Cargando…
Transcellular Targeting of Fiber- and Hexon-Modified Adenovirus Vectors across the Brain Microvascular Endothelial Cells In Vitro
In central nervous system (CNS)-directed gene therapy, efficient targeting of brain parenchyma through the vascular route is prevented by the endothelium and the epithelium of the blood-brain and the blood-cerebrospinal fluid barriers, respectively. In this study, we evaluated the feasibility of the...
Autores principales: | , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2012
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3459940/ https://www.ncbi.nlm.nih.gov/pubmed/23029348 http://dx.doi.org/10.1371/journal.pone.0045977 |
_version_ | 1782244888513871872 |
---|---|
author | Laakkonen, Johanna P. Engler, Tatjana Romero, Ignacio A. Weksler, Babette Couraud, Pierre-Olivier Kreppel, Florian Kochanek, Stefan |
author_facet | Laakkonen, Johanna P. Engler, Tatjana Romero, Ignacio A. Weksler, Babette Couraud, Pierre-Olivier Kreppel, Florian Kochanek, Stefan |
author_sort | Laakkonen, Johanna P. |
collection | PubMed |
description | In central nervous system (CNS)-directed gene therapy, efficient targeting of brain parenchyma through the vascular route is prevented by the endothelium and the epithelium of the blood-brain and the blood-cerebrospinal fluid barriers, respectively. In this study, we evaluated the feasibility of the combined genetic and chemical adenovirus capsid modification technology to enable transcellular delivery of targeted adenovirus (Ad) vectors across the blood-brain barrier (BBB) in vitro models. As a proof-of-principle ligand, maleimide-activated full-length human transferrin (hTf) was covalently attached to cysteine-modified Ad serotype 5 vectors either to its fiber or hexon protein. In transcytosis experiments, hTf-coupled vectors were shown to be redirected across the BBB models, the transcytosis activity of the vectors being dependent on the location of the capsid modification and the in vitro model used. The transduction efficiency of hTf-targeted vectors decreased significantly in confluent, polarized cells, indicating that the intracellular route of the vectors differed between unpolarized and polarized cells. After transcellular delivery the majority of the hTf-modified vectors remained intact and partly capable of gene transfer. Altogether, our results demonstrate that i) covalent attachment of a ligand to Ad capsid can mediate transcellular targeting across the cerebral endothelium in vitro, ii) the attachment site of the ligand influences its transcytosis efficiency and iii) combined genetic/chemical modification of Ad vector can be used as a versatile platform for the development of Ad vectors for transcellular targeting. |
format | Online Article Text |
id | pubmed-3459940 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-34599402012-10-01 Transcellular Targeting of Fiber- and Hexon-Modified Adenovirus Vectors across the Brain Microvascular Endothelial Cells In Vitro Laakkonen, Johanna P. Engler, Tatjana Romero, Ignacio A. Weksler, Babette Couraud, Pierre-Olivier Kreppel, Florian Kochanek, Stefan PLoS One Research Article In central nervous system (CNS)-directed gene therapy, efficient targeting of brain parenchyma through the vascular route is prevented by the endothelium and the epithelium of the blood-brain and the blood-cerebrospinal fluid barriers, respectively. In this study, we evaluated the feasibility of the combined genetic and chemical adenovirus capsid modification technology to enable transcellular delivery of targeted adenovirus (Ad) vectors across the blood-brain barrier (BBB) in vitro models. As a proof-of-principle ligand, maleimide-activated full-length human transferrin (hTf) was covalently attached to cysteine-modified Ad serotype 5 vectors either to its fiber or hexon protein. In transcytosis experiments, hTf-coupled vectors were shown to be redirected across the BBB models, the transcytosis activity of the vectors being dependent on the location of the capsid modification and the in vitro model used. The transduction efficiency of hTf-targeted vectors decreased significantly in confluent, polarized cells, indicating that the intracellular route of the vectors differed between unpolarized and polarized cells. After transcellular delivery the majority of the hTf-modified vectors remained intact and partly capable of gene transfer. Altogether, our results demonstrate that i) covalent attachment of a ligand to Ad capsid can mediate transcellular targeting across the cerebral endothelium in vitro, ii) the attachment site of the ligand influences its transcytosis efficiency and iii) combined genetic/chemical modification of Ad vector can be used as a versatile platform for the development of Ad vectors for transcellular targeting. Public Library of Science 2012-09-27 /pmc/articles/PMC3459940/ /pubmed/23029348 http://dx.doi.org/10.1371/journal.pone.0045977 Text en © 2012 Laakkonen 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, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Laakkonen, Johanna P. Engler, Tatjana Romero, Ignacio A. Weksler, Babette Couraud, Pierre-Olivier Kreppel, Florian Kochanek, Stefan Transcellular Targeting of Fiber- and Hexon-Modified Adenovirus Vectors across the Brain Microvascular Endothelial Cells In Vitro |
title | Transcellular Targeting of Fiber- and Hexon-Modified Adenovirus Vectors across the Brain Microvascular Endothelial Cells In Vitro
|
title_full | Transcellular Targeting of Fiber- and Hexon-Modified Adenovirus Vectors across the Brain Microvascular Endothelial Cells In Vitro
|
title_fullStr | Transcellular Targeting of Fiber- and Hexon-Modified Adenovirus Vectors across the Brain Microvascular Endothelial Cells In Vitro
|
title_full_unstemmed | Transcellular Targeting of Fiber- and Hexon-Modified Adenovirus Vectors across the Brain Microvascular Endothelial Cells In Vitro
|
title_short | Transcellular Targeting of Fiber- and Hexon-Modified Adenovirus Vectors across the Brain Microvascular Endothelial Cells In Vitro
|
title_sort | transcellular targeting of fiber- and hexon-modified adenovirus vectors across the brain microvascular endothelial cells in vitro |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3459940/ https://www.ncbi.nlm.nih.gov/pubmed/23029348 http://dx.doi.org/10.1371/journal.pone.0045977 |
work_keys_str_mv | AT laakkonenjohannap transcellulartargetingoffiberandhexonmodifiedadenovirusvectorsacrossthebrainmicrovascularendothelialcellsinvitro AT englertatjana transcellulartargetingoffiberandhexonmodifiedadenovirusvectorsacrossthebrainmicrovascularendothelialcellsinvitro AT romeroignacioa transcellulartargetingoffiberandhexonmodifiedadenovirusvectorsacrossthebrainmicrovascularendothelialcellsinvitro AT wekslerbabette transcellulartargetingoffiberandhexonmodifiedadenovirusvectorsacrossthebrainmicrovascularendothelialcellsinvitro AT couraudpierreolivier transcellulartargetingoffiberandhexonmodifiedadenovirusvectorsacrossthebrainmicrovascularendothelialcellsinvitro AT kreppelflorian transcellulartargetingoffiberandhexonmodifiedadenovirusvectorsacrossthebrainmicrovascularendothelialcellsinvitro AT kochanekstefan transcellulartargetingoffiberandhexonmodifiedadenovirusvectorsacrossthebrainmicrovascularendothelialcellsinvitro |