Tumor-selective gene transduction and cell killing with an oncotropic autonomous parvovirus-based vector.
A recombinant MVMp of the fibrotropic strain of minute virus of mice (MVMp) expressing the chloramphenicol acetyltransferase reporter gene was used to infect a series of biologically relevant cultured cells, normal or tumor-derived, including normal melanocytes versus melanoma cells, normal mammary epithelial cells versus breast adenocarcinoma cells, and normal neurons or astrocytes versus glioma cells. As a reference cell system we used normal human fibroblasts versus the SV40-transformed fibroblast cell line NB324K. After infection, we observed good expression of the reporter gene in the different tumor cell types, but only poor expression if any in the corresponding normal cells. We also constructed a recombinant MVMp expressing the green fluorescent protein reporter gene and assessed by flow cytometry the efficiency of gene transduction into the different target cells. At a multiplicity of infection of 30, we observed substantial transduction of the gene into most of the tumor cell types tested, but only marginal transduction into normal cells under the same experimental conditions. Finally, we demonstrated that a recombinant MVMp expressing the herpes simplex virus thymidine kinase gene can, in vitro, cause efficient killing of most tumor cell types in the presence of ganciclovir, whilst affecting normal proliferating cells only marginally if at all. However, in the same experimental condition, breast tumor cells appeared to be resistant to GCV-mediated cytotoxicity, possibly because these cells are not susceptible to the bystander effect. Our data suggest that MVMp-based vectors could prove useful as selective vehicles for anticancer gene therapy, particularly for in vivo delivery of cytotoxic effector genes into tumor cells.
Dupont
F
F
Laboratoire d'Investigation Clinique et d'Oncologie Expérimentale, Unité d'Oncologie Moléculaire, Belgium.
Avalosse
B
B
Karim
A
A
Mine
N
N
Bosseler
M
M
Maron
A
A
Van den Broeke
A V
AV
Ghanem
G E
GE
Burny
A
A
Zeicher
M
M
D000230:Adenocarcinoma / Q000628:therapy; D000818:Animals; D001943:Breast Neoplasms / Q000628:therapy; D005260:Female; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000008:administration & dosage*; D005910:Glioma / Q000628:therapy; D018259:Herpesvirus 1, Human / Q000201:enzymology; D006801:Humans; D008544:Melanocytes; D008545:Melanoma / Q000628:therapy; D008915:Minute Virus of Mice / Q000235:genetics*; D009369:Neoplasms / Q000628:therapy*; D051381:Rats; D013937:Thymidine Kinase / Q000235:genetics; D014162:Transfection / Q000379:methods*; D014407:Tumor Cells, Cultured
D013937:Thymidine Kinase