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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...

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Autores principales: 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
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society of Gene & Cell Therapy 2019
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.
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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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