{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,11,11]],"date-time":"2026-11-11T21:43:52Z","timestamp":1794433432053,"version":"4.9.1"},"reference-count":52,"publisher":"MDPI AG","issue":"14","license":[{"start":{"date-parts":[[2023,7,12]],"date-time":"2023-07-12T00:00:00Z","timestamp":1689120000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000065","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["NS097726"],"award-info":[{"award-number":["NS097726"]}],"id":[{"id":"10.13039\/100000065","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000065","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["NS040337"],"award-info":[{"award-number":["NS040337"]}],"id":[{"id":"10.13039\/100000065","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Over a third of patients with temporal lobe epilepsy (TLE) are not effectively treated with current anti-seizure drugs, spurring the development of gene therapies. The injection of adeno-associated viral vectors (AAV) into the brain has been shown to be a safe and viable approach. However, to date, AAV expression of therapeutic genes has not been regulated. Moreover, a common property of antiepileptic drugs is a narrow therapeutic window between seizure control and side effects. Therefore, a long-term goal is to develop drug-inducible gene therapies that can be regulated by clinically relevant drugs. In this study, a first-generation doxycycline-regulated gene therapy that delivered an engineered version of the leak potassium channel Kcnk2 (TREK-M) was injected into the hippocampus of male rats. Rats were electrically stimulated until kindled. EEG was monitored 24\/7. Electrical kindling revealed an important side effect, as even low expression of TREK M in the absence of doxycycline was sufficient to cause rats to develop spontaneous recurring seizures. Treating the epileptic rats with doxycycline successfully reduced spontaneous seizures. Localization studies of infected neurons suggest seizures were caused by expression in GABAergic inhibitory neurons. In contrast, doxycycline increased the expression of TREK-M in excitatory neurons, thereby reducing seizures through net inhibition of firing. These studies demonstrate that drug-inducible gene therapies are effective in reducing spontaneous seizures and highlight the importance of testing for side effects with pro-epileptic stressors such as electrical kindling. These studies also show the importance of evaluating the location and spread of AAV-based gene therapies in preclinical studies.<\/jats:p>","DOI":"10.3390\/ijms241411347","type":"journal-article","created":{"date-parts":[[2023,7,13]],"date-time":"2023-07-13T01:43:22Z","timestamp":1689212602000},"page":"11347","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["Drug-Inducible Gene Therapy Effectively Reduces Spontaneous Seizures in Kindled Rats but Creates Off-Target Side Effects in Inhibitory Neurons"],"prefix":"10.3390","volume":"24","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-4621-5598","authenticated-orcid":false,"given":"Kyle A.","family":"Sullivan","sequence":"first","affiliation":[{"name":"Department of Pharmacology, University of Virginia, Charlottesville, VA 22980, USA"},{"name":"Computational and Predictive Biology, Oak Ridge National Laboratory, Oak Ridge, TN 37830, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Iuliia","family":"Vitko","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, University of Virginia, Charlottesville, VA 22980, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kathryn","family":"Blair","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, University of Virginia, Charlottesville, VA 22980, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ronald P.","family":"Gaykema","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, University of Virginia, Charlottesville, VA 22980, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6924-6610","authenticated-orcid":false,"given":"Madison J.","family":"Failor","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, University of Virginia, Charlottesville, VA 22980, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0002-4717-9450","authenticated-orcid":false,"given":"Jennifer M.","family":"San Pietro","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, University of Virginia, Charlottesville, VA 22980, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Deblina","family":"Dey","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, University of Virginia, Charlottesville, VA 22980, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"John M.","family":"Williamson","sequence":"additional","affiliation":[{"name":"Department of Neurology, University of Virginia, Charlottesville, VA 22980, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ruth L.","family":"Stornetta","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, University of Virginia, Charlottesville, VA 22980, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jaideep","family":"Kapur","sequence":"additional","affiliation":[{"name":"Department of Neurology, University of Virginia, Charlottesville, VA 22980, USA"},{"name":"UVA Brain Institute, University of Virginia, Charlottesville, VA 22980, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6237-3863","authenticated-orcid":false,"given":"Edward","family":"Perez-Reyes","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, University of Virginia, Charlottesville, VA 22980, USA"},{"name":"UVA Brain Institute, University of Virginia, Charlottesville, VA 22980, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,7,12]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"662","DOI":"10.1016\/j.wneu.2016.12.074","article-title":"Prevalence and Incidence of Drug-Resistant Mesial Temporal Lobe Epilepsy in the United States","volume":"99","author":"Stewart","year":"2017","journal-title":"World Neurosurg."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1069","DOI":"10.1111\/j.1528-1167.2009.02397.x","article-title":"Definition of drug resistant epilepsy: Consensus proposal by the ad hoc Task Force of the ILAE Commission on Therapeutic Strategies","volume":"51","author":"Kwan","year":"2010","journal-title":"Epilepsia"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"311","DOI":"10.1056\/NEJM200108023450501","article-title":"A randomized, controlled trial of surgery for temporal-lobe epilepsy","volume":"345","author":"Wiebe","year":"2001","journal-title":"N. 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