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...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Johanna P Laakkonen, Tatjana Engler, Ignacio A Romero, Babette Weksler, Pierre-Olivier Couraud, Florian Kreppel, Stefan Kochanek
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2012
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/8d9d78060e9e41d6a35dc4201a2691e9
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:8d9d78060e9e41d6a35dc4201a2691e9
record_format	dspace
spelling	oai:doaj.org-article:8d9d78060e9e41d6a35dc4201a2691e92021-11-18T07:04:10ZTranscellular targeting of fiber- and hexon-modified adenovirus vectors across the brain microvascular endothelial cells in vitro.1932-620310.1371/journal.pone.0045977https://doaj.org/article/8d9d78060e9e41d6a35dc4201a2691e92012-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23029348/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203In 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.Johanna P LaakkonenTatjana EnglerIgnacio A RomeroBabette WekslerPierre-Olivier CouraudFlorian KreppelStefan KochanekPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 7, Iss 9, p e45977 (2012)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Johanna P Laakkonen Tatjana Engler Ignacio A Romero Babette Weksler Pierre-Olivier Couraud Florian Kreppel Stefan Kochanek Transcellular targeting of fiber- and hexon-modified adenovirus vectors across the brain microvascular endothelial cells in vitro.
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	article
author	Johanna P Laakkonen Tatjana Engler Ignacio A Romero Babette Weksler Pierre-Olivier Couraud Florian Kreppel Stefan Kochanek
author_facet	Johanna P Laakkonen Tatjana Engler Ignacio A Romero Babette Weksler Pierre-Olivier Couraud Florian Kreppel Stefan Kochanek
author_sort	Johanna P Laakkonen
title	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_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_sort	transcellular targeting of fiber- and hexon-modified adenovirus vectors across the brain microvascular endothelial cells in vitro.
publisher	Public Library of Science (PLoS)
publishDate	2012
url	https://doaj.org/article/8d9d78060e9e41d6a35dc4201a2691e9
work_keys_str_mv	AT johannaplaakkonen transcellulartargetingoffiberandhexonmodifiedadenovirusvectorsacrossthebrainmicrovascularendothelialcellsinvitro AT tatjanaengler transcellulartargetingoffiberandhexonmodifiedadenovirusvectorsacrossthebrainmicrovascularendothelialcellsinvitro AT ignacioaromero transcellulartargetingoffiberandhexonmodifiedadenovirusvectorsacrossthebrainmicrovascularendothelialcellsinvitro AT babetteweksler transcellulartargetingoffiberandhexonmodifiedadenovirusvectorsacrossthebrainmicrovascularendothelialcellsinvitro AT pierreoliviercouraud transcellulartargetingoffiberandhexonmodifiedadenovirusvectorsacrossthebrainmicrovascularendothelialcellsinvitro AT floriankreppel transcellulartargetingoffiberandhexonmodifiedadenovirusvectorsacrossthebrainmicrovascularendothelialcellsinvitro AT stefankochanek transcellulartargetingoffiberandhexonmodifiedadenovirusvectorsacrossthebrainmicrovascularendothelialcellsinvitro
_version_	1718423991989829632

Transcellular targeting of fiber- and hexon-modified adenovirus vectors across the brain microvascular endothelial cells in vitro.

Ejemplares similares