A highly efficient, stable, and rapid approach for ex vivo human liver gene therapy via a FLAP lentiviral vector
Autor(a) principal: | |
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Data de Publicação: | 2003 |
Outros Autores: | , , , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | http://hdl.handle.net/10400.4/2187 |
Resumo: | Allogenic hepatocyte transplantation or autologous transplantation of genetically modified hepatocytes has been used successfully to correct congenital or acquired liver diseases and can be considered as an alternative to orthotopic liver transplantation. However, hepatocytes are neither easily maintained in culture nor efficiently genetically modified and are very sensitive to dissociation before their reimplantation into the recipient. These difficulties have greatly limited the use of an ex vivo approach in clinical trials. In the present study, we have shown that primary human and rat hepatocytes can be efficiently transduced with a FLAP lentiviral vector without the need for plating and culture. Efficient transduction of nonadherent primary hepatocytes was achieved with a short period of contact with vector particles, without modifying hepatocyte viability, and using reduced amounts of vector. We also showed that the presence of the DNA FLAP in the vector construct was essential to reach high levels of transduction. Moreover, transplanted into uPA/SCID mouse liver, lentivirally transduced primary human hepatocytes extensively repopulated their liver and maintained a differentiated and functional phenotype as assessed by the stable detection of human albumin and antitrypsin in the serum of the animals for months. In conclusion, the use of FLAP lentiviral vectors allows, in a short period of time, a high transduction efficiency of human functional and reimplantable hepatocytes. This work therefore opens new perspectives for the development of human clinical trials based on liver-directed ex vivo gene therapy. |
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A highly efficient, stable, and rapid approach for ex vivo human liver gene therapy via a FLAP lentiviral vectorTerapia GenéticaVectores GenéticosHepatócitosLentivirus/genéticaRatosAllogenic hepatocyte transplantation or autologous transplantation of genetically modified hepatocytes has been used successfully to correct congenital or acquired liver diseases and can be considered as an alternative to orthotopic liver transplantation. However, hepatocytes are neither easily maintained in culture nor efficiently genetically modified and are very sensitive to dissociation before their reimplantation into the recipient. These difficulties have greatly limited the use of an ex vivo approach in clinical trials. In the present study, we have shown that primary human and rat hepatocytes can be efficiently transduced with a FLAP lentiviral vector without the need for plating and culture. Efficient transduction of nonadherent primary hepatocytes was achieved with a short period of contact with vector particles, without modifying hepatocyte viability, and using reduced amounts of vector. We also showed that the presence of the DNA FLAP in the vector construct was essential to reach high levels of transduction. Moreover, transplanted into uPA/SCID mouse liver, lentivirally transduced primary human hepatocytes extensively repopulated their liver and maintained a differentiated and functional phenotype as assessed by the stable detection of human albumin and antitrypsin in the serum of the animals for months. In conclusion, the use of FLAP lentiviral vectors allows, in a short period of time, a high transduction efficiency of human functional and reimplantable hepatocytes. This work therefore opens new perspectives for the development of human clinical trials based on liver-directed ex vivo gene therapy.RIHUCGiannini, CMorosan, STralhao, JGGuidotti, JEBattaglia, SMollier, KHannoun, LKremsdorf, DGilgenkrantz, HCharneau, P2018-11-28T15:17:05Z2003-072003-07-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10400.4/2187engHepatology. 2003 Jul;38(1):114-22.10.1053/jhep.2003.50265info:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2023-07-11T14:23:31Zoai:rihuc.huc.min-saude.pt:10400.4/2187Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T18:04:39.217385Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse |
dc.title.none.fl_str_mv |
A highly efficient, stable, and rapid approach for ex vivo human liver gene therapy via a FLAP lentiviral vector |
title |
A highly efficient, stable, and rapid approach for ex vivo human liver gene therapy via a FLAP lentiviral vector |
spellingShingle |
A highly efficient, stable, and rapid approach for ex vivo human liver gene therapy via a FLAP lentiviral vector Giannini, C Terapia Genética Vectores Genéticos Hepatócitos Lentivirus/genética Ratos |
title_short |
A highly efficient, stable, and rapid approach for ex vivo human liver gene therapy via a FLAP lentiviral vector |
title_full |
A highly efficient, stable, and rapid approach for ex vivo human liver gene therapy via a FLAP lentiviral vector |
title_fullStr |
A highly efficient, stable, and rapid approach for ex vivo human liver gene therapy via a FLAP lentiviral vector |
title_full_unstemmed |
A highly efficient, stable, and rapid approach for ex vivo human liver gene therapy via a FLAP lentiviral vector |
title_sort |
A highly efficient, stable, and rapid approach for ex vivo human liver gene therapy via a FLAP lentiviral vector |
author |
Giannini, C |
author_facet |
Giannini, C Morosan, S Tralhao, JG Guidotti, JE Battaglia, S Mollier, K Hannoun, L Kremsdorf, D Gilgenkrantz, H Charneau, P |
author_role |
author |
author2 |
Morosan, S Tralhao, JG Guidotti, JE Battaglia, S Mollier, K Hannoun, L Kremsdorf, D Gilgenkrantz, H Charneau, P |
author2_role |
author author author author author author author author author |
dc.contributor.none.fl_str_mv |
RIHUC |
dc.contributor.author.fl_str_mv |
Giannini, C Morosan, S Tralhao, JG Guidotti, JE Battaglia, S Mollier, K Hannoun, L Kremsdorf, D Gilgenkrantz, H Charneau, P |
dc.subject.por.fl_str_mv |
Terapia Genética Vectores Genéticos Hepatócitos Lentivirus/genética Ratos |
topic |
Terapia Genética Vectores Genéticos Hepatócitos Lentivirus/genética Ratos |
description |
Allogenic hepatocyte transplantation or autologous transplantation of genetically modified hepatocytes has been used successfully to correct congenital or acquired liver diseases and can be considered as an alternative to orthotopic liver transplantation. However, hepatocytes are neither easily maintained in culture nor efficiently genetically modified and are very sensitive to dissociation before their reimplantation into the recipient. These difficulties have greatly limited the use of an ex vivo approach in clinical trials. In the present study, we have shown that primary human and rat hepatocytes can be efficiently transduced with a FLAP lentiviral vector without the need for plating and culture. Efficient transduction of nonadherent primary hepatocytes was achieved with a short period of contact with vector particles, without modifying hepatocyte viability, and using reduced amounts of vector. We also showed that the presence of the DNA FLAP in the vector construct was essential to reach high levels of transduction. Moreover, transplanted into uPA/SCID mouse liver, lentivirally transduced primary human hepatocytes extensively repopulated their liver and maintained a differentiated and functional phenotype as assessed by the stable detection of human albumin and antitrypsin in the serum of the animals for months. In conclusion, the use of FLAP lentiviral vectors allows, in a short period of time, a high transduction efficiency of human functional and reimplantable hepatocytes. This work therefore opens new perspectives for the development of human clinical trials based on liver-directed ex vivo gene therapy. |
publishDate |
2003 |
dc.date.none.fl_str_mv |
2003-07 2003-07-01T00:00:00Z 2018-11-28T15:17:05Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://hdl.handle.net/10400.4/2187 |
url |
http://hdl.handle.net/10400.4/2187 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Hepatology. 2003 Jul;38(1):114-22. 10.1053/jhep.2003.50265 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.source.none.fl_str_mv |
reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação instacron:RCAAP |
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Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
instacron_str |
RCAAP |
institution |
RCAAP |
reponame_str |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
collection |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
repository.name.fl_str_mv |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
repository.mail.fl_str_mv |
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1799131708850700288 |