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

Abstract

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.

In Search of a Target Gene for a Desirable Phenotype in Aquaculture: Genome Editing of Cyprinidae and Salmonidae Species

“…Genome editing has been used to model human tumors such as liver cancer [ 70 ], paraganglioma [ 71 ], skin melanoma [ 72 ], and epithelioid sarcoma [ 73 ]. Among neurologic diseases, amyotrophic lateral sclerosis [ 74 ], epilepsy [ 75 , 76 , 77 , 78 , 79 ], Hirschprung’s disease [ 80 ], autism [ 81 , 82 , 83 , 84 ], spastic paraplegia [ 85 ], restless legs syndrome [ 86 ], insomnia [ 87 ], microcephaly [ 46 ], neurotransmitter function of dopamine [ 88 ], monoamine [ 89 ], and others [ 90 , 91 ]. Cleft lip and cleft palate mutants also have been tested [ 92 ].…”

Genetics of migraine: where are we now?

“…With regard to genetic studies of associations and correlations, several large studies based on the comparison of GWAS data have shown the existence of a shared genetic susceptibility between migraine and various disorders, including psychiatric disorders [ 140 , 141 ], ischemic stroke [ 142 ], coronary artery disease [ 143 ], hypertension [ 144 , 145 ], sleep disorders [ 146 ], and also endometriosis [ 147 ], fibromuscular dysplasia [ 148 ], type 2 diabetes, hyperlipidemia, autoimmune diseases, asthma, other respiratory conditions [ 144 ], restless legs syndrome [ 149 ] and hemostatic profile [ 150 ]. In contrast, one study found no causal link between genetic susceptibility to migraine and Alzheimer’s disease, intelligence, and brain size [ 151 ].…”

Exploration of restless legs syndrome under the new concept: A review

“…Some reports appear to follow a non-Mendelian pattern of inheritance because, despite the apparent autosomal dominant mode of inheritance, the proportion of those affected (>50%) is higher than would be expected from typical autosomal dominant inheritance. Zimprich [ 3 ] proposed the possible role of genetic factors, trying to explain the characteristics of non-Mendelian inheritance patterns in RLS genetics and the existence of epigenetic inheritance in this disease family.…”

“…Many studies have attempted to identify the genes involved in the basic molecular mechanisms of this disease, but no single cause of RLS has been identified. [ 3 ] To date, genome-wide association studies of RLS have identified 13 different genes in several communities in Northern Europe, These genes are represented by chromosome 6p21.2 (BTBD9), 2p14 (MEIS1), 9p24.1-p23 (PTPRD), 15q23 (MAP2K5/SKOR1), and 16q12.1 (TOX3/BC034767) mononucleotides. [ 16 , 17 ] In addition, recent studies have identified 7 additional major susceptibility loci for RLS: RLS1 at 12q12-q21, RLS2 at 14q13-q21, RLS3 at 9p24-p22, RLS4 at 2q33, RLS5 at 20p13, RLS6 at 19p13, and RLS7 at 16p12.1.…”

“…The diagnostic features of RLS have been further explained and clarified [ 2 ] and have recently been updated. [ 2 ] Although other groups, such as the American Psychological Association, have established similar standards, [ 3 ] the definition of the IRLSSG group is still the most widely accepted and is therefore the main standard covered by this review. In addition, in order to maintain consistency and avoid confusion, despite previous efforts to propose a new, less stigmatizing medical term for RLS to be replaced by a new term for WED disease, this label is less known and has failed to gain widespread use and acceptance in recent years.…”