{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,11]],"date-time":"2026-05-11T11:56:10Z","timestamp":1778500570996,"version":"3.51.4"},"reference-count":45,"publisher":"Public Library of Science (PLoS)","issue":"6","license":[{"start":{"date-parts":[[2022,6,16]],"date-time":"2022-06-16T00:00:00Z","timestamp":1655337600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Republic of Korea Ministry of SMEs and Startups","award":["TIPS (Korea Tech Incubator Program for Startup)"],"award-info":[{"award-number":["TIPS (Korea Tech Incubator Program for Startup)"]}]},{"DOI":"10.13039\/501100003725","name":"National Research Foundation of Korea","doi-asserted-by":"crossref","award":["NRF-2017R1A2B4012769"],"award-info":[{"award-number":["NRF-2017R1A2B4012769"]}],"id":[{"id":"10.13039\/501100003725","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/501100005006","name":"Asan Institute for Life Sciences, Asan Medical Center","doi-asserted-by":"publisher","award":["2019-287"],"award-info":[{"award-number":["2019-287"]}],"id":[{"id":"10.13039\/501100005006","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Republic of Korea Ministry of Health and Welfare","award":["HI15C2599"],"award-info":[{"award-number":["HI15C2599"]}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS ONE"],"abstract":"<jats:p>In addition to laser photocoagulation, therapeutic interventions for diabetic retinopathy (DR) have heretofore consisted of anti-VEGF drugs, which, besides drawbacks inherent to the treatments themselves, are limited in scope and may not fully address the condition\u2019s complex pathophysiology. This is because DR is a multifactorial condition, meaning a gene therapy focused on a target with broader effects, such as the mechanistic target of rapamycin (mTOR), may prove to be the solution in overcoming these concerns. Having previously demonstrated the potential of a mTOR-inhibiting shRNA packaged in a recombinant adeno-associated virus to address a variety of angiogenic retinal diseases, here we explore the effects of rAAV2-shmTOR-SD in a streptozotocin-induced diabetic mouse model. Delivered via intravitreal injection, the therapeutic efficacy of the virus vector upon early DR processes was examined. rAAV2-shmTOR-SD effectively transduced mouse retinas and therein downregulated mTOR expression, which was elevated in sham-treated and control shRNA-injected (rAAV2-shCon-SD) control groups. mTOR inhibition additionally led to marked reductions in pericyte loss, acellular capillary formation, vascular permeability, and retinal cell layer thinning, processes that contribute to DR progression. Immunohistochemistry showed that rAAV2-shmTOR-SD decreased ganglion cell loss and pathogenic M\u00fcller cell activation and proliferation, while also having anti-apoptotic activity, with these effects suggesting the therapeutic virus vector may be neuroprotective. Taken together, these results build upon our previous work to demonstrate the broad ability of rAAV2-shmTOR-SD to address aspects of DR pathophysiology further evidencing its potential as a human gene therapeutic strategy for DR.<\/jats:p>","DOI":"10.1371\/journal.pone.0269951","type":"journal-article","created":{"date-parts":[[2022,6,16]],"date-time":"2022-06-16T17:43:02Z","timestamp":1655401382000},"page":"e0269951","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":12,"title":["mTOR inhibition as a novel gene therapeutic strategy for diabetic retinopathy"],"prefix":"10.1371","volume":"17","author":[{"given":"Steven Hyun Seung","family":"Lee","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Joo Yong","family":"Lee","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jun-Sub","family":"Choi","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hee Jong","family":"Kim","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jin","family":"Kim","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Seho","family":"Cha","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kyoung Jin","family":"Lee","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ha-Na","family":"Woo","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Keerang","family":"Park","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4241-5664","authenticated-orcid":true,"given":"Heuiran","family":"Lee","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"340","published-online":{"date-parts":[[2022,6,16]]},"reference":[{"key":"pone.0269951.ref001","doi-asserted-by":"crossref","first-page":"164","DOI":"10.4103\/0974-9233.154389","article-title":"Novel pharmacotherapies in diabetic retinopathy","volume":"22","author":"VS Dedania","year":"2015","journal-title":"Middle East Afr J Ophthalmol"},{"key":"pone.0269951.ref002","doi-asserted-by":"crossref","first-page":"e93751","DOI":"10.1172\/jci.insight.93751","article-title":"Diabetic retinopathy: current understanding, mechanisms, and treatment strategies","volume":"2","author":"EJ Duh","year":"2017","journal-title":"JCI Insight"},{"key":"pone.0269951.ref003","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1042\/CS20120588","article-title":"Advances in our understanding of diabetic retinopathy","volume":"125","author":"AW Stitt","year":"2013","journal-title":"Clin Sci (Lond)"},{"key":"pone.0269951.ref004","doi-asserted-by":"crossref","first-page":"1816","DOI":"10.3390\/ijms19061816","article-title":"Diabetic retinopathy: pathophysiology and treatments","volume":"19","author":"W Wang","year":"2018","journal-title":"Int J Mol Sci"},{"key":"pone.0269951.ref005","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1007\/s00592-012-0449-3","article-title":"Pathophysiology and treatment of diabetic retinopathy","volume":"50","author":"F Bandello","year":"2013","journal-title":"Acta Diabetol"},{"key":"pone.0269951.ref006","doi-asserted-by":"crossref","first-page":"83","DOI":"10.1080\/08820538.2017.1353820","article-title":"Panretinal photocoagulation: a review of complications","volume":"33","author":"SV Reddy","year":"2018","journal-title":"Semin Ophthalmol"},{"key":"pone.0269951.ref007","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1016\/j.exer.2015.04.004","article-title":"Retinal fibrosis in diabetic retinopathy","volume":"142","author":"S Roy","year":"2016","journal-title":"Exp Eye Res"},{"key":"pone.0269951.ref008","doi-asserted-by":"crossref","first-page":"E21","DOI":"10.3390\/pharmaceutics10010021","article-title":"Extended duration vascular endothelial growth factor inhibition in the eye: failures, successes, and future possibilities","volume":"10","author":"MW Stewart","year":"2018","journal-title":"Pharmaceutics"},{"key":"pone.0269951.ref009","doi-asserted-by":"crossref","first-page":"329","DOI":"10.1016\/j.ajo.2011.05.040","article-title":"Emerging evidence concerning systemic safety of anti-VEGF agents\u2014should ophthalmologists be concerned?","volume":"152","author":"LS Lim","year":"2011","journal-title":"Am J Ophthalmol"},{"key":"pone.0269951.ref010","doi-asserted-by":"crossref","first-page":"2045","DOI":"10.1248\/bpb.b17-00475","article-title":"Mammalian target of rapamycin (mTOR) as a potential therapeutic target in pathological ocular angiogenesis","volume":"40","author":"T Nakahara","year":"2017","journal-title":"Biol Pharm Bull"},{"key":"pone.0269951.ref011","doi-asserted-by":"crossref","first-page":"2156273","DOI":"10.1155\/2016\/2156273","article-title":"Diabetic macular edema pathophysiology: vasogenic versus inflammatory","volume":"2016","author":"P Romero-Aroca","year":"2016","journal-title":"J Diabetes Res"},{"key":"pone.0269951.ref012","doi-asserted-by":"crossref","first-page":"7","DOI":"10.1016\/j.visres.2017.04.003","article-title":"The pathology associated with diabetic retinopathy","volume":"139","author":"J Lechner","year":"2017","journal-title":"Vision Res"},{"key":"pone.0269951.ref013","doi-asserted-by":"crossref","first-page":"960","DOI":"10.1016\/j.cell.2017.02.004","article-title":"mTOR signaling in growth, metabolism, and disease","volume":"168","author":"RA Saxton","year":"2017","journal-title":"Cell"},{"key":"pone.0269951.ref014","doi-asserted-by":"crossref","first-page":"e589813","DOI":"10.1155\/2011\/589813","article-title":"Potential therapeutic roles for inhibition of the PI3K\/Akt\/mTOR pathway in the pathophysiology of diabetic retinopathy","volume":"2011","author":"JL Jacot","year":"2011","journal-title":"J Ophthalmol"},{"key":"pone.0269951.ref015","doi-asserted-by":"crossref","first-page":"e88203","DOI":"10.1371\/journal.pone.0088203","article-title":"Temsirolimus inhibits proliferation and migration in retinal pigment epithelial and endothelial cells via mTOR inhibition and decreases VEGF and PDGF expression","volume":"9","author":"R Liegl","year":"2014","journal-title":"PLoS One"},{"key":"pone.0269951.ref016","doi-asserted-by":"crossref","first-page":"431","DOI":"10.1016\/j.bbrc.2009.04.143","article-title":"A novel program to design siRNAs simultaneously effective to highly variable virus genomes","volume":"384","author":"HS Lee","year":"2009","journal-title":"Biochem Biophys Res Commun"},{"key":"pone.0269951.ref017","doi-asserted-by":"crossref","first-page":"3147","DOI":"10.1007\/s00018-012-0998-1","article-title":"Multispecies-compatible antitumor effects of a cross-species small-interfering RNA against mammalian target of rapamycin","volume":"69","author":"J Ahn","year":"2012","journal-title":"Cell Mol Life Sci"},{"key":"pone.0269951.ref018","doi-asserted-by":"crossref","first-page":"2653","DOI":"10.2741\/2872","article-title":"Cancer gene therapy using adeno-associated virus vectors","volume":"13","author":"K Park","year":"2008","journal-title":"Front Biosci"},{"key":"pone.0269951.ref019","doi-asserted-by":"crossref","first-page":"45","DOI":"10.1167\/iovs.61.2.45","article-title":"Inhibition of mTOR via an AAV-delivered shRNA tested in a rat OIR model as a potential antiangiogenic gene therapy","volume":"61","author":"SHS Lee","year":"2020","journal-title":"Invest Ophthalmol Vis Sci"},{"key":"pone.0269951.ref020","doi-asserted-by":"crossref","first-page":"553","DOI":"10.1007\/s12015-016-9670-8","article-title":"Cellular GFP toxicity and immunogenicity: potential confounders in in vivo cell tracking experiments","volume":"12","author":"AM Ansari","year":"2016","journal-title":"Stem Cell Rev"},{"key":"pone.0269951.ref021","doi-asserted-by":"crossref","first-page":"291","DOI":"10.1165\/rcmb.2014-0338MA","article-title":"Modeling pulmonary disease pathways using recombinant adeno-associated virus 6.2","volume":"53","author":"B Strobel","year":"2015","journal-title":"Am J Respir Cell Mol Biol"},{"key":"pone.0269951.ref022","doi-asserted-by":"crossref","first-page":"171","DOI":"10.1016\/j.omtm.2019.06.004","article-title":"Effects of stuffer DNA on the suppression of choroidal neovascularization by a rAAV expressing a mTOR-inhibiting shRNA","volume":"14","author":"SHS Lee","year":"2019","journal-title":"Mol Ther Methods Clin Dev"},{"key":"pone.0269951.ref023","doi-asserted-by":"crossref","first-page":"444","DOI":"10.1242\/dmm.009597","article-title":"Update on animal models of diabetic retinopathy: from molecular approaches to mice and higher mammals","volume":"5","author":"R Robinson","year":"2012","journal-title":"Dis Model Mech"},{"key":"pone.0269951.ref024","doi-asserted-by":"crossref","first-page":"26","DOI":"10.1016\/j.omtn.2017.05.012","article-title":"Adeno-associated viral vector-mediated mTOR inhibition by short hairpin RNA suppresses laser-induced choroidal neovascularization","volume":"8","author":"TK Park","year":"2017","journal-title":"Mol Ther Nucleic Acids"},{"key":"pone.0269951.ref025","doi-asserted-by":"crossref","first-page":"5.47.1","DOI":"10.1002\/0471141755.ph0547s70","article-title":"Streptozotocin-induced diabetic models in mice and rats","volume":"70","author":"BL Furman","year":"2015","journal-title":"Curr Protoc Pharmacol"},{"key":"pone.0269951.ref026","first-page":"e50489","article-title":"Trypsin digest protocol to analyze the retinal vasculature of a mouse model","volume":"76","author":"JC Chou","year":"2013","journal-title":"J Vis Exp"},{"key":"pone.0269951.ref027","doi-asserted-by":"crossref","first-page":"1257","DOI":"10.1080\/14712598.2018.1545836","article-title":"Diabetic retinopathy: a complex pathophysiology requiring novel therapeutic strategies","volume":"18","author":"M Whitehead","year":"2018","journal-title":"Expert Opin Biol Ther"},{"key":"pone.0269951.ref028","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1080\/10739680601072123","article-title":"Arteriolar involvement in the microvascular lesions of diabetic retinopathy: implications for pathogenesis","volume":"14","author":"TA Gardiner","year":"2007","journal-title":"Microcirculation"},{"key":"pone.0269951.ref029","first-page":"E2655","article-title":"Retinal neurodegeneration may precede microvascular changes characteristic of diabetic retinopathy in diabetes mellitus","volume":"113","author":"EH Sohn","year":"2016","journal-title":"Proc Natl Acad Sci U S A"},{"key":"pone.0269951.ref030","doi-asserted-by":"crossref","first-page":"942","DOI":"10.3390\/ijms19040942","article-title":"Role of inflammation in diabetic retinopathy","volume":"19","author":"A R\u00fcbsam","year":"2018","journal-title":"Int J Mol Sci"},{"key":"pone.0269951.ref031","doi-asserted-by":"crossref","first-page":"4356","DOI":"10.1167\/iovs.16-19197","article-title":"Defective autophagy in diabetic retinopathy","volume":"57","author":"JM Lopes de Faria","year":"2016","journal-title":"Invest Ophthalmol Vis Sci"},{"key":"pone.0269951.ref032","doi-asserted-by":"crossref","first-page":"2297","DOI":"10.2337\/db09-1420","article-title":"M\u00fcller cell-derived VEGF is essential for diabetes-induced retinal inflammation and vascular leakage","volume":"59","author":"J Wang","year":"2010","journal-title":"Diabetes"},{"key":"pone.0269951.ref033","first-page":"1971","article-title":"Glial reactivity, an early feature of diabetic retinopathy","volume":"41","author":"E Rungger-Br\u00e4ndle","year":"2000","journal-title":"Invest Ophthalmol Vis Sci"},{"key":"pone.0269951.ref034","doi-asserted-by":"crossref","first-page":"783","DOI":"10.1172\/JCI2425","article-title":"Neural apoptosis in the retina during experimental and human diabetes: early onset and effect of insulin","volume":"102","author":"AJ Barber","year":"1998","journal-title":"J Clin Invest"},{"key":"pone.0269951.ref035","doi-asserted-by":"crossref","first-page":"82","DOI":"10.1016\/j.visres.2017.06.014","article-title":"Neurodegeneration in diabetic retinopathy: potential for novel therapies","volume":"139","author":"AJ Barber","year":"2017","journal-title":"Vision Res"},{"key":"pone.0269951.ref036","doi-asserted-by":"crossref","first-page":"463","DOI":"10.1017\/S0952523800173122","article-title":"Neuronal and microglial response in the retina of streptozotocin-induced diabetic rats","volume":"17","author":"XX Zeng","year":"2000","journal-title":"Vis Neurosci"},{"key":"pone.0269951.ref037","doi-asserted-by":"crossref","first-page":"eaar3938","DOI":"10.1126\/sciadv.aar3938","article-title":"ETV7 is an essential component of rapamycin-insensitive mTOR complex in cancer","volume":"4","author":"FC Harwood","year":"2018","journal-title":"Sci Adv"},{"key":"pone.0269951.ref038","doi-asserted-by":"crossref","first-page":"1570","DOI":"10.1159\/000453207","article-title":"Blocking mammalian target of rapamycin (mTOR) attenuates HIF-1\u03b1 pathways engaged-vascular endothelial growth factor (VEGF) in diabetic retinopathy","volume":"40","author":"J Wei","year":"2016","journal-title":"Cell Physiol Biochem"},{"key":"pone.0269951.ref039","doi-asserted-by":"crossref","first-page":"28","DOI":"10.1159\/000431129","article-title":"Ocular angiogenesis: vascular endothelial growth factor and other factors","volume":"55","author":"RG Rubio","year":"2016","journal-title":"Dev Ophthalmol"},{"key":"pone.0269951.ref040","doi-asserted-by":"crossref","first-page":"113","DOI":"10.1007\/s11010-013-1760-1","article-title":"Concerted inhibition of HIF-1\u03b1 and -2\u03b1 expression markedly suppresses angiogenesis in cultured RPE cells","volume":"383","author":"T Nakajima","year":"2013","journal-title":"Mol Cell Biochem"},{"key":"pone.0269951.ref041","doi-asserted-by":"crossref","first-page":"681","DOI":"10.1016\/j.molcel.2012.09.017","article-title":"HIF2\u03b1 acts as an mTORC1 activator through the amino acid carrier SLC7A5","volume":"48","author":"A Elorza","year":"2012","journal-title":"Mol Cell"},{"key":"pone.0269951.ref042","doi-asserted-by":"crossref","first-page":"4665","DOI":"10.1096\/fj.201700336R","article-title":"Animal models of ocular angiogenesis: from development to pathologies","volume":"31","author":"CH Liu","year":"2017","journal-title":"FASEB J"},{"key":"pone.0269951.ref043","doi-asserted-by":"crossref","first-page":"500","DOI":"10.1016\/j.preteyeres.2010.05.003","article-title":"Animal models of choroidal and retinal neovascularization","volume":"29","author":"HE Grossniklaus","year":"2010","journal-title":"Prog Retin Eye Res"},{"key":"pone.0269951.ref044","doi-asserted-by":"crossref","first-page":"93","DOI":"10.1007\/s11892-017-0913-0","article-title":"Animal models of diabetic retinopathy","volume":"17","author":"AM Olivares","year":"2017","journal-title":"Curr Diab Rep"},{"key":"pone.0269951.ref045","doi-asserted-by":"crossref","first-page":"1450","DOI":"10.1096\/fj.03-1476fje","article-title":"A central role for inflammation in the pathogenesis of diabetic retinopathy","volume":"18","author":"AM Joussen","year":"2004","journal-title":"FASEB J"}],"container-title":["PLOS ONE"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0269951","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,2,8]],"date-time":"2023-02-08T16:39:21Z","timestamp":1675874361000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0269951"}},"subtitle":[],"editor":[{"given":"Zhijian","family":"Wu","sequence":"first","affiliation":[],"role":[{"role":"editor","vocabulary":"crossref"}]}],"short-title":[],"issued":{"date-parts":[[2022,6,16]]},"references-count":45,"journal-issue":{"issue":"6","published-online":{"date-parts":[[2022,6,16]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.pone.0269951","relation":{},"ISSN":["1932-6203"],"issn-type":[{"value":"1932-6203","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,6,16]]}}}