Cargando…

Studying Closed Hydrodynamic Models of “In Vivo” DNA Perfusion in Pig Liver for Gene Therapy Translation to Humans

INTRODUCTION: Expressing exogenous genes after naked DNA delivery into hepatocytes might achieve sustained and high expression of human proteins. Tail vein DNA injection is an efficient procedure for gene transfer in murine liver. Hydrodynamic procedures in large animals require organ targeting, and...

Descripción completa

Detalles Bibliográficos
Autores principales: Sendra, Luis, Miguel, Antonio, Pérez-Enguix, Daniel, Herrero, María José, Montalvá, Eva, García-Gimeno, María Adelaida, Noguera, Inmaculada, Díaz, Ana, Pérez, Judith, Sanz, Pascual, López-Andújar, Rafael, Martí-Bonmatí, Luis, Aliño, Salvador F.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5047531/
https://www.ncbi.nlm.nih.gov/pubmed/27695064
http://dx.doi.org/10.1371/journal.pone.0163898
_version_ 1782457431728586752
author Sendra, Luis
Miguel, Antonio
Pérez-Enguix, Daniel
Herrero, María José
Montalvá, Eva
García-Gimeno, María Adelaida
Noguera, Inmaculada
Díaz, Ana
Pérez, Judith
Sanz, Pascual
López-Andújar, Rafael
Martí-Bonmatí, Luis
Aliño, Salvador F.
author_facet Sendra, Luis
Miguel, Antonio
Pérez-Enguix, Daniel
Herrero, María José
Montalvá, Eva
García-Gimeno, María Adelaida
Noguera, Inmaculada
Díaz, Ana
Pérez, Judith
Sanz, Pascual
López-Andújar, Rafael
Martí-Bonmatí, Luis
Aliño, Salvador F.
author_sort Sendra, Luis
collection PubMed
description INTRODUCTION: Expressing exogenous genes after naked DNA delivery into hepatocytes might achieve sustained and high expression of human proteins. Tail vein DNA injection is an efficient procedure for gene transfer in murine liver. Hydrodynamic procedures in large animals require organ targeting, and improve with liver vascular exclusion. In the present study, two closed liver hydrofection models employing the human alpha-1-antitrypsin (hAAT) gene are compared to reference standards in order to evaluate their potential clinical interest. MATERIAL AND METHODS: A solution of naked DNA bearing the hAAT gene was retrogradely injected in 7 pig livers using two different closed perfusion procedures: an endovascular catheterization-mediated procedure (n = 3) with infrahepatic inferior vena cava and portal vein blockage; and a surgery-mediated procedure (n = 4) with completely sealed liver. Gene transfer was performed through the suprahepatic inferior cava vein in the endovascular procedure and through the infrahepatic inferior vena cava in the surgical procedure. The efficiency of the procedures was evaluated 14 days after hydrofection by quantifying the hAAT protein copies per cell in tissue and in plasma. For comparison, samples from mice (n = 7) successfully hydrofected with hAAT and healthy human liver segments (n = 4) were evaluated. RESULTS: Gene decoding occurs efficiently using both procedures, with liver vascular arrest improving its efficiency. The surgically closed procedure (sealed organ) reached higher tissue protein levels (4x10^5- copies/cell) than the endovascular procedure, though the levels were lower than in human liver (5x10^6- copies/cell) and hydrofected mouse liver (10^6- copies/cell). However, protein levels in plasma were lower (p<0.001) than the reference standards in all cases. CONCLUSION: Hydrofection of hAAT DNA to “in vivo” isolated pig liver mediates highly efficient gene delivery and protein expression in tissue. Both endovascular and surgically closed models mediate high tissue protein expression. Impairment of protein secretion to plasma is observed and might be species-related. This study reinforces the potential application of closed liver hydrofection for therapeutic purposes, provided protein secretion improves.
format Online
Article
Text
id pubmed-5047531
institution National Center for Biotechnology Information
language English
publishDate 2016
publisher Public Library of Science
record_format MEDLINE/PubMed
spelling pubmed-50475312016-10-27 Studying Closed Hydrodynamic Models of “In Vivo” DNA Perfusion in Pig Liver for Gene Therapy Translation to Humans Sendra, Luis Miguel, Antonio Pérez-Enguix, Daniel Herrero, María José Montalvá, Eva García-Gimeno, María Adelaida Noguera, Inmaculada Díaz, Ana Pérez, Judith Sanz, Pascual López-Andújar, Rafael Martí-Bonmatí, Luis Aliño, Salvador F. PLoS One Research Article INTRODUCTION: Expressing exogenous genes after naked DNA delivery into hepatocytes might achieve sustained and high expression of human proteins. Tail vein DNA injection is an efficient procedure for gene transfer in murine liver. Hydrodynamic procedures in large animals require organ targeting, and improve with liver vascular exclusion. In the present study, two closed liver hydrofection models employing the human alpha-1-antitrypsin (hAAT) gene are compared to reference standards in order to evaluate their potential clinical interest. MATERIAL AND METHODS: A solution of naked DNA bearing the hAAT gene was retrogradely injected in 7 pig livers using two different closed perfusion procedures: an endovascular catheterization-mediated procedure (n = 3) with infrahepatic inferior vena cava and portal vein blockage; and a surgery-mediated procedure (n = 4) with completely sealed liver. Gene transfer was performed through the suprahepatic inferior cava vein in the endovascular procedure and through the infrahepatic inferior vena cava in the surgical procedure. The efficiency of the procedures was evaluated 14 days after hydrofection by quantifying the hAAT protein copies per cell in tissue and in plasma. For comparison, samples from mice (n = 7) successfully hydrofected with hAAT and healthy human liver segments (n = 4) were evaluated. RESULTS: Gene decoding occurs efficiently using both procedures, with liver vascular arrest improving its efficiency. The surgically closed procedure (sealed organ) reached higher tissue protein levels (4x10^5- copies/cell) than the endovascular procedure, though the levels were lower than in human liver (5x10^6- copies/cell) and hydrofected mouse liver (10^6- copies/cell). However, protein levels in plasma were lower (p<0.001) than the reference standards in all cases. CONCLUSION: Hydrofection of hAAT DNA to “in vivo” isolated pig liver mediates highly efficient gene delivery and protein expression in tissue. Both endovascular and surgically closed models mediate high tissue protein expression. Impairment of protein secretion to plasma is observed and might be species-related. This study reinforces the potential application of closed liver hydrofection for therapeutic purposes, provided protein secretion improves. Public Library of Science 2016-10-03 /pmc/articles/PMC5047531/ /pubmed/27695064 http://dx.doi.org/10.1371/journal.pone.0163898 Text en © 2016 Sendra et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Sendra, Luis
Miguel, Antonio
Pérez-Enguix, Daniel
Herrero, María José
Montalvá, Eva
García-Gimeno, María Adelaida
Noguera, Inmaculada
Díaz, Ana
Pérez, Judith
Sanz, Pascual
López-Andújar, Rafael
Martí-Bonmatí, Luis
Aliño, Salvador F.
Studying Closed Hydrodynamic Models of “In Vivo” DNA Perfusion in Pig Liver for Gene Therapy Translation to Humans
title Studying Closed Hydrodynamic Models of “In Vivo” DNA Perfusion in Pig Liver for Gene Therapy Translation to Humans
title_full Studying Closed Hydrodynamic Models of “In Vivo” DNA Perfusion in Pig Liver for Gene Therapy Translation to Humans
title_fullStr Studying Closed Hydrodynamic Models of “In Vivo” DNA Perfusion in Pig Liver for Gene Therapy Translation to Humans
title_full_unstemmed Studying Closed Hydrodynamic Models of “In Vivo” DNA Perfusion in Pig Liver for Gene Therapy Translation to Humans
title_short Studying Closed Hydrodynamic Models of “In Vivo” DNA Perfusion in Pig Liver for Gene Therapy Translation to Humans
title_sort studying closed hydrodynamic models of “in vivo” dna perfusion in pig liver for gene therapy translation to humans
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5047531/
https://www.ncbi.nlm.nih.gov/pubmed/27695064
http://dx.doi.org/10.1371/journal.pone.0163898
work_keys_str_mv AT sendraluis studyingclosedhydrodynamicmodelsofinvivodnaperfusioninpigliverforgenetherapytranslationtohumans
AT miguelantonio studyingclosedhydrodynamicmodelsofinvivodnaperfusioninpigliverforgenetherapytranslationtohumans
AT perezenguixdaniel studyingclosedhydrodynamicmodelsofinvivodnaperfusioninpigliverforgenetherapytranslationtohumans
AT herreromariajose studyingclosedhydrodynamicmodelsofinvivodnaperfusioninpigliverforgenetherapytranslationtohumans
AT montalvaeva studyingclosedhydrodynamicmodelsofinvivodnaperfusioninpigliverforgenetherapytranslationtohumans
AT garciagimenomariaadelaida studyingclosedhydrodynamicmodelsofinvivodnaperfusioninpigliverforgenetherapytranslationtohumans
AT noguerainmaculada studyingclosedhydrodynamicmodelsofinvivodnaperfusioninpigliverforgenetherapytranslationtohumans
AT diazana studyingclosedhydrodynamicmodelsofinvivodnaperfusioninpigliverforgenetherapytranslationtohumans
AT perezjudith studyingclosedhydrodynamicmodelsofinvivodnaperfusioninpigliverforgenetherapytranslationtohumans
AT sanzpascual studyingclosedhydrodynamicmodelsofinvivodnaperfusioninpigliverforgenetherapytranslationtohumans
AT lopezandujarrafael studyingclosedhydrodynamicmodelsofinvivodnaperfusioninpigliverforgenetherapytranslationtohumans
AT martibonmatiluis studyingclosedhydrodynamicmodelsofinvivodnaperfusioninpigliverforgenetherapytranslationtohumans
AT alinosalvadorf studyingclosedhydrodynamicmodelsofinvivodnaperfusioninpigliverforgenetherapytranslationtohumans