title: AAVPG: a vigilant vector where transgene expression is induced by p53.
abstract: Using p53 to drive transgene expression from viral vectors may provide on demand expression in response to physiologic stress, such as hypoxia or DNA damage. Here we introduce AAVPG, an adeno-associated viral (AAV) vector where a p53-responsive promoter, termed PG, is used to control transgene expression. In vitro assays show that expression from the AAVPG-luc vector was induced specifically in the presence of functional p53 (1038±202 fold increase, p<0.001). The AAVPG-luc vector was an effective biosensor of p53 activation in response to hypoxia (4.48±0.6 fold increase in the presence of 250µM CoCl2, p<0.001) and biomechanical stress (2.53±0.4 fold increase with stretching, p<0.05). In vivo, the vigilant nature of the AAVPG-luc vector was revealed after treatment of tumor-bearing mice with doxorubicin (pre-treatment, 3.4×10(5)±0.43×10(5)photons/s; post-treatment, 6.6×10(5)±2.1×10(5)photons/s, p<0.05). These results indicate that the AAVPG vector is an interesting option for detecting p53 activity both in vitro and in vivo.
authors.affiliation: Viral Vector Laboratory, Laboratory of Genetics and Molecular Cardiology/LIM13, Heart Institute, University of São Paulo School of Medicine, São Paulo, SP, Brazil.
authors.forename: Marcio C
authors.initials: MC
authors.lastname: Bajgelman
authors.forename: Ruan F V
authors.initials: RF
authors.lastname: Medrano
authors.forename: Anna Carolina P V
authors.initials: AC
authors.lastname: Carvalho
authors.forename: Bryan E
authors.initials: BE
authors.lastname: Strauss
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000008:administration & dosage; D015374:Biosensing Techniques / Q000379:methods; D000229:Dependovirus / Q000235:genetics*; D004195:Disease Models, Animal; D004317:Doxorubicin / Q000008:administration & dosage; D005260:Female; D015870:Gene Expression / Q000187:drug effects*; D017930:Genes, Reporter; D005822:Genetic Vectors; D000860:Hypoxia; D008156:Luciferases / Q000096:biosynthesis / Q000235:genetics; D051379:Mice; D008810:Mice, Inbred C57BL; D009369:Neoplasms / Q000188:drug therapy; D011401:Promoter Regions, Genetic; D013312:Stress, Physiological; D019076:Transgenes; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism*
chemical_list: D000970:Antineoplastic Agents; D016159:Tumor Suppressor Protein p53; D004317:Doxorubicin; D008156:Luciferases
keywords: Adenoassociated virus; Bioluminescence imaging; Doxorubicin; Gene regulation; Gene therapy; Hypoxia; Mechanical stress; Physiologic stress; Viral vector; p53