title: Preclinical study on gene therapy of cervical carcinoma using adeno-associated virus vectors.
abstract: Approximately 90% of cervical carcinomas are causally linked to infections with high-risk human papillomaviruses (HPVs), whose oncogenicity has been assigned to the continued expression of two early genes, E6 and E7. Reversal of the transformed phenotype by inhibiting E6/E7 gene expression therefore provides a suitable goal for future tumor therapy. Using recombinant adeno-associated virus type 2 (AAV-2) vectors, two types of therapeutic genes were expressed in cervical carcinoma cells with the aim of suppressing the E6/E7 oncogenes: (a) antisense E6/E7 and ribozyme genes and (b) the monocyte chemoattractant protein-1 (MCP-1) gene encoding MCP-1. Previous studies have shown that the MCP-1 protein is able to indirectly repress E6/E7 gene expression and is consistently absent in tumorigenic HPV-positive cervical carcinoma cell lines. Here, the effect of these therapeutic genes on tumor formation is analyzed in nude mice after ex vivo gene transfer into a HPV16- or HPV18-positive cervical carcinoma cell line (HeLa or SiHa, respectively). Whereas AAV-2 vector-mediated transfer of antisense or even ribozyme genes did not significantly influence tumor formation from implanted SiHa cells, the transfer and expression of human MCP-1 strongly inhibited the development of tumors derived from either HeLa or SiHa cells. Similar results were also obtained after in vivo delivery of these genes into SiHa-derived tumors. This suggests that transfer of therapeutic genes mediating a systemic effect via recombinant AAV-2 vectors offers a promising approach for the development of gene therapies directed against papillomavirus-induced human cancers.
authors.affiliation: Deutsches Krebsforschungszentrum, Forschungsschwerpunkt Angewandte Tumorvirologie, Heidelberg, Germany.
authors.forename: D
authors.initials: D
authors.lastname: Kunke
authors.forename: D
authors.initials: D
authors.lastname: Grimm
authors.forename: S
authors.initials: S
authors.lastname: Denger
authors.forename: J
authors.initials: J
authors.lastname: Kreuzer
authors.forename: H
authors.initials: H
authors.lastname: Delius
authors.forename: D
authors.initials: D
authors.lastname: Komitowski
authors.forename: J A
authors.initials: JA
authors.lastname: Kleinschmidt
mesh_terms: D000818:Animals; D015152:Blotting, Northern; D002455:Cell Division / Q000187:drug effects; D018932:Chemokine CCL2 / Q000235:genetics; D000229:Dependovirus / Q000235:genetics*; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005455:Fluorescent Antibody Technique; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D006367:HeLa Cells; D006801:Humans; D008264:Macrophages / Q000166:cytology / Q000187:drug effects; D051379:Mice; D008819:Mice, Nude; D008957:Models, Genetic; D016376:Oligonucleotides, Antisense / Q000235:genetics; D009856:Oncogene Proteins, Viral / Q000235:genetics; D050725:Papillomavirus E7 Proteins; D010957:Plasmids / Q000235:genetics; D016337:RNA, Catalytic / Q000235:genetics; D012097:Repressor Proteins; D013997:Time Factors; D014407:Tumor Cells, Cultured; D002583:Uterine Cervical Neoplasms / Q000628:therapy*; D001616:beta-Galactosidase / Q000378:metabolism
chemical_list: D018932:Chemokine CCL2; C068651:E6 protein, Human papillomavirus type 16; D016376:Oligonucleotides, Antisense; D009856:Oncogene Proteins, Viral; D050725:Papillomavirus E7 Proteins; D016337:RNA, Catalytic; D012097:Repressor Proteins; C059731:oncogene protein E7, Human papillomavirus type 16; D001616:beta-Galactosidase