Abstract
Chronic sleep disturbances, associated with cardiometabolic diseases, psychiatric disorders and all-cause mortality1,2, affect 25–30% of adults worldwide3. Although environmental factors contribute substantially to self-reported habitual sleep duration and disruption, these traits are heritable4,5,6,7,8,9 and identification of the genes involved should improve understanding of sleep, mechanisms linking sleep to disease and development of new therapies. We report single- and multiple-trait genome-wide association analyses of self-reported sleep duration, insomnia symptoms and excessive daytime sleepiness in the UK Biobank (n = 112,586). We discover loci associated with insomnia symptoms (near MEIS1, TMEM132E, CYCL1 and TGFBI in females and WDR27 in males), excessive daytime sleepiness (near AR–OPHN1) and a composite sleep trait (near PATJ (INADL) and HCRTR2) and replicate a locus associated with sleep duration (at PAX8). We also observe genetic correlation between longer sleep duration and schizophrenia risk (rg = 0.29, P = 1.90 × 10−13) and between increased levels of excessive daytime sleepiness and increased measures for adiposity traits (body mass index (BMI): rg = 0.20, P = 3.12 × 10−9; waist circumference: rg = 0.20, P = 2.12 × 10−7).
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Main
Rather than being 'secondary', disordered sleep may have an important role in the etiology and maintenance of physical and mental health1,2. Heritability has been estimated at ∼40% for sleep duration4,6,7,8, 25–45% for insomnia9 and 17% for excessive daytime sleepiness9, but few genetic factors are known. A Mendelian mutation in BHLHE41 (encoding p.Pro385Arg) associated with short sleep duration has been identified and was confirmed in mouse models10. Genome-wide association studies (GWAS) for sleep duration have been reported11,12,13,14, but only an association at the PAX8 locus reached genome-wide significance and was confirmed across ancestry groups12. There are several reported loci associated with restless legs syndrome (RLS) and narcolepsy, but no robust genetic loci have been associated with insomnia symptoms or excessive daytime sleepiness15,16.
We and others have performed a GWAS for chronotype in the UK Biobank17,18 and a 23andMe participant sample19. To identify genetic variants that contribute to self-reported sleep duration, insomnia symptoms and excessive daytime sleepiness and link them with other conditions, we performed GWAS using phenotype measures in UK Biobank participants of European ancestry. Variation in sleep duration, insomnia symptoms and excessive daytime sleepiness was significantly associated with age, sex, principal components of ancestry, genotyping array, depression, psychiatric medication use, self-reported sleep apnea and BMI (Supplementary Table 1), as previously reported20,21,22,23. Together, age, sex and principal components explained 0.4%, 3.0% and 1.3% of variation in sleep duration, insomnia symptoms and excessive daytime sleepiness, respectively. Strong and significant pairwise phenotypic correlation was seen between the traits overall and within each sex, with limited correlation observed with chronotype (Fig. 1 and Supplementary Fig. 1).
We performed GWAS analyses of sleep duration, insomnia symptoms and excessive daytime sleepiness using linear/logistic regression adjusting for age, sex, ten principal components and genotyping array. We identified 9 genome-wide significant (P < 5 × 10−8) and 14 suggestive (P < 5 × 10−7 to P = 5 × 10−8) loci (Fig. 2, Table 1 and Supplementary Figs. 2 and 3). For sleep duration (n = 111,975), the strongest association was observed at the PAX8 locus (rs62158211[T]: β (s.e.) = 2.34 (0.30) min/allele, P = 4.7 × 10−14, effect allele frequency (EAF) = 0.213; Fig. 2a), confirming a previously reported association (r2 = 0.96, D′ = 1 to lead SNP rs1823125 in 1000 Genomes Project CEU (European-ancestry) individuals)12. For insomnia symptoms (n = 32,155 cases and 26,973 controls), we observed significant associations within MEIS1 (rs113851554[T]: odds ratio (OR) (95% confidence interval (CI)) = 1.26 (1.20–1.33), P = 9.1 × 10−19, EAF = 0.057; Fig. 2b), a homeobox gene implicated in motor neuron connectivity in Drosophila melanogaster24,25, retinal and lens development in mouse26 and substance P expression in the amygdala in humans27; near TMEM132E (rs145258459[C]: OR (95% CI) = 1.23 (1.13–1.35), P = 2.1 × 10−8, EAF = 0.983; Fig. 2c), which belongs to a gene family whose members have roles in brain development28, panic/anxiety29 and bipolar disorder30, suggesting a link between insomnia symptoms and an underlying broader sensitivity to anxiety and stress; and near CYCL1 (rs5922858[G]: OR (95% CI) = 1.12 (1.07–1.16), P = 1.28 × 10−8, EAF = 0.849; Fig. 2d), a locus previously associated (P = 1 × 10−6) with alcohol dependence comorbid with depressive symptoms31. Sex-stratified analyses identified an additional female-specific signal near TGFBI (rs3792900[C]: OR (95% CI) = 1.10 (1.07–1.14), P = 2.16 × 10−8, EAF = 0.470; Table 1, Supplementary Fig. 3q,r and Supplementary Table 2), which encodes an extracellular matrix protein responsible for human corneal dystrophy32, and a male-specific signal near WDR27 (rs13192566[G]: OR (95% CI) = 1.14 (1.09–1.20), P = 3.2 × 10−8, EAF = 0.860; Table 1, Supplementary Figs. 3s,t and 4, and Supplementary Table 2), which encodes a scaffold protein. Independent associations at both loci are observed with type 1 diabetes, suggesting that these proteins may have a role in autoimmunity33,34,35. For excessive daytime sleepiness (n = 111,648), we identified a signal near the androgen receptor gene AR (rs73536079[T]: β (s.e.) = 0.634 (0.115), P = 3.94 × 10−8, EAF = 0.002; Fig. 2e), with no sex-specific effects. Secondary analyses after additional adjustment for depression or BMI identified a signal near ROBO1 (depression adjustment n = 107,440; rs182765975[T]: β (s.e.) = 0.099 (0.018), P = 3.33 × 10−8, EAF = 0.003; Table 1 and Supplementary Fig. 3o), which encodes a neuronal axon guidance receptor previously implicated in dyslexia36, and a signal near another member of the TMEM132 family, TMEM132B (BMI adjustment n = 75,480; rs142261172[A]: β (s.e.) = 0.106 (0.018), P = 9.06 × 10−9, EAF = 0.004; Table 1 and Supplementary Fig. 3p). Conditional analyses did not identify independent association signals (Supplementary Table 3). Sensitivity analyses adjusting for factors influencing sleep traits, including self-reported sleep apnea, depression, psychiatric medication use, smoking, socioeconomic status, employment status, marital status and snoring, did not significantly alter results for primary association signals (Supplementary Table 4).
The leading associations overlap interesting candidate genes whose expression is enriched in hypocretin-expressing neurons in mice and zebrafish37,38, that are differentially expressed in sleep-deprived rats39 and/or that regulate sleep in Drosophila40. Credible set analyses41 highlighted a number of potential causal variants at each locus (Table 1), and future experimental studies will be necessary to characterize the functions of these variants in regulating sleep traits. Bioinformatic annotations42 offer an initial opportunity for in silico functional interpretation of the variants (Supplementary Fig. 5 and Supplementary Table 5). For example, multiple variants for all three traits are predicted to disrupt binding of FOXP1, a neural transcriptional repressor implicated in intellectual disability, autism and language impairment43. Interestingly, the locus associated with sleep duration encompassing PAX8 is adjacent to the only chromosomal fusion site that arose since the divergence of humans from other hominids ∼5 million years ago44,45, and the new genomic structure created by this unique evolutionary history may have a causal role in sleep duration. Pathway analysis46 of significant and suggestive loci identified enrichment of genes associated with immune, neurodevelopmental, pituitary and communication disorders (P < 0.01) and genes enriched for binding sites for the stress-responsive transcription factor heat shock factor 1 (HSF1) and the endoplasmic reticulum stress- and unfolded-protein-responsive transcription factor HERPUD1 (Supplementary Tables 6 and 7).
Aside from the lead SNPs in the PAX8 region and a variant in the DRD2 region47 for sleep duration, we found limited evidence of association for previously published candidate gene or GWAS signals (Pmeta < 5 × 10−5; Supplementary Table 8) or for regions encompassing core clock genes (Supplementary Fig. 6). Our findings for the GWAS on sleep duration largely overlap with those of Jones et al.18, despite differences in exclusion criteria and analytical approach. In particular, our study excluded shift workers (n = 6,557), sleep medication users (n = 1,184) and first- to third-degree relatives (n = 7,980), whereas the linear mixed-model analyses by Jones et al.18 included these populations, leading to a larger sample size (n = 127,573). Likely because of this increased power, Jones et al.18 identified two additional signals at VRK2 that did not attain genome-wide significance in our study (rs1380703[A]: β (s.e.) = 1.50 (0.30) min/allele, P = 8.43 × 10−8 and rs17190618[T]: β (s.e.) = 1.60 (0.34) min/allele, P = 3.80 × 10−6).
Trait heritability calculated as the proportion of trait variance due to additive genetic factors measured here (observed-scale SNP heritability, h2 (s.e.)) was 10.3% (0.006%) for sleep duration, 20.6% (0.011%) for insomnia symptoms and 8.4% (0.006%) for daytime sleepiness (BOLT-REML variance-components analysis48). LD score regression analysis49 confirmed no residual population stratification (intercept (s.e.): sleep duration, 1.012 (0.008); insomnia symptoms, 1.003 (0.008); excessive daytime sleepiness, 1.005 (0.007)). Tests for enrichment of heritability by functional class using an LD score regression approach50 identified excess heritability across active transcriptional regions for insomnia symptoms and across genomic regions conserved in mammals for all three sleep traits. Consistent with these findings, heritability enrichment in conserved regions was seen for traits demonstrating significant genetic correlation with sleep (Fig. 3 and Supplementary Table 9).
Sleep duration, insomnia symptoms, excessive daytime sleepiness and chronotype are significantly correlated, both at the phenotype and genetic levels (Fig. 1), with greater pairwise correlations in males than in females (Supplementary Fig. 1). Thus, to find loci common to sleep traits, we performed a multiple-trait GWAS51. We identified two new association signals near HCRTR2 and PATJ (INADL) and found that PAX8 and MEIS1 associations influenced multiple sleep traits (Fig. 2, Table 2 and Supplementary Fig. 7). HCRTR2 encodes hypocretin receptor 2, the primary receptor of two receptors for wake-promoting orexin neuropeptides52 involved in narcolepsy and regulation of sleep. Notably, the minor allele at rs3122163 (C) showed subthreshold association with shorter sleep duration and morningness chronotype, suggesting gain of function, but no association with insomnia symptoms. Assessment of objective sleep measures and functional and physiological follow-up should yield important insights into orexin receptor signaling, a pathway important for the pharmacological treatment of narcolepsy53 and insomnia54. PATJ encodes a membrane protein involved in the formation of tight junctions and is implicated in photoreception in mice and Drosophila55,56. The INADL protein is reported to interact with HTR2A (ref. 57), a serotonin receptor with a known role in sleep regulation58,59.
Our strongest association for insomnia symptoms fell within MEIS1, a locus previously associated with RLS in GWAS60. Our lead SNP, rs113851554, and the correlated 3′ UTR variant rs11693221 (pairwise r2 = 0.69, D′ = 0.90 in 1000 Genomes Project European (EUR) individuals) represent the strongest known genetic risk factor for RLS and were identified in MEIS1 sequencing studies61,62 following up the original RLS GWAS signal (rs2300478)60,63. Conditional analysis suggests that only one underlying signal, detected by the lead SNP rs113851554 in our GWAS, explains the association of all three SNPs with insomnia symptoms (Supplementary Fig. 8 and Supplementary Table 10). To further investigate the extent of overlap between RLS and insomnia symptoms, we tested a weighted genetic risk score (GRS) for RLS64,65 and found that it was also associated with insomnia symptoms with concordant direction of allelic effects (OR (95% CI) = 1.06 (1.05–1.07) per RLS risk allele, P = 1.17 × 10−21; Supplementary Table 11). Weighting of the RLS GWAS alleles by SNP effect on periodic limb movements (PLMs) did not substantially alter the overall results (Supplementary Table 11). Interestingly, recent data indicating that thalamic glutamatergic activity is increased in RLS provide evidence of an underlying propensity for hyperarousal in RLS66, which is also a core feature of insomnia. Future analyses of pairwise bidirectional causal effects for all three traits will be necessary to determine whether shared genetic associations correspond to causality, partial mediation or pleiotropy.
Strong epidemiological associations of sleep duration, insomnia symptoms and daytime sleepiness have been observed with disease traits, but the extent to which the underlying genetics are shared is unknown. Therefore, we tested for genome-wide genetic correlation between findings from our sleep GWAS and those from publicly available GWAS for 20 phenotypes spanning a range of cognitive, neuropsychiatric, anthropometric, cardiometabolic and autoimmune traits using LD score regression67 (Fig. 4 and Supplementary Table 12).
Genetic correlations demonstrated a strong biological link between longer sleep duration and risk of schizophrenia (rg = 0.29, P = 1 × 10−13), as suggested by previous reports18,47,68. Furthermore, a schizophrenia GRS (96 variants) was associated with longer sleep duration (β (s.e.) = 1.44 (0.36) min/allele, P = 2.56 × 10−4) (interquartile range = 2.3 h), although a variety of sleep behaviors are seen in patients with schizophrenia69,70,71. Significant genetic correlation between low birth weight and longer sleep duration (rg = −0.27, P = 1 × 10−4) may reflect shared links between genetically determined insulin secretion or action pathways underlying fetal growth72,73 and long sleep duration. In support of this hypothesis, significant genetic correlation was observed by Jones et al.18 between oversleeping and both fasting insulin and risk of type 2 diabetes in UK Biobank. Genetic correlation between sleep duration and Crohn's disease risk (rg = 0.18, P = 1 × 10−3) is also consistent with epidemiological observations74.
Significant genetic correlation was also found between increased insomnia symptoms and major depression, adverse glycemic traits, increased adiposity and fewer years of education and between excessive daytime sleepiness and increased adiposity (all P < 1 × 10−3), further highlighting biological overlap of sleep traits with metabolic traits, psychiatric traits and educational attainment17. In support of this overlap, studies have shown that experimentally suppressing slow-wave sleep leads to decreased insulin sensitivity and impaired glucose tolerance75,76. Notably, a GRS for fasting insulin was not significantly associated with insomnia symptoms (7 SNPs, OR (95% CI) = 1.01 (0.99–1.03), P = 0.51). Finally, consistent with a well-established but poorly understood link between excessive daytime sleepiness and obesity77,78, a GRS for BMI was associated with excessive daytime sleepiness (95 SNPs, β (s.e.) = 0.002 (0.0004) sleepiness category/allele, P = 1.67 × 10−4) but not with insomnia symptoms (OR (95% CI) = 1.00 (0.998–1.003), P = 0.73).
Moving forward, replication and systematic testing of genetic correlations in larger samples will be needed. Notably, genetic correlation testing between insomnia and RLS should be examined but was not possible here because RLS consortium GWAS results were not available. Additionally, identifying causal relationships between genetically correlated traits may be difficult, and findings using Mendelian randomization approaches will need cautious interpretation given potential selection biases in UK Biobank79,80,81.
In summary, in a GWAS of sleep traits, we identified new genetic loci that point to previously unstudied variants that might modulate the hypocretin–orexin system and retinal development and influence genes expressed in the cerebral cortex. Furthermore, genome-wide analysis suggests that sleep traits have underlying genetic pathways in common with neuropsychiatric and metabolic diseases. This work should advance understanding of the molecular processes underlying sleep disturbances and open new avenues of treatment for sleep disorders and related disorders.
Methods
Population and study design.
Study participants were from the UK Biobank study, described in detail elsewhere80,81,82. In brief, the UK Biobank is a prospective study of >500,000 people living in the UK. All people in the National Health Service registry who were aged 40–69 years and living <25 miles from a study center were invited to participate from 2006–2010. In total, 503,325 participants were recruited from over 9.2 million mailed invitations. Self-reported baseline data were collected by questionnaire, and anthropometric assessments were performed. For the current analysis, individuals of non-white ethnicity were excluded to avoid confounding effects. All participants provided informed consent to the UK Biobank.
Sleep quality, sleep quantity and covariate measures.
Study subjects self-reported sleep duration, insomnia symptoms, excessive daytime sleepiness, depression, medication use, age, sex, height and weight on a touchscreen questionnaire. For sleep duration, subjects were asked, “About how many hours sleep do you get in every 24 hours? (please include naps),” with responses in hour increments. To assess insomnia symptoms, subjects were asked, “Do you have trouble falling asleep at night or do you wake up in the middle of the night?” with responses “never/rarely,” “sometimes,” “usually” and “prefer not to answer.” To assess daytime sleepiness, subjects were asked, “How likely are you to doze off or fall asleep during the daytime when you don't mean to? (e.g. when working, reading or driving),” with responses “never/rarely,” “sometimes,” “often,” “all the time,” “don't know” and “prefer not to answer.” Approximately 500,000 subjects answered these questions, but only the 120,286 unrelated individuals with genetic data and European ancestry were considered for analysis. Subjects with self-reported shift work (n = 6,557) or sleep medication use (n = 1,184) were excluded. Subjects who responded “do not know” or “prefer not to answer” were set to missing. Sleep duration and excessive daytime sleepiness were untransformed and treated as continuous variables, with daytime sleepiness coded as 1–4. The insomnia symptoms trait was dichotomized into controls (“never/rarely”) and cases (“usually”). Covariates used in sensitivity analyses included self-reported sleep apnea, BMI, depression, psychiatric medication use, socioeconomic, smoking, employment and marital status, and snoring, and secondary GWAS for sleepiness included adjustment for BMI or depression. Sleep apnea cases were defined on the basis of ICD-10 diagnosis code (391 cases). BMI at baseline visit was calculated from entries of height and weight (n = 75,540 with available data). Depression was reported in answer to the question “How often did you feel down, depressed or hopeless mood in last 2 weeks?” (cases (n = 4,242) answered “more than half the days” or “nearly every day”). Medication use was self-reported as part of the initial UK Biobank interview. Our list of psychiatric medications for sensitivity analysis included the four most widely used: fluoxetine (Prozac), citalopram (Cipranol), paroxetine (Seroxat) and sertraline (Lustral). Our list of sleep medications included the 21 most widely used sleep medications in the UK Biobank (oxazepam, meprobamate, medazepam, bromazepam, lorazepam, clobazam, chlormezanone, temazepam, nitrazepam, lormetazepam, diazepam, zopiclone, triclofos, methyprylone, prazepam, triazolam, ketazolam, dichloralphenazone, clomethiazole, zaleplon and butobarbital). Smoking status was self-reported as past and current smoking behavior, and individuals were classified into those with “current” or “past” smoking or “never” smokers. Socioeconomic status was represented by the Townsend deprivation index, on the basis of national census data obtained immediately preceding participation in UK Biobank. Employment status was self-reported (cases were retired; controls were currently employed). Marital status was derived from self-reported household occupancy and relatedness data. Snoring was reported in answer to the question “Does your partner or a close relative or friend complain about your snoring?”
Genotyping, quality control and imputation.
Of the ∼500,000 subjects with phenotype data in the UK Biobank, ∼153,000 are currently genotyped. Genotyping was performed by the UK Biobank, and genotyping, quality control and imputation procedures are described in detail at the UK Biobank website (http://biobank.ctsu.ox.ac.uk/). In brief, blood, saliva and urine were collected from participants, and DNA was extracted from buffy coat samples. Participant DNA was genotyped on two arrays, UK BiLEVE and UKB Axiom, with >95% common content. Genotypes were called using Affymetrix Power Tools software. Sample and SNP quality control were performed. Samples were removed for high rates of missingness or heterozygosity (480 samples), short runs of homozygosity (8 samples), relatedness (1,856 samples) and sex mismatches (191 samples). Genotypes for 152,736 samples passed sample quality control (∼99.9% of total samples). SNPs were excluded if they did not pass quality control filters across all 33 genotyping batches. Batch effects were identified through frequency and Hardy–Weinberg equilibrium tests (P < 1 × 10−12). Before imputation, 806,466 SNPs passed quality control in at least one batch (>99% of the array content). Population structure was captured by principal-component analysis on the samples using a subset of high-quality (missingness <1.5%), high-frequency (>2.5%) SNPs (∼100,000 SNPs) and identified the subsample of European descent. Imputation of autosomal SNPs was performed to a merged reference panel comprising the Phase 3 1000 Genomes Project and UK10K panels using IMPUTE2 (ref. 83). Data were prephased using SHAPEIT3 (ref. 84). In total, 73,355,677 SNPs, short indels and large structural variants were imputed. X-chromosome data were imputed separately, using Eagle 2.0 for prephasing with the --X chromosome flag (no reference panel) for the entire cohort85 and IMPUTE2 with the Phase 3 1000 Genomes Project reference panel for imputation with the --chrX flag on 500-kb chunks in randomly assigned subsets of 30,000 individuals. Post-imputation quality control was performed as previously outlined (http://biobank.ctsu.ox.ac.uk/), and an imputation INFO score cutoff of 0.8 was applied. For GWAS, we further excluded SNPs with minor allele frequency (MAF) <0.001, per-SNP missingness >10% and per-sample missingness >40%. In total, up to 112,586 samples of European descent with high-quality genotyping and complete phenotype/covariate data were used for these analyses.
Statistical analysis.
Phenotypic correlation analysis was performed with the Spearman test in R using the Hmisc package. Genetic association analysis for autosomes was performed in SNPTEST86,87 with the 'expected' method using an additive genetic model adjusted for age, sex, ten principal components and genotyping array. Genome-wide association analysis was performed separately for sleep duration, insomnia symptoms and excessive daytime sleepiness with a genome-wide significance threshold of 5 × 10−8 for each GWAS. We had 80% power to detect the following effects: sleep duration, β = 0.045 h (2.7 min); insomnia symptoms, OR = 1.07; excessive daytime sleepiness, β = 0.021 units (assuming MAF = 0.1, P = 5 × 10−7); we had 80% power to detect the following effects: sleep duration, β = 0.048 h (2.9 min); insomnia symptoms, OR = 1.08; excessive daytime sleepiness, β = 0.023 units (assuming MAF = 0.1, P = 5 × 10−8). X-chromosome analysis was performed in PLINK 1.9 (ref. 88) using linear/logistic regression with separate analysis of the pseudoautosomal regions using the split-chromosome flag, adjusting for sex, age, ten principal components and genotyping array. For the X-chromosome signal at rs73536079, we verified using principal-component analysis that all carriers of the minor allele fell within the major European-ancestry cluster. Follow-up analyses of genome-wide suggestive and significant loci in the primary analyses included covariate sensitivity analyses individually adjusting for sleep apnea, depression, psychiatric medication use, socioeconomic, smoking, employment and marital status, and snoring or BMI (on top of the baseline model adjusting for age, sex, ten principal components and genotyping array). Sensitivity analysis was conducted only in the subset of subjects with data for all secondary covariates (n = 75,477 for sleep duration, n = 39,812 for insomnia symptoms and n = 75,640 for excessive daytime sleepiness). Analysis of enrichment for disease-associated gene sets and transcription factors was performed in WebGestalt46 using the human genome as the reference set, Benjamini–Hochberg adjustment for multiple testing and a minimum of two genes per category. Sex-specific GWAS were performed in PLINK 1.9 using linear/logistic regression stratified by sex adjusting for age, ten principal components of ancestry and genotyping array. We used a hard-call genotype threshold of 0.1 (calls with greater than 0.1 were treated as missing), a SNP imputation quality threshold of 0.80 and a MAF threshold of 0.001. Regional association plots were generated with LocusZoom using the hg19 Nov2014 EUR reference panel to determine background linkage disequilibrium89.
Trait heritability was calculated as the proportion of trait variance due to additive genetic factors across the autosomes, measured in this study using BOLT-REML48 to leverage the power of raw genotype data together with data for low-frequency variants (MAF ≥ 0.001). For multiple-trait genome-wide association analysis, we applied the CPASSOC package developed by Zhu et al.51 to combine association evidence for chronotype, sleep duration, insomnia symptoms and excessive daytime sleepiness. CPASSOC provides two statistics, SHom and SHet. SHom is similar to statistics generated by the fixed-effects meta-analysis method90 but accounts for the correlation of summary statistics due to trait correlation. SHom uses the sample size for a trait as a weight instead of variance, so that it is possible to combine traits with different measurement scales. SHet is an extension of SHom, but power can be improved when the genetic effect sizes are different for different traits. The distribution of SHet values under the null hypothesis of no association was obtained through an estimated beta distribution. To calculate the SHom and Shet statistics, a correlation matrix is required to account for the correlation among traits or resulting from overlapping or related samples from different cohorts. In this study, we directly provide the correlation matrix calculated from the residuals of four sleep traits after adjusting for age, sex, principal components of ancestry and genotyping array. Post-GWAS genome-wide genetic correlation analysis by LD score regression (LDSC)67 was conducted using all UK Biobank SNPs also found in HapMap 3 (ref. 89) and included publicly available data from 20 published GWAS, using a significance threshold of P = 0.0026 after Bonferroni correction for all 20 tests performed. As expected, the observed heritability estimates from LDSC67 using summary statistics for HapMap 3 were lower (5.7% (0.0065%) for sleep duration, 13.3% (0.0123%) for insomnia symptoms and 5.3% (0.005%) for sleepiness) than those calculated by BOLT-REML48 using primary data (10.3% (0.006%) for sleep duration, 20.6% (0.011%) for insomnia symptoms and 8.4% (0.006%) for sleepiness) because the HapMap 3 panel was restricted to variants with MAF >5%. LDSC estimates genetic correlation between two traits (ranging from −1 to 1) from summary statistics using the facts that the GWAS effect size estimate for each SNP incorporates the effects of all SNPs in linkage disequilibrium with that SNP, SNPs with high linkage disequilibrium have higher χ2 statistics than SNPs with low linkage disequilibrium, and a similar relationship is observed when single-study test statistics are replaced with the product of the z scores from two studies of traits with some correlation67. Furthermore, genetic correlation is possible between case–control studies and studies of quantitative traits, as well as within these study types. We performed a weighted GRS analysis using risk scores for RLS, schizophrenia, BMI and fasting insulin. Risk score SNPs passed the genome-wide significance threshold (P < 5 × 10−8) in recent large-scale GWAS and were present in the UK Biobank (RLS, 7 SNPs65, Supplementary Table 11; schizophrenia, 96 SNPs91; BMI, 95 SNPs92; fasting insulin, 7 SNPs93). Independent SNPs were identified, and β estimates were recorded for calculation of the weighted risk score. The GRS was calculated by summing the products of the risk allele count multiplied by the effect reported in the discovery GWAS. The additive genotype model was used for all SNPs. We performed partitioning of heritability using the 25 precomputed functional annotations available through LDSC, which were curated from large-scale robust data sets50. Enrichment both in functional regions and in an expanded region (+500 bp) around loci corresponding to each functional class was calculated to prevent the estimates from being biased upward by enrichment in nearby regions. The multiple-testing threshold was determined using conservative Bonferroni correction (P = 0.05/25 classes).
Data availability.
Summary GWAS statistics will be made available at the UK Biobank website (http://biobank.ctsu.ox.ac.uk/).
References
Fernandez-Mendoza, J. & Vgontzas, A.N. Insomnia and its impact on physical and mental health. Curr. Psychiatry Rep. 15, 418 (2013).
Luyster, F.S., Strollo, P.J. Jr., Zee, P.C. & Walsh, J.K. Sleep: a health imperative. Sleep 35, 727–734 (2012).
Stranges, S., Tigbe, W., Gómez-Olivé, F.X., Thorogood, M. & Kandala, N.B. Sleep problems: an emerging global epidemic? Findings from the INDEPTH WHO-SAGE study among more than 40,000 older adults from 8 countries across Africa and Asia. Sleep 35, 1173–1181 (2012).
de Castro, J.M. The influence of heredity on self-reported sleep patterns in free-living humans. Physiol. Behav. 76, 479–486 (2002).
Evans, D.S. et al. Habitual sleep/wake patterns in the Old Order Amish: heritability and association with non-genetic factors. Sleep 34, 661–669 (2011).
Heath, A.C., Eaves, L.J., Kirk, K.M. & Martin, N.G. Effects of lifestyle, personality, symptoms of anxiety and depression, and genetic predisposition on subjective sleep disturbance and sleep pattern. Twin Res. 1, 176–188 (1998).
Heath, A.C., Kendler, K.S., Eaves, L.J. & Martin, N.G. Evidence for genetic influences on sleep disturbance and sleep pattern in twins. Sleep 13, 318–335 (1990).
Partinen, M., Kaprio, J., Koskenvuo, M., Putkonen, P. & Langinvainio, H. Genetic and environmental determination of human sleep. Sleep 6, 179–185 (1983).
Wing, Y.K. et al. Familial aggregation and heritability of insomnia in a community-based study. Sleep Med. 13, 985–990 (2012).
He, Y. et al. The transcriptional repressor DEC2 regulates sleep length in mammals. Science 325, 866–870 (2009).
Gottlieb, D.J., O'Connor, G.T. & Wilk, J.B. Genome-wide association of sleep and circadian phenotypes. BMC Med. Genet. 8 (Suppl. 1), S9 (2007).
Gottlieb, D.J. et al. Novel loci associated with usual sleep duration: the CHARGE Consortium Genome-Wide Association Study. Mol. Psychiatry 20, 1232–1239 (2015).
Byrne, E.M. et al. A genome-wide association study of sleep habits and insomnia. Am. J. Med. Genet. B. Neuropsychiatr. Genet. 162B, 439–451 (2013).
Allebrandt, K.V. et al. A KATP channel gene effect on sleep duration: from genome-wide association studies to function in Drosophila. Mol. Psychiatry 18, 122–132 (2013).
Gehrman, P.R., Keenan, B.T., Byrne, E.M. & Pack, A.I. Genetics of sleep disorders. Psychiatr. Clin. North Am. 38, 667–681 (2015).
Sehgal, A. & Mignot, E. Genetics of sleep and sleep disorders. Cell 146, 194–207 (2011).
Lane, J.M. et al. Genome-wide association analysis identifies novel loci for chronotype in 100,420 individuals from the UK Biobank. Nat. Commun. 7, 10889 (2016).
Jones, S.E. et al. Genome-wide association analyses in 128,266 individuals identifies new morningness and sleep duration loci. PLoS Genet. 12, e1006125 (2016).
Hu, Y. et al. GWAS of 89,283 individuals identifies genetic variants associated with self-reporting of being a morning person. Nat. Commun. 7, 10448 (2016).
Pemberton, R. & Fuller Tyszkiewicz, M.D. Factors contributing to depressive mood states in everyday life: a systematic review. J. Affect. Disord. 200, 103–110 (2016).
Foral, P., Knezevich, J., Dewan, N. & Malesker, M. Medication-induced sleep disturbances. Consult Pharm. 26, 414–425 (2011).
Rosenberg, R.P. Clinical assessment of excessive daytime sleepiness in the diagnosis of sleep disorders. J. Clin. Psychiatry 76, e1602 (2015).
Gonnissen, H.K. et al. Sleep duration, sleep quality and body weight: parallel developments. Physiol. Behav. 121, 112–116 (2013).
Kurant, E. et al. Dorsotonals/homothorax, the Drosophila homologue of meis1, interacts with extradenticle in patterning of the embryonic PNS. Development 125, 1037–1048 (1998).
Casares, F. & Mann, R.S. Control of antennal versus leg development in Drosophila. Nature 392, 723–726 (1998).
Hisa, T. et al. Hematopoietic, angiogenic and eye defects in Meis1 mutant animals. EMBO J. 23, 450–459 (2004).
Davidson, S., Miller, K.A., Dowell, A., Gildea, A. & Mackenzie, A. A remote and highly conserved enhancer supports amygdala specific expression of the gene encoding the anxiogenic neuropeptide substance-P. Mol. Psychiatry 11 323, 410–421 (2006).
Oh-hashi, K., Naruse, Y., Amaya, F., Shimosato, G. & Tanaka, M. Cloning and characterization of a novel GRP78-binding protein in the rat brain. J. Biol. Chem. 278, 10531–10537 (2003).
Erhardt, A. et al. Replication and meta-analysis of TMEM132D gene variants in panic disorder. Transl. Psychiatry 2, e156 (2012).
Sklar, P. et al. Whole-genome association study of bipolar disorder. Mol. Psychiatry 13, 558–569 (2008).
Edwards, A.C. et al. Genome-wide association study of comorbid depressive syndrome and alcohol dependence. Psychiatr. Genet. 22, 31–41 (2012).
Han, K.E. et al. Pathogenesis and treatments of TGFBI corneal dystrophies. Prog. Retin. Eye Res. 50, 67–88 (2016).
Bradfield, J.P. et al. A genome-wide meta-analysis of six type 1 diabetes cohorts identifies multiple associated loci. PLoS Genet. 7, e1002293 (2011).
Patry, M. et al. βig-h3 represses T-cell activation in type 1 diabetes. Diabetes 64, 4212–4219 (2015).
Han, B. et al. TGFBI (βIG-H3) is a diabetes-risk gene based on mouse and human genetic studies. Hum. Mol. Genet. 23, 4597–4611 (2014).
Poelmans, G., Buitelaar, J.K., Pauls, D.L. & Franke, B. A theoretical molecular network for dyslexia: integrating available genetic findings. Mol. Psychiatry 16, 365–382 (2011).
Dalal, J. et al. Translational profiling of hypocretin neurons identifies candidate molecules for sleep regulation. Genes Dev. 27, 565–578 (2013).
Yelin-Bekerman, L. et al. Hypocretin neuron–specific transcriptome profiling identifies the sleep modulator Kcnh4a. eLife 4, e08638 (2015).
Mackiewicz, M. et al. Macromolecule biosynthesis: a key function of sleep. Physiol. Genomics 31, 441–457 (2007).
Takahama, K. et al. Pan-neuronal knockdown of the c-Jun N-terminal kinase (JNK) results in a reduction in sleep and longevity in Drosophila. Biochem. Biophys. Res. Commun. 417, 807–811 (2012).
Farh, K.K. et al. Genetic and epigenetic fine mapping of causal autoimmune disease variants. Nature 518, 337–343 (2015).
Ward, L.D. & Kellis, M. HaploReg v4: systematic mining of putative causal variants, cell types, regulators and target genes for human complex traits and disease. Nucleic Acids Res. 44 D1, D877–D881 (2016).
Hamdan, F.F. et al. De novo mutations in FOXP1 in cases with intellectual disability, autism, and language impairment. Am. J. Hum. Genet. 87, 671–678 (2010).
Fan, Y., Newman, T., Linardopoulou, E. & Trask, B.J. Gene content and function of the ancestral chromosome fusion site in human chromosome 2q13–2q14.1 and paralogous regions. Genome Res. 12, 1663–1672 (2002).
Fan, Y., Linardopoulou, E., Friedman, C., Williams, E. & Trask, B.J. Genomic structure and evolution of the ancestral chromosome fusion site in 2q13–2q14.1 and paralogous regions on other human chromosomes. Genome Res. 12, 1651–1662 (2002).
Wang, J., Duncan, D., Shi, Z. & Zhang, B. WEB-based GEne SeT AnaLysis Toolkit (WebGestalt): update 2013. Nucleic Acids Res. 41, W77–W83 (2013).
Cade, B.E. et al. Common variants in DRD2 are associated with sleep duration: the CARe consortium. Hum. Mol. Genet. 25, 167–179 (2016).
Loh, P.R. et al. Efficient Bayesian mixed-model analysis increases association power in large cohorts. Nat. Genet. 47, 284–290 (2015).
Bulik-Sullivan, B.K. et al. LD Score regression distinguishes confounding from polygenicity in genome-wide association studies. Nat. Genet. 47, 291–295 (2015).
Finucane, H.K. et al. Partitioning heritability by functional annotation using genome-wide association summary statistics. Nat. Genet. 47, 1228–1235 (2015).
Zhu, X. et al. Meta-analysis of correlated traits via summary statistics from GWASs with an application in hypertension. Am. J. Hum. Genet. 96, 21–36 (2015).
Mignot, E. Sleep, sleep disorders and hypocretin (orexin). Sleep Med. 5 (Suppl. 1), S2–S8 (2004).
Thompson, M.D., Xhaard, H., Sakurai, T., Rainero, I. & Kukkonen, J.P. OX1 and OX2 orexin/hypocretin receptor pharmacogenetics. Front. Neurosci. 8, 57 (2014).
Herring, W.J. et al. Suvorexant in patients with insomnia: results from two 3-month randomized controlled clinical trials. Biol. Psychiatry 79, 136–148 (2016).
Shieh, B.H. & Niemeyer, B. A novel protein encoded by the InaD gene regulates recovery of visual transduction in Drosophila. Neuron 14, 201–210 (1995).
Peirson, S.N. et al. Microarray analysis and functional genomics identify novel components of melanopsin signaling. Curr. Biol. 17, 1363–1372 (2007).
Bécamel, C. et al. The serotonin 5-HT2A and 5-HT2C receptors interact with specific sets of PDZ proteins. J. Biol. Chem. 279, 20257–20266 (2004).
Sharpley, A.L., Elliott, J.M., Attenburrow, M.J. & Cowen, P.J. Slow wave sleep in humans: role of 5-HT2A and 5-HT2C receptors. Neuropharmacology 33, 467–471 (1994).
Rosenberg, R. et al. APD125, a selective serotonin 5-HT2A receptor inverse agonist, significantly improves sleep maintenance in primary insomnia. Sleep 31, 1663–1671 (2008).
Winkelmann, J. et al. Genome-wide association study of restless legs syndrome identifies common variants in three genomic regions. Nat. Genet. 39, 1000–1006 (2007).
Xiong, L. et al. MEIS1 intronic risk haplotype associated with restless legs syndrome affects its mRNA and protein expression levels. Hum. Mol. Genet. 18, 1065–1074 (2009).
Schulte, E.C. et al. Targeted resequencing and systematic in vivo functional testing identifies rare variants in MEIS1 as significant contributors to restless legs syndrome. Am. J. Hum. Genet. 95, 85–95 (2014).
Spieler, D. et al. Restless legs syndrome–associated intronic common variant in Meis1 alters enhancer function in the developing telencephalon. Genome Res. 24, 592–603 (2014).
Moore, H. IV et al. Periodic leg movements during sleep are associated with polymorphisms in BTBD9, TOX3/BC034767, MEIS1, MAP2K5/SKOR1, and PTPRD. Sleep 37, 1535–1542 (2014).
Winkelmann, J. et al. Genome-wide association study identifies novel restless legs syndrome susceptibility loci on 2p14 and 16q12.1. PLoS Genet. 7, e1002171 (2011).
Allen, R.P., Barker, P.B., Horská, A. & Earley, C.J. Thalamic glutamate/glutamine in restless legs syndrome: increased and related to disturbed sleep. Neurology 80, 2028–2034 (2013).
Bulik-Sullivan, B. et al. An atlas of genetic correlations across human diseases and traits. Nat. Genet. 47, 1236–1241 (2015).
Byrne, E.M., Gehrman, P.R., Trzaskowski, M., Tiemeier, H. & Pack, A.I. Genetic correlation analysis suggests association between increased self-reported sleep duration in adults and schizophrenia and type 2 diabetes. Sleep 39, 1853–1857 (2016).
Wulff, K., Dijk, D.J., Middleton, B., Foster, R.G. & Joyce, E.M. Sleep and circadian rhythm disruption in schizophrenia. Br. J. Psychiatry 200, 308–316 (2012).
Poulin, J. et al. Sleep habits in middle-aged, non-hospitalized men and women with schizophrenia: a comparison with healthy controls. Psychiatry Res. 179, 274–278 (2010).
Chouinard, S., Poulin, J., Stip, E. & Godbout, R. Sleep in untreated patients with schizophrenia: a meta-analysis. Schizophr. Bull. 30, 957–967 (2004).
Hattersley, A.T. & Tooke, J.E. The fetal insulin hypothesis: an alternative explanation of the association of low birthweight with diabetes and vascular disease. Lancet 353, 1789–1792 (1999).
Horikoshi, M. et al. New loci associated with birth weight identify genetic links between intrauterine growth and adult height and metabolism. Nat. Genet. 45, 76–82 (2013).
Ananthakrishnan, A.N. et al. Sleep duration affects risk for ulcerative colitis: a prospective cohort study. Clin. Gastroenterol. Hepatol. 12, 1879–1886 (2014).
Tasali, E., Leproult, R., Ehrmann, D.A. & Van Cauter, E. Slow-wave sleep and the risk of type 2 diabetes in humans. Proc. Natl. Acad. Sci. USA 105, 1044–1049 (2008).
Nedeltcheva, A.V. & Scheer, F.A. Metabolic effects of sleep disruption, links to obesity and diabetes. Curr. Opin. Endocrinol. Diabetes Obes. 21, 293–298 (2014).
Vgontzas, A.N. et al. Obesity without sleep apnea is associated with daytime sleepiness. Arch. Intern. Med. 158, 1333–1337 (1998).
Bixler, E.O. et al. Excessive daytime sleepiness in a general population sample: the role of sleep apnea, age, obesity, diabetes, and depression. J. Clin. Endocrinol. Metab. 90, 4510–4515 (2005).
Swanson, J.M. The UK Biobank and selection bias. Lancet 380, 110 (2012).
Collins, R. What makes UK Biobank special? Lancet 379, 1173–1174 (2012).
Sudlow, C. et al. UK Biobank: an open access resource for identifying the causes of a wide range of complex diseases of middle and old age. PLoS Med. 12, e1001779 (2015).
Allen, N.E., Sudlow, C., Peakman, T. & Collins, R. UK Biobank. UK Biobank data: come and get it. Sci. Transl. Med. 6, 224ed4 (2014).
Howie, B.N., Donnelly, P. & Marchini, J. A flexible and accurate genotype imputation method for the next generation of genome-wide association studies. PLoS Genet. 5, e1000529 (2009).
O'Connell, J. et al. Haplotype estimation for biobank-scale data sets. Nat. Genet. 48, 817–820 (2016).
Loh, P.R., Palamara, P.F. & Price, A.L. Fast and accurate long-range phasing in a UK Biobank cohort. Nat. Genet. 48, 811–816 (2016).
Marchini, J., Howie, B., Myers, S., McVean, G. & Donnelly, P. A new multipoint method for genome-wide association studies by imputation of genotypes. Nat. Genet. 39, 906–913 (2007).
Wellcome Trust Case Control Consortium. Genome-wide association study of 14,000 cases of seven common diseases and 3,000 shared controls. Nature 447, 661–678 (2007).
Chang, C.C. et al. Second-generation PLINK: rising to the challenge of larger and richer datasets. Gigascience 4, 7 (2015).
International HapMap Consortium. Integrating common and rare genetic variation in diverse human populations. Nature 467, 52–58 (2010).
Willer, C.J., Li, Y. & Abecasis, G.R. METAL: fast and efficient meta-analysis of genomewide association scans. Bioinformatics 26, 2190–2191 (2010).
Schizophrenia Working Group of the Psychiatric Genomics Consortium. Biological insights from 108 schizophrenia-associated genetic loci. Nature 511, 421–427 (2014).
Locke, A.E. et al. Genetic studies of body mass index yield new insights for obesity biology. Nature 518, 197–206 (2015).
Dupuis, J. et al. New genetic loci implicated in fasting glucose homeostasis and their impact on type 2 diabetes risk. Nat. Genet. 42, 105–116 (2010).
Acknowledgements
This research has been conducted using the UK Biobank Resource under application number 6818. We would like to thank the participants and researchers from the UK Biobank who contributed or collected data. This work was supported by US NIH grants R01DK107859 (R.S.), R21HL121728 (R.S.), F32DK102323 (J.M.L.), R01HL113338 (J.M.L., S.R. and R.S.), R01DK102696 (R.S. and F.A.J.L.S.), R01DK105072 (R.S. and F.A.J.L.S.), T32HL007567 (J.L.) and HG003054 (X.Z.), the University of Manchester (Research Infrastructure Fund), the Wellcome Trust (salary support for D.W.R. and A.L.) and UK Medical Research Council MC_UU_12013/5 (D.A.L.). Data on glycemic traits have been contributed by MAGIC investigators and were downloaded from http://www.magicinvestigators.org/. Data on coronary artery disease and myocardial infarction have been contributed by CARDIoGRAMplusC4D investigators and were downloaded from http://www.cardiogramplusc4d.org/. We thank the International Genomics of Alzheimer's Project (IGAP) for providing summary results data for these analyses.
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J.M.L., M.K.R. and R.S. designed the study. J.M.L., J.L., I.V. and R.S. performed genetic analyses. J.M.L. and R.S. wrote the manuscript, and all co-authors helped interpret data and reviewed and edited the manuscript, before approving its submission. R.S. is the guarantor of this work and, as such, had full access to all the data in the study and takes responsibility for the integrity of the data and the accuracy of the data analysis.
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Lane, J., Liang, J., Vlasac, I. et al. Genome-wide association analyses of sleep disturbance traits identify new loci and highlight shared genetics with neuropsychiatric and metabolic traits. Nat Genet 49, 274–281 (2017). https://doi.org/10.1038/ng.3749
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DOI: https://doi.org/10.1038/ng.3749
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