title: Inhibition of the kinase WNK1/HSN2 ameliorates neuropathic pain by restoring GABA inhibition.
abstract: HSN2is a nervous system predominant exon of the gene encoding the kinase WNK1 and is mutated in an autosomal recessive, inherited form of congenital pain insensitivity. The HSN2-containing splice variant is referred to as WNK1/HSN2. We created a knockout mouse specifically lacking theHsn2exon ofWnk1 Although these mice had normal spinal neuron and peripheral sensory neuron morphology and distribution, the mice were less susceptible to hypersensitivity to cold and mechanical stimuli after peripheral nerve injury. In contrast, thermal and mechanical nociceptive responses were similar to control mice in an inflammation-induced pain model. In the nerve injury model of neuropathic pain, WNK1/HSN2 contributed to a maladaptive decrease in the activity of the K(+)-Cl(-)cotransporter KCC2 by increasing its inhibitory phosphorylation at Thr(906)and Thr(1007), resulting in an associated loss of GABA (γ-aminobutyric acid)-mediated inhibition of spinal pain-transmitting nerves. Electrophysiological analysis showed that WNK1/HSN2 shifted the concentration of Cl(-)such that GABA signaling resulted in a less hyperpolarized state (increased neuronal activity) rather than a more hyperpolarized state (decreased neuronal activity) in mouse spinal nerves. Pharmacologically antagonizing WNK activity reduced cold allodynia and mechanical hyperalgesia, decreased KCC2 Thr(906)and Thr(1007)phosphorylation, and restored GABA-mediated inhibition (hyperpolarization) of injured spinal cord lamina II neurons. These data provide mechanistic insight into, and a compelling therapeutic target for treating, neuropathic pain after nerve injury.
authors.affiliation: Howard Hughes Medical Institute, Department of Neurobiology, Harvard Medical School, Boston, MA 02115, USA. Department of Neurosurgery, Boston Children's Hospital, Boston, MA 02124, USA. guy.rouleau@mcgill.ca kristopher.kahle@yale.edu.
authors.forename: Kristopher T
authors.initials: KT
authors.lastname: Kahle
authors.affiliation: Montreal Neurological Institute and Hospital, McGill University, Montréal, Quebec H3A 2B4, Canada. Department of Neurology and Neurosurgery, McGill University, Montréal, Quebec H3A 2B4, Canada.
authors.forename: Jean-François
authors.initials: JF
authors.lastname: Schmouth
authors.affiliation: Montreal Neurological Institute and Hospital, McGill University, Montréal, Quebec H3A 2B4, Canada. Department of Neurology and Neurosurgery, McGill University, Montréal, Quebec H3A 2B4, Canada.
authors.forename: Valérie
authors.initials: V
authors.lastname: Lavastre
authors.affiliation: F.M. Kirby Neurobiology Center, Boston Children's Hospital and Department of Neurobiology, Harvard Medical School, Boston, MA 02115, USA.
authors.forename: Alban
authors.initials: A
authors.lastname: Latremoliere
authors.affiliation: MRC Protein Phosphorylation and Ubiquitylation Unit, College of Life Sciences, University of Dundee, Dundee DD1 5EH, Scotland, UK.
authors.forename: Jinwei
authors.initials: J
authors.lastname: Zhang
authors.affiliation: F.M. Kirby Neurobiology Center, Boston Children's Hospital and Department of Neurobiology, Harvard Medical School, Boston, MA 02115, USA.
authors.forename: Nick
authors.initials: N
authors.lastname: Andrews
authors.affiliation: F.M. Kirby Neurobiology Center, Boston Children's Hospital and Department of Neurobiology, Harvard Medical School, Boston, MA 02115, USA.
authors.forename: Takao
authors.initials: T
authors.lastname: Omura
authors.affiliation: Montreal Neurological Institute and Hospital, McGill University, Montréal, Quebec H3A 2B4, Canada. Department of Neurology and Neurosurgery, McGill University, Montréal, Quebec H3A 2B4, Canada.
authors.forename: Janet
authors.initials: J
authors.lastname: Laganière
authors.affiliation: Montreal Neurological Institute and Hospital, McGill University, Montréal, Quebec H3A 2B4, Canada. Department of Neurology and Neurosurgery, McGill University, Montréal, Quebec H3A 2B4, Canada.
authors.forename: Daniel
authors.initials: D
authors.lastname: Rochefort
authors.affiliation: Montreal Neurological Institute and Hospital, McGill University, Montréal, Quebec H3A 2B4, Canada.
authors.forename: Pascale
authors.initials: P
authors.lastname: Hince
authors.affiliation: Montreal Neurological Institute and Hospital, McGill University, Montréal, Quebec H3A 2B4, Canada.
authors.forename: Geneviève
authors.initials: G
authors.lastname: Castonguay
authors.affiliation: Montreal Neurological Institute and Hospital, McGill University, Montréal, Quebec H3A 2B4, Canada.
authors.forename: Rébecca
authors.initials: R
authors.lastname: Gaudet
authors.affiliation: Department of Psychology and Centre for Research on Pain, McGill University, Montréal, Quebec H3A 0G1, Canada.
authors.forename: Josiane C S
authors.initials: JC
authors.lastname: Mapplebeck
authors.affiliation: Department of Psychology and Centre for Research on Pain, McGill University, Montréal, Quebec H3A 0G1, Canada.
authors.forename: Susana G
authors.initials: SG
authors.lastname: Sotocinal
authors.affiliation: Howard Hughes Medical Institute, Department of Neurobiology, Harvard Medical School, Boston, MA 02115, USA.
authors.forename: JingJing
authors.initials: J
authors.lastname: Duan
authors.affiliation: F.M. Kirby Neurobiology Center, Boston Children's Hospital and Department of Neurobiology, Harvard Medical School, Boston, MA 02115, USA.
authors.forename: Catherine
authors.initials: C
authors.lastname: Ward
authors.affiliation: Department of Neurosurgery, Boston Children's Hospital, Boston, MA 02124, USA.
authors.forename: Arjun R
authors.initials: AR
authors.lastname: Khanna
authors.affiliation: Department of Psychology and Centre for Research on Pain, McGill University, Montréal, Quebec H3A 0G1, Canada.
authors.forename: Jeffrey S
authors.initials: JS
authors.lastname: Mogil
authors.affiliation: Montreal Neurological Institute and Hospital, McGill University, Montréal, Quebec H3A 2B4, Canada. Department of Neurology and Neurosurgery, McGill University, Montréal, Quebec H3A 2B4, Canada.
authors.forename: Patrick A
authors.initials: PA
authors.lastname: Dion
authors.affiliation: F.M. Kirby Neurobiology Center, Boston Children's Hospital and Department of Neurobiology, Harvard Medical School, Boston, MA 02115, USA.
authors.forename: Clifford J
authors.initials: CJ
authors.lastname: Woolf
authors.affiliation: Institut des Neurosciences Cellulaires et Integratives, UPR 3212 Centre National de la Recherche Scientifique, and Universite de Strasbourg, 5 rue Blaise Pascal, F-67084 Strasbourg, France.
authors.forename: Perrine
authors.initials: P
authors.lastname: Inquimbert
authors.affiliation: Montreal Neurological Institute and Hospital, McGill University, Montréal, Quebec H3A 2B4, Canada. Department of Neurology and Neurosurgery, McGill University, Montréal, Quebec H3A 2B4, Canada. guy.rouleau@mcgill.ca kristopher.kahle@yale.edu.
authors.forename: Guy A
authors.initials: GA
authors.lastname: Rouleau
mesh_terms: D000818:Animals; D004195:Disease Models, Animal; D005091:Exons; D006930:Hyperalgesia / Q000235:genetics / Q000378:metabolism* / Q000503:physiopathology / Q000517:prevention & control; D051379:Mice; D008822:Mice, Transgenic; D015778:Minor Histocompatibility Antigens / Q000235:genetics / Q000378:metabolism; D009437:Neuralgia / Q000235:genetics / Q000378:metabolism* / Q000503:physiopathology / Q000517:prevention & control; D059348:Peripheral Nerve Injuries / Q000235:genetics / Q000378:metabolism / Q000503:physiopathology / Q000517:prevention & control; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D013127:Spinal Nerves / Q000378:metabolism* / Q000473:pathology; D027981:Symporters / Q000235:genetics / Q000378:metabolism; D009435:Synaptic Transmission; D000076204:WNK Lysine-Deficient Protein Kinase 1; D005680:gamma-Aminobutyric Acid / Q000235:genetics / Q000378:metabolism*; D000096922:K Cl- Cotransporters
chemical_list: D015778:Minor Histocompatibility Antigens; D017346:Protein Serine-Threonine Kinases; D027981:Symporters; D000076204:WNK Lysine-Deficient Protein Kinase 1; D005680:gamma-Aminobutyric Acid; D000096922:K Cl- Cotransporters; C498744:Wnk1 protein, mouse