Cargando…
Exosome-associated SUMOylation mutant AAV demonstrates improved ocular gene transfer efficiency in vivo
Exosome associated Adeno-associated virus (AAV) vectors have emerged as a promising tool in gene therapy. Recently, we elucidated the role of SUMOylation post-translational modification in AAV2 capsid and demonstrated that capsid modifications at SUMOylation sites, enhance vector transduction. The p...
Autores principales: | , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
2020
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7212041/ https://www.ncbi.nlm.nih.gov/pubmed/32302639 http://dx.doi.org/10.1016/j.virusres.2020.197966 |
_version_ | 1783531551946964992 |
---|---|
author | Maurya, Shubham Jayandharan, Giridhara R. |
author_facet | Maurya, Shubham Jayandharan, Giridhara R. |
author_sort | Maurya, Shubham |
collection | PubMed |
description | Exosome associated Adeno-associated virus (AAV) vectors have emerged as a promising tool in gene therapy. Recently, we elucidated the role of SUMOylation post-translational modification in AAV2 capsid and demonstrated that capsid modifications at SUMOylation sites, enhance vector transduction. The present study was designed to study the combinatorial effect of exosome delivery of a SUMOylation site modified AAV2, during ocular gene therapy. In the first set of experiments, we investigated the in vitro gene transfer potential of exo-some-associated SUMOylation mutant AAV2 (Exo-K105Q-EGFP) in human retinal pigmental epithelial (ARPE19) cells. Our data showed that, Exo-K105Q vectors had a significantly higher transduction potential in ARPE19 cells when compared to exosomes derived from wildtype AAV2 (Exo-AAV2-EGFP) vector packaging. Subsequently, an intravitreal administration of exosome associated mutant AAV2 vectors in C57BL6/J mice, demonstrated a significant increase reporter gene (EFGP) expression 4 weeks after gene transfer. Further immunostaining, revealed that these exosome-based vectors also had a better permeation across the retinal layers. These data highlight the translational potential of exosome associated SUMOylation mutant AAV for ocular gene therapy. |
format | Online Article Text |
id | pubmed-7212041 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
record_format | MEDLINE/PubMed |
spelling | pubmed-72120412020-05-11 Exosome-associated SUMOylation mutant AAV demonstrates improved ocular gene transfer efficiency in vivo Maurya, Shubham Jayandharan, Giridhara R. Virus Res Article Exosome associated Adeno-associated virus (AAV) vectors have emerged as a promising tool in gene therapy. Recently, we elucidated the role of SUMOylation post-translational modification in AAV2 capsid and demonstrated that capsid modifications at SUMOylation sites, enhance vector transduction. The present study was designed to study the combinatorial effect of exosome delivery of a SUMOylation site modified AAV2, during ocular gene therapy. In the first set of experiments, we investigated the in vitro gene transfer potential of exo-some-associated SUMOylation mutant AAV2 (Exo-K105Q-EGFP) in human retinal pigmental epithelial (ARPE19) cells. Our data showed that, Exo-K105Q vectors had a significantly higher transduction potential in ARPE19 cells when compared to exosomes derived from wildtype AAV2 (Exo-AAV2-EGFP) vector packaging. Subsequently, an intravitreal administration of exosome associated mutant AAV2 vectors in C57BL6/J mice, demonstrated a significant increase reporter gene (EFGP) expression 4 weeks after gene transfer. Further immunostaining, revealed that these exosome-based vectors also had a better permeation across the retinal layers. These data highlight the translational potential of exosome associated SUMOylation mutant AAV for ocular gene therapy. 2020-04-14 2020-04-14 /pmc/articles/PMC7212041/ /pubmed/32302639 http://dx.doi.org/10.1016/j.virusres.2020.197966 Text en http://creativecommons.org/licenses/BY-NC-ND/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/BY-NC-ND/4.0/). |
spellingShingle | Article Maurya, Shubham Jayandharan, Giridhara R. Exosome-associated SUMOylation mutant AAV demonstrates improved ocular gene transfer efficiency in vivo |
title | Exosome-associated SUMOylation mutant AAV demonstrates improved ocular gene transfer efficiency in vivo |
title_full | Exosome-associated SUMOylation mutant AAV demonstrates improved ocular gene transfer efficiency in vivo |
title_fullStr | Exosome-associated SUMOylation mutant AAV demonstrates improved ocular gene transfer efficiency in vivo |
title_full_unstemmed | Exosome-associated SUMOylation mutant AAV demonstrates improved ocular gene transfer efficiency in vivo |
title_short | Exosome-associated SUMOylation mutant AAV demonstrates improved ocular gene transfer efficiency in vivo |
title_sort | exosome-associated sumoylation mutant aav demonstrates improved ocular gene transfer efficiency in vivo |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7212041/ https://www.ncbi.nlm.nih.gov/pubmed/32302639 http://dx.doi.org/10.1016/j.virusres.2020.197966 |
work_keys_str_mv | AT mauryashubham exosomeassociatedsumoylationmutantaavdemonstratesimprovedoculargenetransferefficiencyinvivo AT jayandharangiridharar exosomeassociatedsumoylationmutantaavdemonstratesimprovedoculargenetransferefficiencyinvivo |