title: Comparison of murine leukemia virus, human immunodeficiency virus, and adeno-associated virus vectors for gene transfer in multiple myeloma: lentiviral vectors demonstrate a striking capacity to transduce low-proliferating primary tumor cells.
abstract: Genetic modification of primary tumor cells by gene transfer is of major interest to study the role of specific genes in the biology of a given malignancy and to modify tumor cells for therapeutic use. Multiple myeloma (MM) is a low-proliferating cancer, with often less than 1% of the cells in the S phase of the cell cycle. As primary myeloma cells are notoriously difficult to transduce, we conducted a comparison of various viral vectors, known to integrate the transgene of interest into the target genome, for their ability to stably promote the expression of an enhanced green fluorescent protein (EGFP) transgene. We compared three murine leukemia virus-based vectors, differing only in their viral envelope, a human immunodeficiency virus (HIV)-based vector pseudotyped with the vesicular stomatitis virus glycoprotein (VSV-G), and an adeno-associated virus type 2 vector. Transduction characteristics of these vectors were evaluated in human myeloma cell lines and in primary myeloma cells. Unequivocally, we observed that the VSV-G/HIV vector was the most efficient vector for transducing the cell lines and the only one able to transduce primary myeloma cells reproducibly. The mean percentage of transduced primary myeloma cells was 43.6% (range, 16.3-77.6%), with one round of infection at a low multiplicity of infection, including MM cell samples with less than 1% of cells in the S phase. A quantitative polymerase chain reaction assay demonstrated that this more efficient EGFP expression was associated with a higher GFP copy number in the targeted cell. We propose that lentiviral vectors should be used for transduction of nonproliferating primary tumor cells such as myeloma cells.
authors.affiliation: INSERM U475, CHU de Montpellier, 34000 Montpellier, France.
authors.forename: John
authors.initials: J
authors.lastname: De Vos
authors.forename: Claude
authors.initials: C
authors.lastname: Bagnis
authors.forename: Lydie
authors.initials: L
authors.lastname: Bonnafoux
authors.forename: Guilhem
authors.initials: G
authors.lastname: Requirand
authors.forename: Michel
authors.initials: M
authors.lastname: Jourdan
authors.forename: Marie-Christine
authors.initials: MC
authors.lastname: Imbert
authors.forename: Eric
authors.initials: E
authors.lastname: Jourdan
authors.forename: Jean-François
authors.initials: JF
authors.lastname: Rossi
authors.forename: Patrice
authors.initials: P
authors.lastname: Mannoni
authors.forename: Bernard
authors.initials: B
authors.lastname: Klein
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D002455:Cell Division; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D006678:HIV / Q000235:genetics*; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D009052:Leukemia Virus, Murine / Q000235:genetics*; D008562:Membrane Glycoproteins; D051379:Mice; D009101:Multiple Myeloma / Q000235:genetics* / Q000628:therapy*; D014161:Transduction, Genetic; D014407:Tumor Cells, Cultured; D014759:Viral Envelope Proteins
chemical_list: C028391:G protein, vesicular stomatitis virus; D008562:Membrane Glycoproteins; D014759:Viral Envelope Proteins