TREK channel activation suppresses migraine pain phenotype

•Trek1 and Trek2 invalidation increases TG excitability generates migraine•TREK1 TREK2 activation prevents migraine in rodents•TREK1/TREK2 agonists are as good the most efficient current cares for migraine•Targeting TREK channels needs to be considered clinical treatment on Activation sensitization of trigeminal ganglia (TG) sensory neurons, leading release pro-inflammatory peptides such calcitonin gene-related peptide (CGRP), likely a key component migraine-related headache induction. Reducing neuron represents therefore an attractive alternative strategy relieve pain. Here by using pharmacology genetic ex vivo vivo, we demonstrate that activating TREK1 two-pore-domain potassium (K2P) inhibits neuronal firing sufficiently fully reverse migraine-like phenotype induced NO-donors rodents. Finally, targeting is with CGRP antagonists, which one effective therapies. Altogether, our results inhibiting pharmacological should treatment. Migraine disabling conditions worldwide, affecting around 15% global population (Burch et al., 2015Burch R.C. Loder S. E. Smitherman T.A. The prevalence burden severe United States: updated statistics from government health surveillance studies.Headache. 2015; 55: 21-34Crossref PubMed Scopus (306) Google Scholar). It manifests unilateral throbbing accompanied multiple symptoms nausea, vomiting, photophobia, phonophobia, or cutaneous allodynia. generally assumed attacks start neurons (Strassman 1996Strassman A.M. Raymond S.A. Burstein R. Sensitization meningeal origin headaches.Nature. 1996; 384: 560-564Crossref (514) Hyperexcitability fibers innervating meninges induces neuropeptides (e.g. substance P [CGRP]) triggers vasodilatation surrounding blood vessels local neurogenic inflammation, (Frederiksen 2019Frederiksen S.D. Haanes K.A. Warfvinge K. Edvinsson L. Perivascular neurotransmitters: regulation cerebral flow role primary headaches.J. Cereb. Blood Flow Metab. 2019; 39: 610-632Crossref (20) Scholar; Goadsby 2017Goadsby P.J. Holland P.R. Martins-Oliveira M. Hoffmann J. Schankin C. Akerman Pathophysiology migraine: Disorder Processing.Physiol. Rev. 2017; 97: 553-622Crossref (621) Khan 2019Khan Amin F.M. Christensen C.E. Ghanizada H. Younis Olinger A.C.R. de Koning P.J.H. Larsson H.B.W. Ashina Meningeal contribution pain: magnetic resonance angiography study.Brain. 142: 93-102Crossref (38) Targeting secreted constitutes treat (Edvinsson 2018Edvinsson Krause D.N. target new therapies - successful translation bench clinic.Nat. Neurol. 2018; 14: 338-350Crossref (329) However, about 50% patients do not respond this suffer secondary effects (joint pain, dizziness, constipation, flu-like symptoms) patient care developed. Expression several K2P channel subunits has been detected nociceptive dorsal root ganglions (Alloui 2006Alloui A. Zimmermann Mamet Duprat F. Noël Chemin Guy N. Blondeau Voilley Rubat-Coudert al.TREK-1, K+ involved polymodal pain perception.EMBO 2006; 25: 2368-2376Crossref (301) Yamamoto 2009Yamamoto Y. Hatakeyama T. Taniguchi Immunohistochemical colocalization TREK-1, TREK-2 TRAAK TRP ganglion cells.Neurosci. Lett. 2009; 454: 129-133Crossref (52) We recently found human mutation K2P-TRESK channel, “TRESK-MT”, related migraine, K2P-TREK1 generate (Royal 2019Royal P. Andres-Bilbe Ávalos Prado Verkest Wdziekonski B. Schaub Baron Lesage Gasull X. Levitz Sandoz G. Migraine-associated TRESK mutations increase through Initiation inhibition TREK.Neuron. 101: 232-245.e6Abstract Full Text PDF (59) These importance excitability. As drugs against neuropeptide released its receptor Scholar), it act prevent and, consequently, migraine. made hypothesis enhancing activity would reduce excitability, peptides, suppress show that, at cellular level, ML67-33, TREK1/2 agonist (Bagriantsev 2013Bagriantsev S.N. Ang K.H. Gallardo-Godoy Clark Arkin M.R. Renslo A.R. Minor D.L. A high-throughput functional screen identifies small molecule regulators temperature- mechano-sensitive channels.ACS Chem. Biol. 2013; 8: 1841-1851Crossref (68) Scholar) reduces explaining “anti-migraine” properties. Using behavioral tests wild-type double knock-out mice Trek1/Trek2 (Trek1−/−-Trek2−/− mice), ML67-33 reverses NO donor-induced phenotype. This reversion exhibits similar potency BIBN4096, selective antagonist used (Olesen 2004Olesen Diener H.C. Husstedt I.W. Hall D. Meier U. Pollentier Lesko L.M. BIBN 4096 BS Clinical Proof Concept Study Group Calcitonin acute migraine.N. Engl. Med. 2004; 350: 1104-1110Crossref (987) completely reversed facial allodynia rats, direct index Therefore, inhibit underlying when developing molecules To investigate whether first verified specifically activated channels. validated selectively both homodimer, well TRESK-TREK1 heterodimer currents, whereas alone was (Figure S1). shows binding only moiety, can agonize channels, making choice studying homo- function. Once validated, next monitored well-described model chronic nitric oxide (NO) donors (Bates 2010Bates E.A. Nikai Brennan K.C. Fu Y.H. Charles A.C. Basbaum A.I. Ptácek L.J. Ahn A.H. Sumatriptan alleviates nitroglycerin-induced mechanical thermal mice.Cephalalgia. 2010; 30: 170-178Crossref (126) isosorbide dinitrate (ISDN) (Dallel 2018Dallel Descheemaeker Luccarini Recurrent administration donor, dinitrate, persistent cephalic hypersensitivity: progression.Cephalalgia. 38: 776-785Crossref (16) Royal 2018Verkest Piquet Diochot Dauvois Lanteri-Minet Lingueglia Effects systemic inhibitors acid-sensing ion 1 (ASIC1) rodent migraine.Br. Pharmacol. 175: 4154-4166Crossref (22) evaluated allodynia, quantifiable marker after four consecutive days ISDN injections Trek1−/−-Trek2−/− measuring paw withdrawal threshold dynamic von Frey esthesiometer before each injection 1A). measured consequences threshold. In animals, observed significant decrease compared last measurements (4.04 ± 0.06 g vs 2.87 0.04 g, p < 0.001). already exhibited low threshold, previously reported 3.11 0.08 i.p., respectively, > 0.001) 1B). then effect challenged capacity lowered injections. For purpose, were classified into three groups (for each), following series two different spaced hours time (Figures 1D 1F). group injected vehicle solution (saline + DMSO 0.1%) BIBN4096. second BIBN4096 solution. third efficacy confirmed group, since strongly increased their (2.92 0.10 3.47 0.11 2.79 0.13 3.28 0.26 mice, 0.2 conditions) 1F, black graphs). rapidly animals (2.89 0.07 3.61 0.01; 0.18 3.87 0.17 0.001, respectively) 1C 1E, blue traces), no additive (3.61 3.85 3.82 Nevertheless, responded differently, control (2.78 3.92 0.18, 1C, red trace), had impact (2.69 3.00 0.16, 0.1) trace). Interestingly, BIBN67-33 (3.92 3.65 0.19, 1D, graph), while (3.00 0.16 0.05) graph). Together, these indicate either antagonizing receptors BIBN4098 similarly NO-donors-induced mice. regulating currents studied medium-sized KO animals. classically divided within populations, according ability bind plant lectin isolectin B4 (IB4) Griffonia simplicifolia (Stucky Lewin, 1999Stucky C.L. Lewin G.R. Isolectin B(4)-positive -negative nociceptors functionally distinct.J. Neurosci. 1999; 19: 6497-6505Crossref Both IB4+ IB4- nociception but enriched neuropeptides, notably CGRP, known inducing inflammation triggering On cultured perfusion amplitude sustained (3.18 0.46 pA/pF 7.66 1.80 exposure respectively 0.05; 2.19 0.30 8.12 1.62 0.01, −25 mV) 2A 2B ). difference obtained (4.71 1.75 5.05 0.76 IB4+, 0.3; 2.77 0.31 3.60 0.29 IB4-, 0.05, 2C 2D). activates and/or neurons. determined TREK1-TREK2 First, showed Trek1 deletion 3 A-D, white points gray traces; 1.70 0.33 Hz 5.12 1.52 Trek1−/−-Trek2−/−, 10.89 1.57 150 pA stimulus), indicating required Second, led non-significant spike frequency (1.70 1.20 0.36 Hz, 0.2) 3A) important reduction (4.80 0.90 1.00 3B). ML67-33-induced might because rest rate (Guo Cao, 2014Guo Z. Cao Y.Q. Over-expression K(+) nociceptors.PLoS One. 2014; 9: e87029Crossref (39) Guo Liu Ren Nonmigraine-associated variant C110R does neurons.J. Neurophysiol. 112: 568-579Crossref specific (IB4+: 1.51 4.50 1.19 perfusion, IB4-: 9.67 3.03 respectively; 0.6) 3C 3D). Furthermore, rheobase (i.e. lowest intensity necessary action potentials (APs) significantly enhanced S2). indicates Excitability closely linked resting membrane potential (RMP). Because serve hub RMP maintenance negative values (Enyedi Czirják, 2010Enyedi Czirják Molecular background leak currents: two-pore domain channels.Physiol. 90: 559-605Crossref (558) parameter. hyperpolarization 3E S3, −51.32 2.40 mV −62.13 2.84 mV, 0.001; −53.96 0.91 −61.46 2.21 0.01) mainly slight statistically (−49.94 1.42 −53.94 2.28 0.2; −45.70 1.12 −50.70 1.81 0.06). addition, noticed basal value peptidergic higher than those (−45.70 3G). confirm rest, value. data play crucial particularly determining generation APs setting up close equilibrium potential. Thus, treated NO-donors. validate decreases suppresses NO-migraine conducted experiments rats. allows measure face reliable readout (Harris 2017Harris H.M. Carpenter J.M. Black J.R. Sufka K.J. repeated nitroglycerin administrations rats; modeling endpoints chronification.J. Methods. 284: 63-70Crossref (29) Pradhan 2014Pradhan A.A. Smith M.L. McGuire Tarash I. Evans C.J. Characterization novel migraine.Pain. 155: 269-274Abstract (125) protocol induce despite use sensitivity instead readout. After week habituation, tested filaments. Migraine-like daily intraperitoneally (i.p.) (10 mg/kg) over days. At day 5, administrated followed hours, every 30 min 4A). described (Verkest strong normal (p 0.001 groups) 4B). associated development reflects induction hyperexcitability shown Figure 4C, treatments manner (5.7 0.77 6.7 0.39 rats 4C 4D). models efficiently neurological disease caused combination environmental, hormonal, components. Genome-wide association studies have revealed synaptic transmission predisposition (Gormley 2016Gormley Anttila V. Winsvold B.S. Palta Esko Pers T.H. Farh Cuenca-Leon Muona Furlotte N.A. al.Meta-analysis 375,000 individuals 38 susceptibility loci migraine.Nat. Genet. 2016; 48: 856-866Crossref (286) Nyholt 2008Nyholt D.R. LaForge K.S. Kallela Alakurtti Färkkilä Hämaläinen Kaprio Kaunisto M.A. Heath al.A high-density 155 transport genes involvement common migraine.Hum. Mol. 2008; 17: 3318-3331Crossref (76) Particularly, fundamental disease. frameshift TRESK-MT perfectly segregates large pedigree (Andres-Enguix 2012Andres-Enguix Shang Stansfeld Morahan Sansom M.S. Lafrenière R.G. Roy Griffiths L.R. Rouleau G.A. Ebers G.C. al.Functional analysis missense variants (KCNK18) K channel.Sci. Rep. 2012; 2: 237Crossref (65) 2010Lafrenière Cader M.Z. Poulin J.F. Andres-Enguix Simoneau Gupta Boisvert McLaughlan Dubé M.P. dominant-negative familial aura.Nat. 16: 1157-1160Crossref (247) causality linking demonstrated showing CRISPR-Cas9 fix MT restores nociceptor (Pettingill 2019Pettingill Weir Wei Wu Flower Lalic Handel Duggal Chintawar Cheung causal loss function mechanisms.Brain. 3852-3867Crossref (23) generated altered proteins affect fragment binds makes candidates subsequently analyze modulating disease, combined approach deficient Trek2. (i) (ii) effect. fact TRAAK, also modulated did restore ISDN-induce ruling out participation due activation. trigger Trek1/2 deletion. observations suggest deleting promote phenotype, further rat face, directly state Googl