Minicircle DNA Provides Enhanced and Prolonged Transgene Expression Following Airway Gene Transfer

Gene therapy for cystic fibrosis using non-viral, plasmid-based formulations has been the subject of intensive research for over two decades but a clinically viable product has yet to materialise in large part due to inefficient transgene expression. Minicircle DNA give enhanced and more persistent...

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Main Authors:	Munye, Mustafa M., Tagalakis, Aristides D., Barnes, Josephine L., Brown, Rachel E., McAnulty, Robin J., Howe, Steven J., Hart, Stephen L.
Format:	Online
Language:	English
Published:	Nature Publishing Group 2016
Online Access:	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4792149/

id	pubmed-4792149
recordtype	oai_dc
spelling	pubmed-47921492016-03-16 Minicircle DNA Provides Enhanced and Prolonged Transgene Expression Following Airway Gene Transfer Munye, Mustafa M. Tagalakis, Aristides D. Barnes, Josephine L. Brown, Rachel E. McAnulty, Robin J. Howe, Steven J. Hart, Stephen L. Article Gene therapy for cystic fibrosis using non-viral, plasmid-based formulations has been the subject of intensive research for over two decades but a clinically viable product has yet to materialise in large part due to inefficient transgene expression. Minicircle DNA give enhanced and more persistent transgene expression compared to plasmid DNA in a number of organ systems but has not been assessed in the lung. In this study we compared minicircle DNA with plasmid DNA in transfections of airway epithelial cells. In vitro, luciferase gene expression from minicircles was 5–10-fold higher than with plasmid DNA. In eGFP transfections in vitro both the mean fluorescence intensity and percentage of cells transfected was 2–4-fold higher with minicircle DNA. Administration of equimolar amounts of DNA to mouse lungs resulted in a reduced inflammatory response and more persistent transgene expression, with luciferase activity persisting for 2 weeks from minicircle DNA compared to plasmid formulations. Transfection of equal mass amounts of DNA in mouse lungs resulted in a 6-fold increase in transgene expression in addition to more persistent transgene expression. Our findings have clear implications for gene therapy of airway disorders where plasmid DNA transfections have so far proven inefficient in clinical trials. Nature Publishing Group 2016-03-15 /pmc/articles/PMC4792149/ /pubmed/26975732 http://dx.doi.org/10.1038/srep23125 Text en Copyright © 2016, Macmillan Publishers Limited http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
repository_type	Open Access Journal
institution_category	Foreign Institution
institution	US National Center for Biotechnology Information
building	NCBI PubMed
collection	Online Access
language	English
format	Online
author	Munye, Mustafa M. Tagalakis, Aristides D. Barnes, Josephine L. Brown, Rachel E. McAnulty, Robin J. Howe, Steven J. Hart, Stephen L.
spellingShingle	Munye, Mustafa M. Tagalakis, Aristides D. Barnes, Josephine L. Brown, Rachel E. McAnulty, Robin J. Howe, Steven J. Hart, Stephen L. Minicircle DNA Provides Enhanced and Prolonged Transgene Expression Following Airway Gene Transfer
author_facet	Munye, Mustafa M. Tagalakis, Aristides D. Barnes, Josephine L. Brown, Rachel E. McAnulty, Robin J. Howe, Steven J. Hart, Stephen L.
author_sort	Munye, Mustafa M.
title	Minicircle DNA Provides Enhanced and Prolonged Transgene Expression Following Airway Gene Transfer
title_short	Minicircle DNA Provides Enhanced and Prolonged Transgene Expression Following Airway Gene Transfer
title_full	Minicircle DNA Provides Enhanced and Prolonged Transgene Expression Following Airway Gene Transfer
title_fullStr	Minicircle DNA Provides Enhanced and Prolonged Transgene Expression Following Airway Gene Transfer
title_full_unstemmed	Minicircle DNA Provides Enhanced and Prolonged Transgene Expression Following Airway Gene Transfer
title_sort	minicircle dna provides enhanced and prolonged transgene expression following airway gene transfer
description	Gene therapy for cystic fibrosis using non-viral, plasmid-based formulations has been the subject of intensive research for over two decades but a clinically viable product has yet to materialise in large part due to inefficient transgene expression. Minicircle DNA give enhanced and more persistent transgene expression compared to plasmid DNA in a number of organ systems but has not been assessed in the lung. In this study we compared minicircle DNA with plasmid DNA in transfections of airway epithelial cells. In vitro, luciferase gene expression from minicircles was 5–10-fold higher than with plasmid DNA. In eGFP transfections in vitro both the mean fluorescence intensity and percentage of cells transfected was 2–4-fold higher with minicircle DNA. Administration of equimolar amounts of DNA to mouse lungs resulted in a reduced inflammatory response and more persistent transgene expression, with luciferase activity persisting for 2 weeks from minicircle DNA compared to plasmid formulations. Transfection of equal mass amounts of DNA in mouse lungs resulted in a 6-fold increase in transgene expression in addition to more persistent transgene expression. Our findings have clear implications for gene therapy of airway disorders where plasmid DNA transfections have so far proven inefficient in clinical trials.
publisher	Nature Publishing Group
publishDate	2016
url	https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4792149/
_version_	1613551961081118720

Minicircle DNA Provides Enhanced and Prolonged Transgene Expression Following Airway Gene Transfer

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