Multigenic lentiviral vectors for combined and tissue-specific expression of miRNA- and protein-based antiangiogenic factors.

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State: Public
Version: Final published version
Serval ID
serval:BIB_4CD7D795B26A
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Multigenic lentiviral vectors for combined and tissue-specific expression of miRNA- and protein-based antiangiogenic factors.
Journal
Molecular Therapy. Methods and Clinical Development
Author(s)
Askou A.L., Aagaard L., Kostic C., Arsenijevic Y., Hollensen A.K., Bek T., Jensen T.G., Mikkelsen J.G., Corydon T.J.
ISSN
2329-0501 (Electronic)
ISSN-L
2329-0501
Publication state
Published
Issued date
2015
Peer-reviewed
Oui
Volume
2
Pages
14064
Language
english
Notes
Publication types: Journal ArticlePublication Status: epublish
Abstract
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.
Pubmed
Open Access
Yes
Create date
25/06/2015 13:32
Last modification date
20/08/2019 14:01
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