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Development and biological characterization of a clinical gene transfer vector for the treatment of MAK-associated retinitis pigmentosa
By combining next generation whole exome sequencing and induced pluripotent stem cell (iPSC) technology we found that an Alu repeat inserted in exon 9 of the MAK gene results in a loss of normal MAK transcript and development of human autosomal recessive retinitis pigmentosa (RP). Although a relativ...
Autores principales: | , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9159943/ https://www.ncbi.nlm.nih.gov/pubmed/34518651 http://dx.doi.org/10.1038/s41434-021-00291-5 |
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author | Tucker, Budd A. Burnight, Erin R. Cranston, Cathryn M. Ulferts, Mallory J. Luse, Meagan A. Westfall, Trudi Scott, C. Anthony Marsden, Autumn Gibson-Corley, Katherine Wiley, Luke A. Han, Ian C. Slusarski, Diane C. Mullins, Robert F. Stone, Edwin M. |
author_facet | Tucker, Budd A. Burnight, Erin R. Cranston, Cathryn M. Ulferts, Mallory J. Luse, Meagan A. Westfall, Trudi Scott, C. Anthony Marsden, Autumn Gibson-Corley, Katherine Wiley, Luke A. Han, Ian C. Slusarski, Diane C. Mullins, Robert F. Stone, Edwin M. |
author_sort | Tucker, Budd A. |
collection | PubMed |
description | By combining next generation whole exome sequencing and induced pluripotent stem cell (iPSC) technology we found that an Alu repeat inserted in exon 9 of the MAK gene results in a loss of normal MAK transcript and development of human autosomal recessive retinitis pigmentosa (RP). Although a relatively rare cause of disease in the general population, the MAK variant is enriched in individuals of Jewish ancestry. In this population, 1 in 55 individuals are carriers and one third of all cases of recessive RP is caused by this gene. The purpose of this study was to determine if a viral gene augmentation strategy could be used to safely restore functional MAK protein as a step toward a treatment for early stage MAK-associated RP. Patient iPSC-derived photoreceptor precursor cells were generated and transduced with viral vectors containing the MAK transcript. One week after transduction, transcript and protein could be detected via rt-PCR and western blotting respectively. Using patient-derived fibroblast cells and mak knockdown zebra fish we demonstrate that over-expression of the retinal MAK transgene restored the cells ability to regulate primary cilia length. In addition, the visual defect in mak knockdown zebrafish was mitigated via treatment with the retinal MAK transgene. There was no evidence of local or systemic toxicity at 1-month or 3-months following subretinal delivery of clinical grade vector into wild type rats. The findings reported here will help pave the way for initiation of a phase 1 clinical trial for the treatment of patients with MAK-associated RP. |
format | Online Article Text |
id | pubmed-9159943 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-91599432022-06-03 Development and biological characterization of a clinical gene transfer vector for the treatment of MAK-associated retinitis pigmentosa Tucker, Budd A. Burnight, Erin R. Cranston, Cathryn M. Ulferts, Mallory J. Luse, Meagan A. Westfall, Trudi Scott, C. Anthony Marsden, Autumn Gibson-Corley, Katherine Wiley, Luke A. Han, Ian C. Slusarski, Diane C. Mullins, Robert F. Stone, Edwin M. Gene Ther Article By combining next generation whole exome sequencing and induced pluripotent stem cell (iPSC) technology we found that an Alu repeat inserted in exon 9 of the MAK gene results in a loss of normal MAK transcript and development of human autosomal recessive retinitis pigmentosa (RP). Although a relatively rare cause of disease in the general population, the MAK variant is enriched in individuals of Jewish ancestry. In this population, 1 in 55 individuals are carriers and one third of all cases of recessive RP is caused by this gene. The purpose of this study was to determine if a viral gene augmentation strategy could be used to safely restore functional MAK protein as a step toward a treatment for early stage MAK-associated RP. Patient iPSC-derived photoreceptor precursor cells were generated and transduced with viral vectors containing the MAK transcript. One week after transduction, transcript and protein could be detected via rt-PCR and western blotting respectively. Using patient-derived fibroblast cells and mak knockdown zebra fish we demonstrate that over-expression of the retinal MAK transgene restored the cells ability to regulate primary cilia length. In addition, the visual defect in mak knockdown zebrafish was mitigated via treatment with the retinal MAK transgene. There was no evidence of local or systemic toxicity at 1-month or 3-months following subretinal delivery of clinical grade vector into wild type rats. The findings reported here will help pave the way for initiation of a phase 1 clinical trial for the treatment of patients with MAK-associated RP. Nature Publishing Group UK 2021-09-14 2022 /pmc/articles/PMC9159943/ /pubmed/34518651 http://dx.doi.org/10.1038/s41434-021-00291-5 Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Article Tucker, Budd A. Burnight, Erin R. Cranston, Cathryn M. Ulferts, Mallory J. Luse, Meagan A. Westfall, Trudi Scott, C. Anthony Marsden, Autumn Gibson-Corley, Katherine Wiley, Luke A. Han, Ian C. Slusarski, Diane C. Mullins, Robert F. Stone, Edwin M. Development and biological characterization of a clinical gene transfer vector for the treatment of MAK-associated retinitis pigmentosa |
title | Development and biological characterization of a clinical gene transfer vector for the treatment of MAK-associated retinitis pigmentosa |
title_full | Development and biological characterization of a clinical gene transfer vector for the treatment of MAK-associated retinitis pigmentosa |
title_fullStr | Development and biological characterization of a clinical gene transfer vector for the treatment of MAK-associated retinitis pigmentosa |
title_full_unstemmed | Development and biological characterization of a clinical gene transfer vector for the treatment of MAK-associated retinitis pigmentosa |
title_short | Development and biological characterization of a clinical gene transfer vector for the treatment of MAK-associated retinitis pigmentosa |
title_sort | development and biological characterization of a clinical gene transfer vector for the treatment of mak-associated retinitis pigmentosa |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9159943/ https://www.ncbi.nlm.nih.gov/pubmed/34518651 http://dx.doi.org/10.1038/s41434-021-00291-5 |
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