Genome-wide analysis identified novel susceptible genes of restless legs syndrome in migraineurs

• Published in: Journal of headache and pain Vol. 23; no. 1; p. 39
• Main Authors: Jiang, Yun-Jin, Fann, Cathy Shen-Jang, Fuh, Jong-Ling, Chung, Ming-Yi, Huang, Hui-Ying, Chu, Kuo-Chang, Wang, Yen-Feng, Hsu, Chia-Lin, Kao, Lung-Sen, Chen, Shih-Pin, Wang, Shuu-Jiun
• Format: Journal Article
• Language: English
• Published: Milan Springer Milan 29.03.2022; Springer Nature B.V; BMC
• Subjects: Amacrine cells; Animals; CRISPR; Danio rerio; Dopamine receptors; Embryos; Genes; Genetic diversity; Genome-wide association studies; Genome-Wide Association Study; Genomes; Headache; Humans; Internal Medicine; Leg; Medicine; Medicine & Public Health; Migraine; Neurology; Pain Medicine; Phenotypes; Research Article; Restless legs syndrome; Restless Legs Syndrome - complications; Restless Legs Syndrome - genetics; Sleep deprivation; Transcription; Translation; VSTM2L and CCDC141; Zebrafish; Zebrafish - genetics; Restless legs syndrome; Zebrafish; Genome-wide association study; Migraine; and; VSTM2L and CCDC141
• ISSN: 1129-2369; 1129-2377
• DOI: 10.1186/s10194-022-01409-9

Background Restless legs syndrome is a highly prevalent comorbidity of migraine; however, its genetic contributions remain unclear. Objectives To identify the genetic variants of restless legs syndrome in migraineurs and to investigate their potential pathogenic roles. Methods We conducted a two-stage genome-wide association study (GWAS) to identify susceptible genes for restless legs syndrome in 1,647 patients with migraine, including 264 with and 1,383 without restless legs syndrome, and also validated the association of lead variants in normal controls unaffected with restless legs syndrome ( n  = 1,053). We used morpholino translational knockdown (morphants), CRISPR/dCas9 transcriptional knockdown, transient CRISPR/Cas9 knockout (crispants) and gene rescue in one-cell stage embryos of zebrafish to study the function of the identified genes. Results We identified two novel susceptibility loci rs6021854 (in VSTM2L ) and rs79823654 (in CCDC141 ) to be associated with restless legs syndrome in migraineurs, which remained significant when compared to normal controls. Two different morpholinos targeting vstm2l and ccdc141 in zebrafish demonstrated behavioural and cytochemical phenotypes relevant to restless legs syndrome, including hyperkinetic movements of pectoral fins and decreased number in dopaminergic amacrine cells. These phenotypes could be partially reversed with gene rescue, suggesting the specificity of translational knockdown. Transcriptional CRISPR/dCas9 knockdown and transient CRISPR/Cas9 knockout of vstm2l and ccdc141 replicated the findings observed in translationally knocked-down morphants. Conclusions Our GWAS and functional analysis suggest VSTM2L and CCDC141 are highly relevant to the pathogenesis of restless legs syndrome in migraineurs.