title: PDGF-A promoter and enhancer elements provide efficient and selective antineoplastic gene therapy in multiple cancer types.
abstract: Development of antineoplastic gene therapies is impaired by a paucity of transcription control elements with efficient, cancer cell-specific activity. We investigated the utility of promoter (AChP) and 5'-distal enhancer (ACE66) elements from the platelet-derived growth factor-A (PDGF-A) gene, which are hyperactive in many human cancers. Efficacy of these elements was tested in multiple tumor cell lines, both in cell culture and as tumor explants in athymic nude mice. Plasmid and viral vectors were constructed with the AChP promoter alone or in fusion with three copies of the ACE66 enhancer for expression of the prototype suicide gene, thymidine kinase (TK). ACE/AChP and AChP cassettes elicited ganciclovir (GCV)-induced cytotoxicity in multiple tumor cell lines. The ACE enhancer element also exhibited synergism with placental and liver-specific promoter elements. An adenovirus containing the AChP-TK cassette produced striking increases in GCV sensitivity in cultured tumor cell lines, as well as GCV-induced regression of U87 MG glioblastoma explants in vivo. TK expression was distributed throughout tumors receiving the therapeutic virus, whereas TdT-mediated dUTP nick end labeling (TUNEL) analysis revealed numerous regions undergoing apoptosis. Vascularization and reticulin fiber networks were less pronounced in virus-GCV-treated tumors, suggesting that both primary and stromal cell types may have been targeted. These studies provide proof-of-principle for utility of the PDGF-A promoter and ACE66 enhancer in antineoplastic gene therapy for a diverse group of human cancers.
authors.affiliation: Department of Molecular and Biomedical Pharmacology, College of Medicine, University of Kentucky, Lexington, KY 40536-0298, USA.
authors.forename: A
authors.initials: A
authors.lastname: Mishra
authors.forename: A K
authors.initials: AK
authors.lastname: Ormerod
authors.forename: M L
authors.initials: ML
authors.lastname: Cibull
authors.forename: B T
authors.initials: BT
authors.lastname: Spear
authors.forename: S D
authors.initials: SD
authors.lastname: Kraner
authors.forename: D M
authors.initials: DM
authors.lastname: Kaetzel
mesh_terms: D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects; D045744:Cell Line, Tumor; D003131:Combined Modality Therapy; D004742:Enhancer Elements, Genetic; D005260:Female; D015774:Ganciclovir / Q000494:pharmacology; D015316:Genetic Therapy; D006801:Humans; D051379:Mice; D009369:Neoplasms / Q000235:genetics / Q000378:metabolism / Q000628:therapy*; D010982:Platelet-Derived Growth Factor / Q000235:genetics* / Q000627:therapeutic use; D011401:Promoter Regions, Genetic; D013937:Thymidine Kinase / Q000096:biosynthesis / Q000235:genetics; D023041:Xenograft Model Antitumor Assays
chemical_list: D000998:Antiviral Agents; D010982:Platelet-Derived Growth Factor; C075278:platelet-derived growth factor A; D013937:Thymidine Kinase; D015774:Ganciclovir