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AAV-Mediated Clarin-1 Expression in the Mouse Retina: Implications for USH3A Gene Therapy

Usher syndrome type III (USH3A) is an autosomal recessive disorder caused by mutations in clarin-1 (CLRN1) gene, leading to progressive retinal degeneration and sensorineural deafness. Efforts to develop therapies for preventing photoreceptor cell loss are hampered by the lack of a retinal phenotype...

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Autores principales: Dinculescu, Astra, Stupay, Rachel M., Deng, Wen-Tao, Dyka, Frank M., Min, Seok-Hong, Boye, Sanford L., Chiodo, Vince A., Abrahan, Carolina E., Zhu, Ping, Li, Qiuhong, Strettoi, Enrica, Novelli, Elena, Nagel-Wolfrum, Kerstin, Wolfrum, Uwe, Smith, W. Clay, Hauswirth, William W.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4755610/
https://www.ncbi.nlm.nih.gov/pubmed/26881841
http://dx.doi.org/10.1371/journal.pone.0148874
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author Dinculescu, Astra
Stupay, Rachel M.
Deng, Wen-Tao
Dyka, Frank M.
Min, Seok-Hong
Boye, Sanford L.
Chiodo, Vince A.
Abrahan, Carolina E.
Zhu, Ping
Li, Qiuhong
Strettoi, Enrica
Novelli, Elena
Nagel-Wolfrum, Kerstin
Wolfrum, Uwe
Smith, W. Clay
Hauswirth, William W.
author_facet Dinculescu, Astra
Stupay, Rachel M.
Deng, Wen-Tao
Dyka, Frank M.
Min, Seok-Hong
Boye, Sanford L.
Chiodo, Vince A.
Abrahan, Carolina E.
Zhu, Ping
Li, Qiuhong
Strettoi, Enrica
Novelli, Elena
Nagel-Wolfrum, Kerstin
Wolfrum, Uwe
Smith, W. Clay
Hauswirth, William W.
author_sort Dinculescu, Astra
collection PubMed
description Usher syndrome type III (USH3A) is an autosomal recessive disorder caused by mutations in clarin-1 (CLRN1) gene, leading to progressive retinal degeneration and sensorineural deafness. Efforts to develop therapies for preventing photoreceptor cell loss are hampered by the lack of a retinal phenotype in the existing USH3 mouse models and by conflicting reports regarding the endogenous retinal localization of clarin-1, a transmembrane protein of unknown function. In this study, we used an AAV-based approach to express CLRN1 in the mouse retina in order to determine the pattern of its subcellular localization in different cell types. We found that all major classes of retinal cells express AAV-delivered CLRN1 driven by the ubiquitous, constitutive small chicken β-actin promoter, which has important implications for the design of future USH3 gene therapy studies. Within photoreceptor cells, AAV-expressed CLRN1 is mainly localized at the inner segment region and outer plexiform layer, similar to the endogenous expression of other usher proteins. Subretinal delivery using a full strength viral titer led to significant loss of retinal function as evidenced by ERG analysis, suggesting that there is a critical limit for CLRN1 expression in photoreceptor cells. Taken together, these results suggest that CLRN1 expression is potentially supported by a variety of retinal cells, and the right combination of AAV vector dose, promoter, and delivery method needs to be selected to develop safe therapies for USH3 disorder.
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spelling pubmed-47556102016-02-26 AAV-Mediated Clarin-1 Expression in the Mouse Retina: Implications for USH3A Gene Therapy Dinculescu, Astra Stupay, Rachel M. Deng, Wen-Tao Dyka, Frank M. Min, Seok-Hong Boye, Sanford L. Chiodo, Vince A. Abrahan, Carolina E. Zhu, Ping Li, Qiuhong Strettoi, Enrica Novelli, Elena Nagel-Wolfrum, Kerstin Wolfrum, Uwe Smith, W. Clay Hauswirth, William W. PLoS One Research Article Usher syndrome type III (USH3A) is an autosomal recessive disorder caused by mutations in clarin-1 (CLRN1) gene, leading to progressive retinal degeneration and sensorineural deafness. Efforts to develop therapies for preventing photoreceptor cell loss are hampered by the lack of a retinal phenotype in the existing USH3 mouse models and by conflicting reports regarding the endogenous retinal localization of clarin-1, a transmembrane protein of unknown function. In this study, we used an AAV-based approach to express CLRN1 in the mouse retina in order to determine the pattern of its subcellular localization in different cell types. We found that all major classes of retinal cells express AAV-delivered CLRN1 driven by the ubiquitous, constitutive small chicken β-actin promoter, which has important implications for the design of future USH3 gene therapy studies. Within photoreceptor cells, AAV-expressed CLRN1 is mainly localized at the inner segment region and outer plexiform layer, similar to the endogenous expression of other usher proteins. Subretinal delivery using a full strength viral titer led to significant loss of retinal function as evidenced by ERG analysis, suggesting that there is a critical limit for CLRN1 expression in photoreceptor cells. Taken together, these results suggest that CLRN1 expression is potentially supported by a variety of retinal cells, and the right combination of AAV vector dose, promoter, and delivery method needs to be selected to develop safe therapies for USH3 disorder. Public Library of Science 2016-02-16 /pmc/articles/PMC4755610/ /pubmed/26881841 http://dx.doi.org/10.1371/journal.pone.0148874 Text en © 2016 Dinculescu et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Dinculescu, Astra
Stupay, Rachel M.
Deng, Wen-Tao
Dyka, Frank M.
Min, Seok-Hong
Boye, Sanford L.
Chiodo, Vince A.
Abrahan, Carolina E.
Zhu, Ping
Li, Qiuhong
Strettoi, Enrica
Novelli, Elena
Nagel-Wolfrum, Kerstin
Wolfrum, Uwe
Smith, W. Clay
Hauswirth, William W.
AAV-Mediated Clarin-1 Expression in the Mouse Retina: Implications for USH3A Gene Therapy
title AAV-Mediated Clarin-1 Expression in the Mouse Retina: Implications for USH3A Gene Therapy
title_full AAV-Mediated Clarin-1 Expression in the Mouse Retina: Implications for USH3A Gene Therapy
title_fullStr AAV-Mediated Clarin-1 Expression in the Mouse Retina: Implications for USH3A Gene Therapy
title_full_unstemmed AAV-Mediated Clarin-1 Expression in the Mouse Retina: Implications for USH3A Gene Therapy
title_short AAV-Mediated Clarin-1 Expression in the Mouse Retina: Implications for USH3A Gene Therapy
title_sort aav-mediated clarin-1 expression in the mouse retina: implications for ush3a gene therapy
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4755610/
https://www.ncbi.nlm.nih.gov/pubmed/26881841
http://dx.doi.org/10.1371/journal.pone.0148874
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