Multigenic lentiviral vectors for combined and tissue-specific expression of miRNA- and protein-based antiangiogenic factors
Lentivirus-based gene delivery vectors carrying multiple gene cassettes are powerful tools in gene transfer studies and gene therapy, allowing coexpression of multiple therapeutic factors and, if desired, fluorescent reporters. Current strategies to express transgenes and microRNA (miRNA) clusters f...
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Series: | Molecular Therapy: Methods & Clinical Development |
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doaj-22d8b8cf60154d25b2420e8d2fe941732020-11-25T02:06:25ZengElsevierMolecular Therapy: Methods & Clinical Development2329-05012015-01-012Multigenic lentiviral vectors for combined and tissue-specific expression of miRNA- and protein-based antiangiogenic factorsAnne Louise Askou0Lars Aagaard1Corinne Kostic2Yvan Arsenijevic3Anne Kruse Hollensen4Toke Bek5Thomas Gryesten Jensen6Jacob Giehm Mikkelsen7Thomas Juhl Corydon8Department of Biomedicine, Aarhus University, Aarhus, DenmarkDepartment of Biomedicine, Aarhus University, Aarhus, DenmarkDepartment of Ophthalmology, Unit of Gene Therapy and Stem Cell Biology, University of Lausanne, Jules-Gonin Eye Hospital, Lausanne, SwitzerlandDepartment of Ophthalmology, Unit of Gene Therapy and Stem Cell Biology, University of Lausanne, Jules-Gonin Eye Hospital, Lausanne, SwitzerlandDepartment of Biomedicine, Aarhus University, Aarhus, DenmarkDepartment of Ophthalmology, Aarhus University Hospital, Aarhus, DenmarkDepartment of Biomedicine, Aarhus University, Aarhus, DenmarkDepartment of Biomedicine, Aarhus University, Aarhus, DenmarkDepartment of Biomedicine, Aarhus University, Aarhus, DenmarkLentivirus-based gene delivery vectors carrying multiple gene cassettes are powerful tools in gene transfer studies and gene therapy, allowing coexpression of multiple therapeutic factors and, if desired, fluorescent reporters. Current strategies to express transgenes and microRNA (miRNA) clusters from a single vector have certain limitations that affect transgene expression levels and/or vector titers. In this study, we describe a novel vector design that facilitates combined expression of therapeutic RNA- and protein-based antiangiogenic factors as well as a fluorescent reporter from back-to-back RNApolII-driven expression cassettes. This configuration allows effective production of intron-embedded miRNAs that are released upon transduction of target cells. Exploiting such multigenic lentiviral vectors, we demonstrate robust miRNA-directed downregulation of vascular endothelial growth factor (VEGF) expression, leading to reduced angiogenesis, and parallel impairment of angiogenic pathways by codelivering the gene encoding pigment epithelium-derived factor (PEDF). Notably, subretinal injections of lentiviral vectors reveal efficient retinal pigment epithelium-specific gene expression driven by the VMD2 promoter, verifying that multigenic lentiviral vectors can be produced with high titers sufficient for in vivo applications. Altogether, our results suggest the potential applicability of combined miRNA- and protein-encoding lentiviral vectors in antiangiogenic gene therapy, including new combination therapies for amelioration of age-related macular degeneration.http://www.sciencedirect.com/science/article/pii/S2329050116300080 |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Anne Louise Askou Lars Aagaard Corinne Kostic Yvan Arsenijevic Anne Kruse Hollensen Toke Bek Thomas Gryesten Jensen Jacob Giehm Mikkelsen Thomas Juhl Corydon |
spellingShingle |
Anne Louise Askou Lars Aagaard Corinne Kostic Yvan Arsenijevic Anne Kruse Hollensen Toke Bek Thomas Gryesten Jensen Jacob Giehm Mikkelsen Thomas Juhl Corydon Multigenic lentiviral vectors for combined and tissue-specific expression of miRNA- and protein-based antiangiogenic factors Molecular Therapy: Methods & Clinical Development |
author_facet |
Anne Louise Askou Lars Aagaard Corinne Kostic Yvan Arsenijevic Anne Kruse Hollensen Toke Bek Thomas Gryesten Jensen Jacob Giehm Mikkelsen Thomas Juhl Corydon |
author_sort |
Anne Louise Askou |
title |
Multigenic lentiviral vectors for combined and tissue-specific expression of miRNA- and protein-based antiangiogenic factors |
title_short |
Multigenic lentiviral vectors for combined and tissue-specific expression of miRNA- and protein-based antiangiogenic factors |
title_full |
Multigenic lentiviral vectors for combined and tissue-specific expression of miRNA- and protein-based antiangiogenic factors |
title_fullStr |
Multigenic lentiviral vectors for combined and tissue-specific expression of miRNA- and protein-based antiangiogenic factors |
title_full_unstemmed |
Multigenic lentiviral vectors for combined and tissue-specific expression of miRNA- and protein-based antiangiogenic factors |
title_sort |
multigenic lentiviral vectors for combined and tissue-specific expression of mirna- and protein-based antiangiogenic factors |
publisher |
Elsevier |
series |
Molecular Therapy: Methods & Clinical Development |
issn |
2329-0501 |
publishDate |
2015-01-01 |
description |
Lentivirus-based gene delivery vectors carrying multiple gene cassettes are powerful tools in gene transfer studies and gene therapy, allowing coexpression of multiple therapeutic factors and, if desired, fluorescent reporters. Current strategies to express transgenes and microRNA (miRNA) clusters from a single vector have certain limitations that affect transgene expression levels and/or vector titers. In this study, we describe a novel vector design that facilitates combined expression of therapeutic RNA- and protein-based antiangiogenic factors as well as a fluorescent reporter from back-to-back RNApolII-driven expression cassettes. This configuration allows effective production of intron-embedded miRNAs that are released upon transduction of target cells. Exploiting such multigenic lentiviral vectors, we demonstrate robust miRNA-directed downregulation of vascular endothelial growth factor (VEGF) expression, leading to reduced angiogenesis, and parallel impairment of angiogenic pathways by codelivering the gene encoding pigment epithelium-derived factor (PEDF). Notably, subretinal injections of lentiviral vectors reveal efficient retinal pigment epithelium-specific gene expression driven by the VMD2 promoter, verifying that multigenic lentiviral vectors can be produced with high titers sufficient for in vivo applications. Altogether, our results suggest the potential applicability of combined miRNA- and protein-encoding lentiviral vectors in antiangiogenic gene therapy, including new combination therapies for amelioration of age-related macular degeneration. |
url |
http://www.sciencedirect.com/science/article/pii/S2329050116300080 |
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