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A non-functional galanin receptor-2 in a multiple sclerosis patient

The Pharmacogenomics Journal volume 19, pages 72–82 (2019)Cite this article

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Abstract

Multiple Sclerosis (MS) is an inflammatory neurodegenerative disease that affects approximately 2.5 million people globally. Even though the etiology of MS remains unknown, it is accepted that it involves a combination of genetic alterations and environmental factors. Here, after performing whole exome sequencing, we found a MS patient harboring a rare and homozygous single nucleotide variant (SNV; rs61745847) of the G-protein coupled receptor (GPCR) galanin-receptor 2 (GALR2) that alters an important amino acid in the TM6 molecular toggle switch region (W249L). Nuclear magnetic resonance imaging showed that the hypothalamus (an area rich in GALR2) of this patient exhibited an important volumetric reduction leading to an enlarged third ventricle. Ex vivo experiments with patient-derived blood cells (AKT phosphorylation), as well as studies in recombinant cell lines expressing the human GALR2 (calcium mobilization and NFAT mediated gene transcription), showed that galanin (GAL) was unable to stimulate cell signaling in cells expressing the variant GALR2 allele. Live cell confocal microscopy showed that the GALR2 mutant receptor was primarily localized to intracellular endosomes. We conclude that the W249L SNV is likely to abrogate GAL-mediated signaling through GALR2 due to the spontaneous internalization of this receptor in this patient. Although this homozygous SNV was rare in our MS cohort (1:262 cases), our findings raise the potential importance of impaired neuroregenerative pathways in the pathogenesis of MS, warrant future studies into the relevance of the GAL/GALR2 axis in MS and further suggest the activation of GALR2 as a potential therapeutic route for this disease.

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Acknowledgments

We acknowledge the support received from Associação Beneficente Alzira Denise Hertzog Silva (ABADHS), Mr. Waldemar Benassi, Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES, Brazil—Grants 23038.007775/2014-98; 7350-15-5 and 1197-79-4), Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq, Brazil—Grant: 480138/2013-3) and Fundação de Amparo à Pesquisa do Estado de São Paulo (Grants: 2013/24293-7, 2015/07925-5, 2016/06488-3). We thank Prof. Sérgio DJ Pena for the critical reading of this manuscript. ED-N and PSLO are research fellows from CNPq, Brazil. This work is dedicated to the memory of Mrs. Alice Benassi and Dr. Ricardo R. Brentani.

Author contributions

Genomics planning, data analysis, and project management: SG-R, DNN, and ED-N; Exome analysis, patient screening, and validations; SG-R, MGA; WHS; FOG-N; Specific clinical evaluations and diagnosis, sensitivity analysis, imaging data, EDSS graduations, sample collections, treatment, and follow-up of this and other RRMS patients: AAB, supported by ACS, CTG, DGB, VDM and RMC; Clinical evaluation, recruitment, and samples collection of other RRMS patients: SG-R, PPC, JANM; GSO, VBT, ASF, LMBS, CR, JCCL; Methylation analysis: HN and TSS; Planning and execution of ex vivo GALR2 signaling: RBS, WAS Jr, VV; Planning and execution of in vitro GALR2 experiments: MA, SJH, DBBT; Molecular modeling and molecular simulation: PSLO and JGCP; Bioinformatics: JESS, RV, and ITdS; Manuscript writing and overall project coordination: ED-N.

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Authors and Affiliations

  1. Lab. of Medical Genomics, AC Camargo Cancer Center, São Paulo, SP, Brazil

    Sheila Garcia-Rosa, Willem H Schreuder, Frederico O Gleber-Netto, Maria Amorim, Diana N Nunes & Emmanuel Dias-Neto

  2. Brazilian Biosciences National Laboratory (LNBio), Center for Research in Energy and Material (CNPEM), Campinas, SP, Brazil

    Daniela BB Trivella, José GC Pereira & Paulo SL Oliveira

  3. CAPES-University of Nottingham Programme in Drug Discovery, Queen’s Medical Centre University of Nottingham, Nottingham, NG7 2UH, United Kingdom

    Daniela BB Trivella & Maria Augusta Arruda

  4. Cell Signalling Research Group, School of Life Sciences, University of Nottingham, Nottingham, United Kingdom

    Daniela BB Trivella, Maria Augusta Arruda & Stephen J Hill

  5. Department of Neurosciences, Clinical Neuroimmunology Division, Medical School of Ribeirão Preto, University of São Paulo (USP), Ribeirão Preto, SP, Brazil

    Vanessa D Marques, Rodrigo M Conde, Antonio C Santos, Carlos T Guerreiro & Amilton A Barreira

  6. Department of Cellular and Molecular Biology, Medical School of Ribeirão Preto, University of São Paulo (USP), Ribeirão Preto, SP, Brazil

    Rodolfo B Serafim

  7. Department of Genetics, Ribeirão Preto Medical School, University of São Paulo (USP), Ribeirão Preto, SP, Brazil

    Julio CC Lorenzi, Greice A Molfetta, Thais S Sabedot, Houtan Noushmehr & Wilson A Silva Jr

  8. Curso de Pós-Graduação, Santa Casa de Belo Horizonte, Belo Horizonte, MG, Brazil

    Paulo P Christo & José A Nogueira-Machado

  9. Neurosciences Research Group, Faculdade de Ciências Médicas da Santa Casa de São Paulo, São Paulo, SP, Brazil

    Guilherme S Olival & Vania BT Marchitto

  10. Department of Neurology, Psychology and Psychiatry, School of Medicine of Botucatu, University of State of São Paulo (UNESP), Botucatu, SP, Brazil

    Doralina G Brum

  11. Neuroimmunology Unit, Institute of Biology, University of Campinas (UNICAMP), Campinas, Brazil

    Alessandro S Farias & Leonilda MB Santos

  12. Instituto Metrópole Digital, Federal University of Rio Grande do Norte, Natal, RN, Brazil

    Jorge ES Souza

  13. Lab. Virology (LIM52), Hospital das Clinicas HCFMUSP, Faculdade de Medicina, Universidade de Sao Paulo, São Paulo, SP, Brazil

    Camila M Romano

  14. Laboratory of Bioinformatics and Computational Biology, AC Camargo Cancer Center, Sao Paulo, Brazil

    Renan Valieris & Israel Tojal da Silva

  15. Center for Medical Genomics at HCFMRP, Universidade de Sao Paulo (USP), Ribeirão Preto, Brazil

    Wilson A Silva Jr

  16. School of Pharmaceutical Sciences, São Paulo State University (UNESP), Araraquara, SP, Brazil

    Valeria Valente

  17. Farmanguinhos, Fiocruz, Rio de Janeiro, Brazil

    Maria Augusta Arruda

  18. Centre of Membrane Proteins and Receptors (COMPARE), University of Birmingham and University of Nottingham, Nottingham, United Kingdom

    Maria Augusta Arruda & Stephen J Hill

  19. Lab. of Neurosciences (LIM27), Institute of Psychiatry, School of Medicine, University of São Paulo USP, São Paulo, SP, Brazil

    Emmanuel Dias-Neto

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  1. Sheila Garcia-Rosa
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Correspondence to Stephen J Hill, Amilton A Barreira or Emmanuel Dias-Neto.

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Garcia-Rosa, S., Trivella, D.B., Marques, V.D. et al. A non-functional galanin receptor-2 in a multiple sclerosis patient. Pharmacogenomics J 19, 72–82 (2019). https://doi.org/10.1038/s41397-018-0032-6

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