Suicide gene therapy of sarcoma cell lines using recombinant adeno-associated virus 2 vectors.
Soft-tissue sarcomas are mesenchymal tumors that respond poorly to systemic chemotherapy. Suicide gene therapy may be an alternative treatment strategy. Here we show a high susceptibility of human sarcoma cell lines for recombinant adeno-associated virus 2 (rAAV-2) suicide vectors: connective tissue sarcoma (HS-1), fibrosarcoma (HT-1080), Ewing sarcoma (RD-ES), Askin tumor (SK-N-MC), rhabdomyosarcoma (A-204) and soft-tissue sarcoma (WSKL-1). Several vectors containing the thymidine kinase (TK) gene under the control of either the cytomegalovirus promoter or the elongation-factor 1 alpha (EF1alpha) promoter were cloned and tested. Higher expression levels of the transgene were observed in the sarcoma lines when using the EF1alpha-suicide gene-containing vectors. A complete eradication of rAAV-2-EF1alpha-TK/eGFP (TK/enhanced green fluorescent protein fusion gene)-transduced tumor cells was shown following exposure to ganciclovir (2.5 microg/ml) in vitro, while at this dose level > 90% of mock-transduced tumor cells survived. Xenotransplantation tumor models (intraperitoneal, subcutaneous) for the human sarcoma cell line HS-1 were established in nonobese diabetic/severe-combined immunodeficient mice. Mice transplanted with rAAV-2-EF1alpha-TK/eGFP-transduced and ganciclovir-exposed tumor cells survived > 5 months while in the nontransduced group all mice had died approximately 1 month after inoculation. These data hold promise for further development of rAAV-2-based suicide gene therapy of sarcomas.
Department of Radiation Oncology, Universitätsklinikum Mannheim, University of Heidelberg, Theodor-Kutzer-Ufer 1-3, D-68135, Germany.
Marlon R
MR
Veldwijk
Simone
S
Berlinghoff
Stephanie
S
Laufs
Ulrich R
UR
Hengge
W Jens
WJ
Zeller
Frederik
F
Wenz
Stefan
S
Fruehauf
D000818:Animals; D045744:Cell Line, Tumor; D003587:Cytomegalovirus / Q000235:genetics; D000229:Dependovirus / Q000235:genetics*; D015774:Ganciclovir / Q000627:therapeutic use; D041722:Genes, Transgenic, Suicide / Q000235:genetics*; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D020648:Peptide Elongation Factor 1 / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics; D012509:Sarcoma / Q000188:drug therapy / Q000628:therapy*; D013937:Thymidine Kinase / Q000235:genetics*; D023041:Xenograft Model Antitumor Assays
D020648:Peptide Elongation Factor 1; D013937:Thymidine Kinase; D015774:Ganciclovir