title: Lentiviral vectors for sustained transgene expression in human bone marrow-derived stromal cells.
abstract: Bone marrow-derived mesenchymal stromal cells (MSCs) have attracted attention as potential platforms for the systemic delivery of therapeutic proteins in vivo following gene transfer using oncogenic retroviruses. However, the major limitations of this strategy include low levels of gene transfer and a general lack of long-term transgene expression. We have investigated the expression of several transgenes in MSCs following HIV-1 lentiviral vector-mediated gene transfer. Vectors containing a variety of strong promoters driving enhanced green fluorescence protein (EGFP) and coral (Discosoma sp.)-derived red fluorescent protein (DsRed) reporter genes pseudotyped with the vesicular stomatitis virus-G (VSV-G) glycoprotein were able to transduce cultured MSCs with high efficiency. Transduction efficiencies and transgene expression levels in MSCs were found to be higher with lentiviral vectors than with a vector based on the murine stem cell virus pseudotyped with VSV-G. Transgene expression was maintained in culture for at least 5 months. HIV-1-based lentiviral vectors were able to transduce clonogenic mesenchymal progenitor cells, which were capable of maintaining transgene expression by their MSC progeny, over several cell divisions and during differentiation into adipocytes, indicating that terminal adipocyte cell differentiation was unaffected by lentivirus-mediated reporter gene transfer. Collectively these results suggest that lentivirus-mediated gene transfer strategies provide an efficient tool for ex vivo modification of MSCs that does not interfere with differentiation.
authors: Gene Therapy Program, Immunology & Parasitology, LSU Health Sciences Center, New Orleans, Louisiana 70112, USA.
authors: Xian-Yang
authors: XY
authors: Zhang
authors: Vincent F
authors: VF
authors: La Russa
authors: Lili
authors: L
authors: Bao
authors: Jay
authors: J
authors: Kolls
authors: Paul
authors: P
authors: Schwarzenberger
authors: Jakob
authors: J
authors: Reiser
mesh_terms: D001854:Bone Marrow Cells / Q000378:metabolism* / Q000821:virology; D004247:DNA / Q000378:metabolism; D017931:DNA Primers / Q000737:chemistry; D005434:Flow Cytometry; D018628:Gene Dosage; D018014:Gene Transfer Techniques; D049452:Green Fluorescent Proteins; D015497:HIV-1 / Q000235:genetics*; D006801:Humans; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D008562:Membrane Glycoproteins; D010957:Plasmids; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D017154:Stromal Cells / Q000378:metabolism* / Q000821:virology; D014161:Transduction, Genetic; D019076:Transgenes / Q000502:physiology*; D014721:Vesicular stomatitis Indiana virus / Q000235:genetics; D014759:Viral Envelope Proteins / Q000378:metabolism; D000097573:Red Fluorescent Protein
chemical_list: D004247:DNA; D017931:DNA Primers; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins; D008562:Membrane Glycoproteins; D014759:Viral Envelope Proteins; D000097573:Red Fluorescent Protein; C028391:G protein, vesicular stomatitis virus