{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,23]],"date-time":"2026-08-23T18:07:54Z","timestamp":1787508474180,"version":"build-2736575974"},"reference-count":46,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2024,8,8]],"date-time":"2024-08-08T00:00:00Z","timestamp":1723075200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Cell. Neurosci."],"abstract":"<jats:sec>\n                    <jats:title>Introduction<\/jats:title>\n                    <jats:p>\n                      The\n                      <jats:italic>KCNQ2\/KCNQ3<\/jats:italic>\n                      genes encode the voltage-gated K channel underlying the neuronal M-current, regulating neuronal excitability. Loss-of-function (LoF) variants cause neonatal epilepsy, treatable with the M-current-opener retigabine, which is no longer marketed due to side effects. Gain-of-function (GoF) variants cause developmental encephalopathy and autism that could be amenable to M-current, but such therapies are not clinically available. In this translational project, we investigated whether donepezil, a cholinergic drug used in Alzheimer\u2019s, suppresses M currents\n                      <jats:italic>in vitro<\/jats:italic>\n                      and improves cognitive symptoms in patients with GoF variants.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      (1) The effect of 1\u2009\u03bcM donepezil on the amplitude of the M-current was measured in excitatory and inhibitory neurons of mouse primary cultured hippocampal cells. M-current was measured using the standard deactivation protocol (holding at 0\u2009mV and deactivation at \u221260\u2009mV) in the voltage-clamp configuration of the whole-cell patch clamp technique. The impact of donepezil was also examined on the spontaneous firing activity of hippocampal neurons in the current-clamp configuration. (2) Four children with autism, aged 2.5\u20138\u2009years, with the following GoF variants were enrolled:\n                      <jats:italic>KCNQ2 (p. Arg144Gln)<\/jats:italic>\n                      and\n                      <jats:italic>KCNQ 3 (p.Arg227Gln, p.Arg230Cys)<\/jats:italic>\n                      . Patients were treated off-label with donepezil 2.5\u20135\u2009mg\/d for 12\u2009months and assessed with: clinical Global Impression of Change (CGI-c), Childhood Autism Rating Scale 2 (CARS-2), Adaptive Behavior Assessment System-II (ABAS-II), and Child Development Inventory (CDI).\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      (1) Application of donepezil for at least 6\u2009min produced a significant inhibition of the M-current with an IC50 of 0.4\u2009\u03bcM. At 1\u2009\u03bcM, donepezil reduced by 67% the M-current density of excitatory neurons (2.4\u2009\u00b1\u20090.46 vs. 0.89\u2009\u00b1\u20090.15 pA\/pF,\n                      <jats:italic>p<\/jats:italic>\n                      \u2009&amp;lt;\u20090.05\n                      <jats:sup>*<\/jats:sup>\n                      ). In inhibitory neurons, application of 1\u2009\u03bcM donepezil produced a lesser inhibition of 59% of the M-current density (1.39\u2009\u00b1\u20090.43 vs. 0.57\u2009\u00b1\u20090.21,\n                      <jats:italic>p<\/jats:italic>\n                      \u2009&amp;gt;\u20090.05). Donepezil (1\u2009\u03bcM) potently increased by 2.6-fold the spontaneous firing frequency, which was prevented by the muscarinic receptor antagonist atropine (10\u2009\u03bcM). (2) The CARS-2 decreased by 3.8\u2009\u00b1\u20094.9 points (\n                      <jats:italic>p<\/jats:italic>\n                      \u2009&amp;gt;\u20090.05), but in two patients with\n                      <jats:italic>KCNQ3<\/jats:italic>\n                      variants, the improvement was over the 4.5 clinically relevant threshold. The global clinical change was also clinically significant in these patients (CGI-c\u2009=\u20091). The CDI increased by 65% (\n                      <jats:italic>p<\/jats:italic>\n                      \u2009&amp;lt;\u20090.05\n                      <jats:sup>*<\/jats:sup>\n                      ), while the ABAS-II remained unchanged.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Discussion<\/jats:title>\n                    <jats:p>\n                      Donepezil should be repurposed as a novel alternative treatment for GoF variants in\n                      <jats:italic>KCNQ2\/KCNQ3<\/jats:italic>\n                      encephalopathy.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.3389\/fncel.2024.1380442","type":"journal-article","created":{"date-parts":[[2024,8,8]],"date-time":"2024-08-08T00:27:54Z","timestamp":1723076874000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":10,"title":["Donepezil as a new therapeutic potential in KCNQ2- and KCNQ3-related autism"],"prefix":"10.3389","volume":"18","author":[{"given":"Andreea","family":"Nissenkorn","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lior","family":"Bar","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ariel","family":"Ben-Bassat","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lynn","family":"Rothstein","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hoda","family":"Abdelrahim","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Riki","family":"Sokol","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lidia V.","family":"Gabis","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bernard","family":"Attali","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2024,8,8]]},"reference":[{"key":"ref1","doi-asserted-by":"publisher","first-page":"972023","DOI":"10.3389\/fnmol.2022.972023","article-title":"Excitatory and inhibitory hippocampal neurons differ in their homeostatic adaptation to chronic M-channel modulation","volume":"15","author":"Bar","year":"2022","journal-title":"Front. 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