medRxiv preprint doi: https://doi.org/10.1101/2020.11.30.20241000; this version posted December 3, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license . Genome-wide association study identifies RNF123 locus as associated with chronic widespread musculoskeletal pain Md Shafiqur Rahman1, Bendik S Winsvold2,3, S.O. Chavez Chavez11, Sigrid Børte4,5,6, Yakov A. Tsepilov12,13, Sodbo Zh. Shapov12,14, HUNT All-In Pain (see below), Yurii Aulchenko12,13, Knut Hagen7,8, Egil A. Fors9, Kristian Hveem4,10, John-Anker Zwart2,4,6, J.B.J. van Meurs11, Maxim B. Freidin1, Frances M.K. Williams1* Affiliations: 1 Department of Twin Research and Genetic Epidemiology, School of Life Course Sciences, King's College London, London, United Kingdom 2 Department of Research, Innovation and Education, Division of Clinical Neuroscience, Oslo University Hospital, Oslo, Norway. 3 Department of Neurology, Oslo University Hospital, Oslo, Norway. 4 K. G. Jebsen Center for Genetic Epidemiology, Department of Public Health and Nursing, Faculty of Medicine and Health Sciences, Norwegian University of Science and Technology, Trondheim, Norway. 5 Research and Communication Unit for Musculoskeletal Health (FORMI), Department of Research, Innovation and Education, Division of Clinical Neuroscience, Oslo University Hospital, Oslo, Norway. 6 Institute of Clinical Medicine, Faculty of Medicine, University of Oslo, Oslo, Norway. 7 Department of Neuromedicine and Movement Science, Faculty of Medicine and Health Sciences, Norwegian University of Science and Technology, Trondheim, Norway. 8 Clinical Research Unit Central Norway, St. Olavs University Hospital, Trondheim, Norway. 9 Department of Public Health and Nursing, Faculty of Medicine and Health Sciences, Norwegian University of Science and Technology, Trondheim, Norway. 10 HUNT Research Center, Department of Public Health and Nursing, Faculty of Medicine and Health Sciences, Norwegian University of Science and Technology, Trondheim, Norway. 11 Department of Internal Medicine, Erasmus Medical Center, Rotterdam, The Netherlands. 12 Laboratory of Theoretical and Applied Functional Genomics, Novosibirsk State University, Novosibirsk, Russia. 13 PolyOmica, ‘s-Hertogenbosch, 5237 PA, the Netherlands 14 Laboratory of Recombination and Segregation Analysis, Institute of Cytology and Genetics, 10 Lavrentiev Avenue, Novosibirsk, 630090, Russia NOTE: This preprint reports new research that has not been certified by peer review and should not be used to guide clinical practice. 1 medRxiv preprint doi: https://doi.org/10.1101/2020.11.30.20241000; this version posted December 3, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license . *Corresponding Author Frances MK Williams PhD FRCP(E) Professor of Genomic Epidemiology & Hon Consultant Rheumatologist Department of Twin Research and Genetic Epidemiology, King's College London, Westminster Bridge Road, London SE1 7EH Phone: 0207188 5875 Email: [email protected] Banner "HUNT All-In Pain" Amy E Martinsen1,2,3, Anne Heidi Skogholt1,4, Ben Brumpton1, Cristen Willer5, Ingrid Heuch2, Ingunn Mundal6, Jonas Bille Nielsen1,5,7, Kjersti Storheim8,9, Kristian Bernhard Nilsen10,11, Lars Fritsche12, Laurent F. Thomas1,4,13,14, Linda M Pedersen2, Maiken E Gabrielsen1, Marianne Bakke Johnsen1,3,8, Marie Udnesseter Lie3,8, Oddgeir Holmen15, Synne Øien Stensland8,16, Wei Zhou17,18 Affiliations: 1 K. G. Jebsen Center for Genetic Epidemiology, Department of Public Health and Nursing, Faculty of Medicine and Health Sciences, Norwegian University of Science and Technology, Trondheim, Norway. 2 Department of Research, Innovation and Education, Division of Clinical Neuroscience, Oslo University Hospital, Oslo, Norway. 3 Institute of Clinical Medicine, Faculty of Medicine, University of Oslo, Oslo, Norway. 4 Department of Clinical and Molecular Medicine, Norwegian University of Science and Technology, Trondheim, Norway. 5 Department of Internal Medicine, Division of Cardiovascular Medicine, University of Michigan, Ann Arbor, 48109, MI, USA. 6 Department of Health Science, Molde University College, Molde, Norway. 7 Department of Epidemiology Research, Statens Serum Institut, Copenhagen, Denmark. 8 Research and Communication Unit for Musculoskeletal Health (FORMI), Department of Research, Innovation and Education, Division of Clinical Neuroscience, Oslo University Hospital, Oslo, Norway. 2 medRxiv preprint doi: https://doi.org/10.1101/2020.11.30.20241000; this version posted December 3, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license . 9 Faculty of Health Sciences, Department of physiotherapy, Oslo Metropolitan University, Oslo, Norway. 10 Department of Neuromedicine and Movement Science, Faculty of Medicine and Health Sciences, Norwegian University of Science and Technology (NTNU), Trondheim, Norway. 11 Department of Neurology, Oslo University Hospital, Oslo, Norway. 12 Center for Statistical Genetics, Department of Biostatistics, University of Michigan, Ann Arbor, 48109, MI, USA. 13 BioCore - Bioinformatics Core Facility, Norwegian University of Science and Technology, Trondheim. Norway. 14 Clinic of Laboratory Medicine, St.Olavs Hospital, Trondheim University Hospital, Trondheim, Norway. 15 HUNT Research Center, Department of Public Health and Nursing, Faculty of Medicine and Health Sciences, Norwegian University of Science and Technology, Trondheim, Norway. 16 NKVTS, Norwegian Centre for Violence and Traumatic Stress Studies. 17 Department of Computational Medicine and Bioinformatics, University of Michigan, Ann Arbor, MI, USA. 18 Analytic and Translational Genetics Unit, Massachusetts General Hospital, Boston, Massachusetts, USA. 3 medRxiv preprint doi: https://doi.org/10.1101/2020.11.30.20241000; this version posted December 3, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license . Abstract Background and Objectives: Chronic widespread musculoskeletal pain (CWP) is a symptom of fibromyalgia and a complex trait with poorly understood pathogenesis. CWP is heritable (48- 54%), but its genetic architecture is unknown and candidate gene studies have produced inconsistent results. We conducted a genome-wide association study to get insight into the genetic background of CWP. Methods: Northern Europeans from UK Biobank comprising 6,914 cases reporting pain all over the body lasting more than 3 months and 242,929 controls were studied. Replication of three lead genome-wide significant single nucleotide polymorphisms (SNPs) was attempted in 6 independent European cohorts (N=43,080; cases=14,177). Genetic correlations with risk factors, tissue specificity, and colocalization were examined. Results: Three genome-wide significant loci were identified (rs1491985, rs10490825, rs165599) residing within the genes RNF123, ATP2C1, and COMT. The RNF123 locus was replicated (meta-analysis p=0.0002), the ATP2C1 locus showed suggestive association (p=0.0227), and the COMT locus was not replicated. Partial genetic correlation between CWP and depressive symptoms, body mass index, age of first birth, and years of schooling were identified. Tissue specificity and colocalization analysis highlight the relevance of skeletal muscle in CWP. Conclusions: We report a novel association of RNF123 locus with CWP and suggest a role of ATP2C1, consistent with a role of calcium regulation in CWP. The association to COMT, one of the most studied genes in chronic pain field, was not confirmed in the replication analysis. Keywords: Chronic widespread pain, fibromyalgia, genome-wide association study, twin, RNF123. 4 medRxiv preprint doi: https://doi.org/10.1101/2020.11.30.20241000; this version posted December 3, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license . Key messages What is already known about this subject? • Chronic widespread musculoskeletal pain (CWP) is a primary diagnostic feature of fibromyalgia. • CWP is moderately heritable, but precise genes involved in the pathogenesis of CWP are yet to be identified. What does this study add? • This is the largest genetic study conducted on CWP to date and identified novel genetic risk loci (RNF123 and ATP2C1). • The genetic signal points to peripheral pain mechanisms in CWP, and shows genetic correlation with other traits, including BMI and depression. How might this impact on clinical practice or future developments? • The findings add to etiological basis of CWP. 5 medRxiv preprint doi: https://doi.org/10.1101/2020.11.30.20241000; this version posted December 3, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted medRxiv a license to