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Effects of Stuffer DNA on the Suppression of Choroidal Neovascularization by a rAAV Expressing a mTOR-Inhibiting shRNA
Choroidal neovascularization (CNV) is the defining characteristic of the wet subtype of age-related macular degeneration (AMD), which is a rapidly growing global health problem. Previously, we had demonstrated the therapeutic potential of gene therapy against CNV using short hairpin RNA (shRNA) deli...
Autores principales: | , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Society of Gene & Cell Therapy
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6661460/ https://www.ncbi.nlm.nih.gov/pubmed/31380463 http://dx.doi.org/10.1016/j.omtm.2019.06.004 |
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author | Lee, Steven Hyun Seung Chang, HeeSoon Kim, Hee Jong Choi, Jun-Sub Kim, Jin Kim, Ji Hyun Woo, Ha-Na Nah, Seung Kwan Jung, Sang Joon Lee, Joo Yong Park, Keerang Park, Tae Kwann Lee, Heuiran |
author_facet | Lee, Steven Hyun Seung Chang, HeeSoon Kim, Hee Jong Choi, Jun-Sub Kim, Jin Kim, Ji Hyun Woo, Ha-Na Nah, Seung Kwan Jung, Sang Joon Lee, Joo Yong Park, Keerang Park, Tae Kwann Lee, Heuiran |
author_sort | Lee, Steven Hyun Seung |
collection | PubMed |
description | Choroidal neovascularization (CNV) is the defining characteristic of the wet subtype of age-related macular degeneration (AMD), which is a rapidly growing global health problem. Previously, we had demonstrated the therapeutic potential of gene therapy against CNV using short hairpin RNA (shRNA) delivered via recombinant adeno-associated virus (rAAV), which abrogates mammalian-to-mechanistic (mTOR) activity in a novel manner by simultaneously inhibiting both mTOR complexes. Both the target and use of gene therapy represent a novel treatment modality against AMD. Here, the xenogeneic GFP gene used as a reporter in previous studies was removed from the virus vector to further develop the therapeutic for clinical trials. Instead, a stuffer DNA derived from the 3′ UTR of the human UBE3A gene was used to ensure optimal viral genome size for efficient rAAV assembly. The virus vector containing the stuffer DNA, rAAV2-shmTOR-SD, positively compares to one encoding the shRNA and a GFP expression cassette in terms of reducing CNV in a laser-induced mouse model, as determined by fundus fluorescein angiography. These results were confirmed via immunohistochemistry using anti-CD31, while a TUNEL assay showed that rAAV2-shmTOR-SD possesses anti-apoptotic properties as well. The qualities exhibited by rAAV2-shmTOR-SD demonstrate its potential as a human gene therapeutic for the treatment of wet AMD. |
format | Online Article Text |
id | pubmed-6661460 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | American Society of Gene & Cell Therapy |
record_format | MEDLINE/PubMed |
spelling | pubmed-66614602019-08-02 Effects of Stuffer DNA on the Suppression of Choroidal Neovascularization by a rAAV Expressing a mTOR-Inhibiting shRNA Lee, Steven Hyun Seung Chang, HeeSoon Kim, Hee Jong Choi, Jun-Sub Kim, Jin Kim, Ji Hyun Woo, Ha-Na Nah, Seung Kwan Jung, Sang Joon Lee, Joo Yong Park, Keerang Park, Tae Kwann Lee, Heuiran Mol Ther Methods Clin Dev Article Choroidal neovascularization (CNV) is the defining characteristic of the wet subtype of age-related macular degeneration (AMD), which is a rapidly growing global health problem. Previously, we had demonstrated the therapeutic potential of gene therapy against CNV using short hairpin RNA (shRNA) delivered via recombinant adeno-associated virus (rAAV), which abrogates mammalian-to-mechanistic (mTOR) activity in a novel manner by simultaneously inhibiting both mTOR complexes. Both the target and use of gene therapy represent a novel treatment modality against AMD. Here, the xenogeneic GFP gene used as a reporter in previous studies was removed from the virus vector to further develop the therapeutic for clinical trials. Instead, a stuffer DNA derived from the 3′ UTR of the human UBE3A gene was used to ensure optimal viral genome size for efficient rAAV assembly. The virus vector containing the stuffer DNA, rAAV2-shmTOR-SD, positively compares to one encoding the shRNA and a GFP expression cassette in terms of reducing CNV in a laser-induced mouse model, as determined by fundus fluorescein angiography. These results were confirmed via immunohistochemistry using anti-CD31, while a TUNEL assay showed that rAAV2-shmTOR-SD possesses anti-apoptotic properties as well. The qualities exhibited by rAAV2-shmTOR-SD demonstrate its potential as a human gene therapeutic for the treatment of wet AMD. American Society of Gene & Cell Therapy 2019-07-03 /pmc/articles/PMC6661460/ /pubmed/31380463 http://dx.doi.org/10.1016/j.omtm.2019.06.004 Text en © 2019 The Authors 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 Lee, Steven Hyun Seung Chang, HeeSoon Kim, Hee Jong Choi, Jun-Sub Kim, Jin Kim, Ji Hyun Woo, Ha-Na Nah, Seung Kwan Jung, Sang Joon Lee, Joo Yong Park, Keerang Park, Tae Kwann Lee, Heuiran Effects of Stuffer DNA on the Suppression of Choroidal Neovascularization by a rAAV Expressing a mTOR-Inhibiting shRNA |
title | Effects of Stuffer DNA on the Suppression of Choroidal Neovascularization by a rAAV Expressing a mTOR-Inhibiting shRNA |
title_full | Effects of Stuffer DNA on the Suppression of Choroidal Neovascularization by a rAAV Expressing a mTOR-Inhibiting shRNA |
title_fullStr | Effects of Stuffer DNA on the Suppression of Choroidal Neovascularization by a rAAV Expressing a mTOR-Inhibiting shRNA |
title_full_unstemmed | Effects of Stuffer DNA on the Suppression of Choroidal Neovascularization by a rAAV Expressing a mTOR-Inhibiting shRNA |
title_short | Effects of Stuffer DNA on the Suppression of Choroidal Neovascularization by a rAAV Expressing a mTOR-Inhibiting shRNA |
title_sort | effects of stuffer dna on the suppression of choroidal neovascularization by a raav expressing a mtor-inhibiting shrna |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6661460/ https://www.ncbi.nlm.nih.gov/pubmed/31380463 http://dx.doi.org/10.1016/j.omtm.2019.06.004 |
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