title: Ex vivo transduced liver progenitor cells as a platform for gene therapy in mice.
abstract: Allogeneic stem cell-based transplants may be limited by allograft rejection, as is seen with conventional organ transplantation. One way to avert such a response is to use autologous stem cells, but that may carry the risk of recurrence of the original disease, particularly in the context of a genetic defect. We investigated the potential for gene modification of autologous stem cells to avoid both problems, using recombinant adenoassociated virus vector expressing human alpha1-antitrypsin in murine liver progenitor cells. We showed that recombinant adenoassociated virus 1 was the most efficient vector for liver progenitor cell transduction among five different serotypes of recombinant adenoassociated virus vectors. Ex vivo infected green fluorescent protein-positive liver progenitor cells from C57BL/6 mice with recombinant adenoassociated virus 1-vector-expressing human alpha1 antitrypsin were transplanted into the liver of monocrotaline-treated and partial-hepatectomized C57BL/6 recipients. Using green fluorescent protein as a donor marker, we were able to determine that at 18 weeks after transplantation, approximately 40% to 50% of the regenerated liver was green fluorescent protein positive. In addition, transgene expression (serum human alpha1-antitrypsin) was sustained for the length of the study (18 weeks after transplantation). Immunostaining revealed approximately 5% to 10% of repopulating liver cells expressing human alpha1-antitrypsin. In conclusion, this study demonstrated the feasibility of long-term engraftment and stability of transgene expression from genetically modified liver progenitor cells with a recombinant adenoassociated virus vector and implies a novel approach to gene therapy for treatment of liver diseases, such as alpha1-antitrypsin deficiency.
authors.affiliation: Department of Pharmaceutics, University of Florida, Gainesville, FL 32610, USA. shsong@ufl.edu
authors.forename: Sihong
authors.initials: S
authors.lastname: Song
authors.forename: Rafal P
authors.initials: RP
authors.lastname: Witek
authors.forename: Yuanqing
authors.initials: Y
authors.lastname: Lu
authors.forename: Young-Kook
authors.initials: YK
authors.lastname: Choi
authors.forename: Donghang
authors.initials: D
authors.lastname: Zheng
authors.forename: Marda
authors.initials: M
authors.lastname: Jorgensen
authors.forename: Chengwen
authors.initials: C
authors.lastname: Li
authors.forename: Terence R
authors.initials: TR
authors.lastname: Flotte
authors.forename: Byron E
authors.initials: BE
authors.lastname: Petersen
mesh_terms: D000818:Animals; D005260:Female; D015316:Genetic Therapy / Q000379:methods*; D049452:Green Fluorescent Proteins; D006498:Hepatectomy; D008099:Liver / Q000166:cytology; D008107:Liver Diseases / Q000628:therapy*; D008164:Luminescent Proteins / Q000235:genetics; D051379:Mice; D008810:Mice, Inbred C57BL; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000166:cytology / Q000502:physiology*; D014161:Transduction, Genetic; D019076:Transgenes; D000515:alpha 1-Antitrypsin / Q000235:genetics; D019896:alpha 1-Antitrypsin Deficiency / Q000628:therapy*
chemical_list: D008164:Luminescent Proteins; D000515:alpha 1-Antitrypsin; D049452:Green Fluorescent Proteins