title: A Point Mutation in a lincRNA Upstream of GDNF Is Associated to a Canine Insensitivity to Pain: A Spontaneous Model for Human Sensory Neuropathies.
abstract: Human Hereditary Sensory Autonomic Neuropathies (HSANs) are characterized by insensitivity to pain, sometimes combined with self-mutilation. Strikingly, several sporting dog breeds are particularly affected by such neuropathies. Clinical signs appear in young puppies and consist of acral analgesia, with or without sudden intense licking, biting and severe self-mutilation of the feet, whereas proprioception, motor abilities and spinal reflexes remain intact. Through a Genome Wide Association Study (GWAS) with 24 affected and 30 unaffected sporting dogs using the Canine HD 170K SNP array (Illumina), we identified a 1.8 Mb homozygous locus on canine chromosome 4 (adj. p-val = 2.5x10-6). Targeted high-throughput sequencing of this locus in 4 affected and 4 unaffected dogs identified 478 variants. Only one variant perfectly segregated with the expected recessive inheritance in 300 sporting dogs of known clinical status, while it was never present in 900 unaffected dogs from 130 other breeds. This variant, located 90 kb upstream of the GDNF gene, a highly relevant neurotrophic factor candidate gene, lies in a long intergenic non-coding RNAs (lincRNA), GDNF-AS. Using human comparative genomic analysis, we observed that the canine variant maps onto an enhancer element. Quantitative RT-PCR of dorsal root ganglia RNAs of affected dogs showed a significant decrease of both GDNF mRNA and GDNF-AS expression levels (respectively 60% and 80%), as compared to unaffected dogs. We thus performed gel shift assays (EMSA) that reveal that the canine variant significantly alters the binding of regulatory elements. Altogether, these results allowed the identification in dogs of GDNF as a relevant candidate for human HSAN and insensitivity to pain, but also shed light on the regulation of GDNF transcription. Finally, such results allow proposing these sporting dog breeds as natural models for clinical trials with a double benefit for human and veterinary medicine.
authors.affiliation: CNRS, UMR 6290, Institut de Génétique et Développement de Rennes, Rennes, France.|Université Rennes 1, UEB, Biosit, Faculté de Médecine, Rennes, France.
authors.forename: Jocelyn
authors.initials: J
authors.lastname: Plassais
authors.affiliation: CNRS, UMR 6290, Institut de Génétique et Développement de Rennes, Rennes, France.|Université Rennes 1, UEB, Biosit, Faculté de Médecine, Rennes, France.
authors.forename: Laetitia
authors.initials: L
authors.lastname: Lagoutte
authors.affiliation: CNRS, UMR 6290, Institut de Génétique et Développement de Rennes, Rennes, France.|Université Rennes 1, UEB, Biosit, Faculté de Médecine, Rennes, France.
authors.forename: Solenne
authors.initials: S
authors.lastname: Correard
authors.affiliation: Department of Clinical Sciences, Faculté de Médecine Vétérinaire, University of Montreal, Montreal, Québec, Canada.
authors.forename: Manon
authors.initials: M
authors.lastname: Paradis
authors.affiliation: Clinique Vétérinaire Saint Bernard, Lomme, France.
authors.forename: Eric
authors.initials: E
authors.lastname: Guaguère
authors.affiliation: CNRS, UMR 6290, Institut de Génétique et Développement de Rennes, Rennes, France.|Université Rennes 1, UEB, Biosit, Faculté de Médecine, Rennes, France.
authors.forename: Benoit
authors.initials: B
authors.lastname: Hédan
authors.affiliation: CNRS, UMR 6290, Institut de Génétique et Développement de Rennes, Rennes, France.|Université Rennes 1, UEB, Biosit, Faculté de Médecine, Rennes, France.
authors.forename: Alix
authors.identifier: http://orcid.org/0000-0003-0588-8099
authors.initials: A
authors.lastname: Pommier
authors.affiliation: CNRS, UMR 6290, Institut de Génétique et Développement de Rennes, Rennes, France.|Université Rennes 1, UEB, Biosit, Faculté de Médecine, Rennes, France.
authors.forename: Nadine
authors.initials: N
authors.lastname: Botherel
authors.affiliation: UDEAR, Université de Toulouse, Ecole Nationale Vétérinaire de Toulouse, INSERM, Toulouse, France.
authors.forename: Marie-Christine
authors.identifier: http://orcid.org/0000-0001-6909-0153
authors.initials: MC
authors.lastname: Cadiergues
authors.affiliation: Clinique Vétérinaire de Saint-Cyr, Rennes, France.
authors.forename: Philippe
authors.initials: P
authors.lastname: Pilorge
authors.affiliation: Department of Clinical Sciences, Faculté de Médecine Vétérinaire, University of Montreal, Montreal, Québec, Canada.
authors.forename: David
authors.initials: D
authors.lastname: Silversides
authors.affiliation: CNRS, UMR 6290, Institut de Génétique et Développement de Rennes, Rennes, France.|Université Rennes 1, UEB, Biosit, Faculté de Médecine, Rennes, France.
authors.forename: Maud
authors.initials: M
authors.lastname: Bizot
authors.affiliation: Department of Biochemistry and Molecular Medicine, CHU Sainte-Justine, University of Montreal, Montreal, Québec, Canada.
authors.forename: Mark
authors.initials: M
authors.lastname: Samuels
authors.affiliation: Centre for Genomic Regulation (CRG), Barcelona, Spain.|Universitat Pompeu Fabra (UPF), Barcelona, Spain.|Institut Hospital del Mar d'Investigations Mèdiques (IMIM), Barcelona, Spain.
authors.forename: Carme
authors.identifier: http://orcid.org/0000-0002-7431-2088
authors.initials: C
authors.lastname: Arnan
authors.affiliation: Centre for Genomic Regulation (CRG), Barcelona, Spain.|Universitat Pompeu Fabra (UPF), Barcelona, Spain.|Institut Hospital del Mar d'Investigations Mèdiques (IMIM), Barcelona, Spain.
authors.forename: Rory
authors.identifier: http://orcid.org/0000-0003-4607-2782
authors.initials: R
authors.lastname: Johnson
authors.affiliation: CNRS, UMR 6290, Institut de Génétique et Développement de Rennes, Rennes, France.|Université Rennes 1, UEB, Biosit, Faculté de Médecine, Rennes, France.
authors.forename: Christophe
authors.initials: C
authors.lastname: Hitte
authors.affiliation: CNRS, UMR 6290, Institut de Génétique et Développement de Rennes, Rennes, France.|Université Rennes 1, UEB, Biosit, Faculté de Médecine, Rennes, France.
authors.forename: Gilles
authors.initials: G
authors.lastname: Salbert
authors.affiliation: CNRS, UMR 6290, Institut de Génétique et Développement de Rennes, Rennes, France.|Université Rennes 1, UEB, Biosit, Faculté de Médecine, Rennes, France.
authors.forename: Agnès
authors.initials: A
authors.lastname: Méreau
authors.affiliation: CNRS, UMR 6290, Institut de Génétique et Développement de Rennes, Rennes, France.|Université Rennes 1, UEB, Biosit, Faculté de Médecine, Rennes, France.
authors.forename: Pascale
authors.initials: P
authors.lastname: Quignon
authors.affiliation: CNRS, UMR 6290, Institut de Génétique et Développement de Rennes, Rennes, France.|Université Rennes 1, UEB, Biosit, Faculté de Médecine, Rennes, France.
authors.forename: Thomas
authors.initials: T
authors.lastname: Derrien
authors.affiliation: CNRS, UMR 6290, Institut de Génétique et Développement de Rennes, Rennes, France.|Université Rennes 1, UEB, Biosit, Faculté de Médecine, Rennes, France.
authors.forename: Catherine
authors.initials: C
authors.lastname: André
mesh_terms: D000818:Animals; D002874:Chromosome Mapping; D004285:Dogs; D005786:Gene Expression Regulation; D055106:Genome-Wide Association Study; D051100:Glial Cell Line-Derived Neurotrophic Factor / Q000235:genetics*; D009477:Hereditary Sensory and Autonomic Neuropathies / Q000235:genetics* / Q000503:physiopathology; D006801:Humans; D010146:Pain / Q000235:genetics* / Q000503:physiopathology; D000699:Pain Insensitivity, Congenital / Q000235:genetics* / Q000503:physiopathology; D017354:Point Mutation; D020641:Polymorphism, Single Nucleotide; D062085:RNA, Long Noncoding / Q000235:genetics*
chemical_list: D051100:Glial Cell Line-Derived Neurotrophic Factor; D062085:RNA, Long Noncoding