alone. Analysis of nuclear ploidy in single cells showed that a high level of transduction was associated with polyploidization. In conclusion, endoreplication could be exploited to improve the efficiency of liver-directed lentiviral gene therapy.
authors.affiliation: INSERM UMR-S 948, Université de Nantes, CHU Hôtel Dieu, 44093 Nantes, France.
authors.forename: Virginie
authors.initials: V
authors.lastname: Pichard
authors.forename: Dominique
authors.initials: D
authors.lastname: Couton
authors.forename: Chantal
authors.initials: C
authors.lastname: Desdouets
authors.forename: Nicolas
authors.initials: N
authors.lastname: Ferry
mesh_terms: D000818:Animals; D002467:Cell Nucleus / Q000821:virology; D049109:Cell Proliferation; D062951:Endoreduplication; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006498:Hepatectomy; D022781:Hepatocytes / Q000187:drug effects; D007150:Immunohistochemistry; D016086:Lentivirus / Q000235:genetics / Q000378:metabolism*; D008099:Liver / Q000187:drug effects / Q000821:virology*; D008297:Male; D008938:Mitosis; D008979:Moloney murine leukemia virus / Q000235:genetics / Q000378:metabolism; D011123:Polyploidy; D011763:Pyrrolizidine Alkaloids / Q000494:pharmacology; D051381:Rats; D011916:Rats, Inbred F344; D011995:Recombination, Genetic; D014161:Transduction, Genetic
chemical_list: D011763:Pyrrolizidine Alkaloids; D049452:Green Fluorescent Proteins; C003300:retrorsine