Genetically engineered vesicular stomatitis virus in gene therapy: application for treatment of malignant disease.
We report here the generation of recombinant vesicular stomatitis virus (VSV) able to produce the suicide gene product thymidine kinase (TK) or cytokine interleukin 4 (IL-4). In vitro cells infected with the engineered viruses expressed remarkably high levels of biologically active TK or IL-4 and showed no defects in replication compared to the wild-type virus. Recombinant viruses retained their ability to induce potent apoptosis in a variety of cancer cells, while normal cells were evidently more resistant to infection and were completely protected by interferon. Significantly, following direct intratumoral inoculation, VSV expressing either TK or IL-4 exhibited considerably more oncolytic activity against syngeneic breast or melanoma tumors in murine models than did the wild-type virus or control recombinant viruses expressing green fluorescent protein (GFP). Complete regression of a number of tumors was achieved, and increased granulocyte-infiltrating activity with concomitant, antitumor cytotoxic T-cell responses was observed. Aside from discovering greater oncolytic activity following direct intratumoral inoculation, however, we also established that VSV expressing IL-4 or TK, but not GFP, was able to exert enhanced antitumor activity against metastatic disease. Following intravenous administration of the recombinant viruses, immunocompetent BALB/c mice inoculated with mammary adenocarcinoma exhibited prolonged survival against lethal lung metastasis. Our data demonstrate the validity of developing novel types of engineered VSV for recombinant protein production and as a gene therapy vector for the treatment of malignant and other disease.
Fernandez
Marilyn
M
Department of Microbiology and Immunology and Sylvester Comprehensive Cancer Center, University of Miami School of Medicine, Miami, Florida 33136, USA.
Porosnicu
Mercedes
M
Markovic
Dubravka
D
Barber
Glen N
GN
D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D006224:Cricetinae; D004274:DNA, Recombinant / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D005818:Genetic Engineering; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics / Q000502:physiology; D007372:Interferons / Q000494:pharmacology; D015847:Interleukin-4 / Q000032:analysis / Q000235:genetics / Q000276:immunology / Q000627:therapeutic use; D008175:Lung Neoplasms / Q000276:immunology / Q000473:pathology / Q000556:secondary; D015674:Mammary Neoplasms, Animal / Q000276:immunology / Q000473:pathology; D008545:Melanoma / Q000276:immunology / Q000473:pathology; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D009368:Neoplasm Transplantation; D009369:Neoplasms / Q000235:genetics* / Q000473:pathology / Q000628:therapy*; D016019:Survival Analysis; D013937:Thymidine Kinase / Q000235:genetics / Q000378:metabolism / Q000627:therapeutic use; D014407:Tumor Cells, Cultured; D014721:Vesicular stomatitis Indiana virus / Q000235:genetics* / Q000502:physiology
D004274:DNA, Recombinant; D015847:Interleukin-4; D007372:Interferons; D013937:Thymidine Kinase