Development and characterization of an enhanced nonviral expression vector for electroporation cancer treatment
Nonviral plasmid DNA gene therapy represents a promising approach for the treatment of many diseases including cancer. Intracellular delivery of DNA can be achieved with the application of electroporation, which facilitates the initial transport of exogenous DNA across the cell membrane into the cyt...
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2014-01-01
|
| Series: | Molecular Therapy: Methods & Clinical Development |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S232905011630078X |
| _version_ | 1811308620666634240 |
|---|---|
| author | Patrick F Forde Lindsay J Hall Mira Sadadcharam Marcle de Kruijf Gerald C O'Sullivan Declan M Soden |
| author_facet | Patrick F Forde Lindsay J Hall Mira Sadadcharam Marcle de Kruijf Gerald C O'Sullivan Declan M Soden |
| author_sort | Patrick F Forde |
| collection | DOAJ |
| description | Nonviral plasmid DNA gene therapy represents a promising approach for the treatment of many diseases including cancer. Intracellular delivery of DNA can be achieved with the application of electroporation, which facilitates the initial transport of exogenous DNA across the cell membrane into the cytoplasm. However, it does not guarantee further transport of the DNA from the cytoplasm to the nucleus for subsequent mRNA expression, resulting in varying degrees of exogenous gene translation and a major limitation in comparison to viral approaches. To overcome these expression difficulties, we developed a proof-of-concept vector enhanced expression vector (EEV), which incorporates elements from viral systems including nuclear localization sequences and a viral replicase from the Semliki Forest virus. The replicase allows for cytoplasmic mRNA expression and bypasses the need for nuclear localization to generate high levels of gene expression. We have demonstrated that our EEV is capable of achieving high levels of expression in a variety of tissue types. Antitumor effects of pEEV were demonstrated by the delayed growth and increased survival of the nontherapeutic pEEV-treated CT26 tumor model. Using a novel endoscopic electroporation system, EndoVe, we demonstrate and compare, for the first time, both standard cytomegalovirus (CMV) promoter-driven plasmid and EEV gene expression in intraluminal porcine tissues. Our EEV plasmid displays reliable and superior expression capability, and due to its inherent induced oncolytic activity in transfected cells, it may enhance the efficacy and safety of several cancer immunogene therapy approaches. |
| first_indexed | 2024-04-13T09:26:30Z |
| format | Article |
| id | doaj.art-41ba104c43a147a0a16e7e102d481248 |
| institution | Directory Open Access Journal |
| issn | 2329-0501 |
| language | English |
| last_indexed | 2024-04-13T09:26:30Z |
| publishDate | 2014-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Therapy: Methods & Clinical Development |
| spelling | doaj.art-41ba104c43a147a0a16e7e102d4812482022-12-22T02:52:25ZengElsevierMolecular Therapy: Methods & Clinical Development2329-05012014-01-011C10.1038/mtm.2014.12Development and characterization of an enhanced nonviral expression vector for electroporation cancer treatmentPatrick F Forde0Lindsay J Hall1Mira Sadadcharam2Marcle de Kruijf3Gerald C O'Sullivan4Declan M Soden5Cork Cancer Research Centre, Leslie C Quick Laboratory, BioSciences Institute, University College Cork, Cork, IrelandNorwich Medical School, University of East Anglia, Norwich, UKCork Cancer Research Centre, Leslie C Quick Laboratory, BioSciences Institute, University College Cork, Cork, IrelandCork Cancer Research Centre, Leslie C Quick Laboratory, BioSciences Institute, University College Cork, Cork, IrelandCork Cancer Research Centre, Leslie C Quick Laboratory, BioSciences Institute, University College Cork, Cork, IrelandCork Cancer Research Centre, Leslie C Quick Laboratory, BioSciences Institute, University College Cork, Cork, IrelandNonviral plasmid DNA gene therapy represents a promising approach for the treatment of many diseases including cancer. Intracellular delivery of DNA can be achieved with the application of electroporation, which facilitates the initial transport of exogenous DNA across the cell membrane into the cytoplasm. However, it does not guarantee further transport of the DNA from the cytoplasm to the nucleus for subsequent mRNA expression, resulting in varying degrees of exogenous gene translation and a major limitation in comparison to viral approaches. To overcome these expression difficulties, we developed a proof-of-concept vector enhanced expression vector (EEV), which incorporates elements from viral systems including nuclear localization sequences and a viral replicase from the Semliki Forest virus. The replicase allows for cytoplasmic mRNA expression and bypasses the need for nuclear localization to generate high levels of gene expression. We have demonstrated that our EEV is capable of achieving high levels of expression in a variety of tissue types. Antitumor effects of pEEV were demonstrated by the delayed growth and increased survival of the nontherapeutic pEEV-treated CT26 tumor model. Using a novel endoscopic electroporation system, EndoVe, we demonstrate and compare, for the first time, both standard cytomegalovirus (CMV) promoter-driven plasmid and EEV gene expression in intraluminal porcine tissues. Our EEV plasmid displays reliable and superior expression capability, and due to its inherent induced oncolytic activity in transfected cells, it may enhance the efficacy and safety of several cancer immunogene therapy approaches.http://www.sciencedirect.com/science/article/pii/S232905011630078X |
| spellingShingle | Patrick F Forde Lindsay J Hall Mira Sadadcharam Marcle de Kruijf Gerald C O'Sullivan Declan M Soden Development and characterization of an enhanced nonviral expression vector for electroporation cancer treatment Molecular Therapy: Methods & Clinical Development |
| title | Development and characterization of an enhanced nonviral expression vector for electroporation cancer treatment |
| title_full | Development and characterization of an enhanced nonviral expression vector for electroporation cancer treatment |
| title_fullStr | Development and characterization of an enhanced nonviral expression vector for electroporation cancer treatment |
| title_full_unstemmed | Development and characterization of an enhanced nonviral expression vector for electroporation cancer treatment |
| title_short | Development and characterization of an enhanced nonviral expression vector for electroporation cancer treatment |
| title_sort | development and characterization of an enhanced nonviral expression vector for electroporation cancer treatment |
| url | http://www.sciencedirect.com/science/article/pii/S232905011630078X |
| work_keys_str_mv | AT patrickfforde developmentandcharacterizationofanenhancednonviralexpressionvectorforelectroporationcancertreatment AT lindsayjhall developmentandcharacterizationofanenhancednonviralexpressionvectorforelectroporationcancertreatment AT mirasadadcharam developmentandcharacterizationofanenhancednonviralexpressionvectorforelectroporationcancertreatment AT marcledekruijf developmentandcharacterizationofanenhancednonviralexpressionvectorforelectroporationcancertreatment AT geraldcosullivan developmentandcharacterizationofanenhancednonviralexpressionvectorforelectroporationcancertreatment AT declanmsoden developmentandcharacterizationofanenhancednonviralexpressionvectorforelectroporationcancertreatment |