Sleep

BackgroundWe aimed to identify metabolites and create risk scores for insomnia symptoms in U.S. Hispanic/Latino adults. MethodsWe analyzed data from 6,107 participants in the Hispanic Community Health Study/Study of Latinos, split into discovery (n=3,932) and replication datasets (n=2,175). Serum metabolites and the Womens Health Initiative Insomnia Rating Scale (WHIIRS) were collected at baseline. We examined the relationships between 768 metabolites and insomnia symptoms and suspected insomnia (WHIIRS[≥]9) using the discovery dataset, followed by replication. Metabolite risk scores (MRSs) were generated with LASSO regression and evaluated for replication. We assessed the relationships of replicated metabolite measures and MRS with sleep, cognitive, and psychological traits (cross-phenotypes). FindingsNine metabolites were associated with insomnia symptoms in the discovery study, with two of these being replicated. Lower levels of hydrocinnamate and indolepropionate correlated with increased insomnia symptoms. We developed MRS for insomnia symptoms with replication. Various associations were observed between the two metabolites, 2 MRS, and cross-phenotypes. For instance, the WHIIRS MRS was associated with a higher risk of mild cognitive impairment (MCI) seven years later (OR:1.58, 95%CI:1.43-1.74 per 1 SD increase in MRS). InterpretationThe metabolomic profile associated with insomnia symptoms encompasses diet and gut microbiome metabolites. This study identified specific metabolites linked to insomnia that are also related to comorbidities, such as a higher risk of developing MCI during follow-up, suggesting a shared mechanism. FundingGrants from various National Institutes of Health and the JLH Foundation supported the work. Research in contextO_ST_ABSEvidence before this studyC_ST_ABSInsomnia affects 30-36% of individuals, with clinical insomnia estimated at 6-10%, and it is more severe among Hispanics, who also face higher risks for cognitive decline and cardiovascular disease. While previous metabolomics studies have investigated sleep disorders, most have focused on sleep apnea or sleep duration, not insomnia. The few studies that focus on insomnia were limited by small sample sizes or co-occurring psychiatric conditions. Only two large-scale studies linked insomnia symptoms to specific metabolites, but neither examined these associations in Hispanics or their connection to cognitive decline--gaps this study aims to address using data from the HCHS/SOL cohort. Added value of this studyWe identified nine metabolites related to insomnia symptoms, with two--hydrocinnamate and indolepropionate--being replicated. We also created and validated metabolite risk scores (MRS), which predicted a higher likelihood of developing mild cognitive impairment (MCI) seven years later. These results provide new insights into the metabolic pathways connecting insomnia and cognitive decline in a high-risk Hispanic population. Implications of all the available evidenceOur findings indicate that insomnia symptoms are linked to specific metabolic changes, some of which may also play a role in cognitive decline. Identifying metabolites related to diet and the gut microbiome points to biological pathways that could be modified through lifestyle or therapeutic interventions. The metabolite risk scores (MRS) developed in this study showed links with mild cognitive impairment (MCI) over time, suggesting their potential usefulness in understanding long-term health risks associated with sleep disturbances. These results encourage further research into the role of metabolomics in sleep and cognitive health, especially in high-risk populations like Hispanics.

Study ObjectivesIdiopathic hypersomnia (IH) is a central nervous system hypersomnia frequently accompanied by autonomic symptoms, yet objective physiological data are limited. We sought to characterize autonomic nervous system (ANS) dysfunction in IH using nocturnal heart rate variability (HRV) and diurnal autonomic reflex testing (ART), compared to individuals with type 1 narcolepsy (NT1) and healthy controls (HCs). MethodsTwenty-four adults with IH, 10 with NT1, and 14 HCs underwent overnight video polysomnography with HRV analyses in time and frequency domains during stable slow-wave sleep and REM sleep. Comprehensive ART included sympathetic adrenergic (head-up tilt (HUT), Valsalva BP responses), parasympathetic cardiovagal (HRV to deep breathing, Valsalva ratio), and sudomotor (Q-Sweat) measures. ResultsIH participants were predominantly female, with over half reporting long sleep duration. Compared to NT1 and HC, participants with IH demonstrated a greater magnitude of orthostatic tachycardia on tilt ({Delta}HR 41.0 {+/-} 16.3 vs. 26.3 {+/-} 9.3 vs. 30.8 {+/-} 9.3 bpm, p = 0.0086), as well as frequent sudomotor dysfunction (64.3%). IH participants demonstrated greater nocturnal and REM HR with reduced parasympathetic indices during REM, indicating diminished vagal modulation compared with HCs ConclusionsIH is characterized by a distinct pattern of autonomic dysfunction, including pronounced orthostatic tachycardia, frequent sudomotor abnormalities, and reduced parasympathetic activity during sleep. These findings provide objective physiological evidence of ANS involvement in IH and delineate features that distinguish IH from NT1 and HCs.

Type 1 narcolepsy (NT1), a disorder caused by the loss of hypocretin/orexin transmission, is characterized by daytime sleepiness and symptoms where Rapid Eye Movement (REM) sleep, a state normally occurring from middle to late in the night, can intermingle with wakefulness. This results in cataplexy and sleep paralysis, episodes of muscle paralysis when awake, or in the generation of dream-like hallucinations and vivid dreaming, periods of visual imagery or sensory experiences that occur while awake, notably when falling asleep (hypnagogic hallucinations) or lingering dreams with over-realistic recall. Using deep learning of nocturnal sleep polysomnography (PSG) signals (EEG, EMG and EOG) applied to sleep stage scoring, we found that NT1 shows abnormally short wake to REM sleep transitions and occurrences of abnormal sleep stages probabilities of wake, REM sleep and N1 (very light NREM) sleep abnormally co-occurs (sleep stage mixing). Interestingly, although presence of these during sleep enables NT1 diagnosis with performances similar to gold standard diagnostic procedure, the multiple sleep latency test (MSLT), the cortical localization of these dissociations remains unclear. In this work, we used electrode specific predictions of sleep stages to explore if these are global or observed at the local cortical level. Surprisingly, although sleep stage mixing was preeminent between REM sleep, N1 and wake across all electrodes, it was found to fluctuate across locations, with stronger fluctuations found in frontal and central locations, notably in the dominant (left) hemisphere. The strongest single discriminator for NT1 was N1-REM stage mixing across central electrodes (C3-C4), showing 4.3-fold higher dissociation in NT1 patients (Cohens d = 0.61). Analysis of sleep stage dissociations across varying time scales revealed that windows lasting several minutes were most predictive of NT1 status, aligning with the duration of clinically reported symptoms of dissociated REM sleep in narcolepsy. Local N1-W-REM sleep dissociations correlated with CSF orexin/hypocretin levels and severity as measured using MSLT. The predominance of stage mixing in frontal and central regions, areas typically associated with executive and motor control, may contribute to the partial preservation of awareness during dissociated REM phenomena. Further, self-reports of hypnagogic hallucinations correlated best with dissociations involving occipital locations, in agreement with its usual visual content. Coherence analysis was also conducted but did not reveal additional insight. These results suggest that orexin deficiency destabilizes REM sleep organization across cortical projection area contributing both to REM sleep dissociation and to abnormal state transitions observed in NT1.

Actigraphy is widely used for long-term sleep monitoring, but established sleep-wake scoring algorithms often require parameter tuning, which is commonly performed manually and can reduce reproducibility. In this study, a grid-search-based calibration framework is presented for established actigraphy algorithms and evaluate whether it can serve as a practical alternative to manual tuning. The method was evaluated using two datasets: a multi-subject polysomnography-validated actigraphy dataset and a self-collected dual-device dataset. In the polysomnography-validated dataset, grid-search optimization produced performance patterns similar to manual parameter selection, while slightly improving detection of sleep onset and sleep offset and yielding modest gains in wake-sensitive metrics. In the dual-device dataset, consensus and majority voting were useful for reducing the influence of brief wake episodes occurring within the main sleep period, including micro-awakenings that can fragment sleep predictions across individual algorithms. Overall, these findings show that grid-search can replace manual parameter tuning with a more explicit and reproducible procedure while providing small improvements in sleep timing estimation and benefiting ensemble-based handling of within-sleep wakefulness.

Background/ObjectivesElectromyography (EMG), video-polysomnography (vPSG), and wrist actigraphy are each used to develop diagnostic algorithms for Rapid eye movement sleep behavior disorder (RBD). However, the extent to which they capture overlapping versus distinct motor phenomena remains unknown. We evaluated the respective contributions of actigraphy, EMG and vPSG to the measurement of REM-sleep motor activity. MethodsSeventeen adults with RBD (Mount Sinai n = 9; Stanford n = 8) and eight control participants from an open Newcastle dataset underwent vPSG and concomitant wrist actigraphy. Flexor digitorum superficialis EMG activity and video-detected movements were manually scored in 3-second mini epochs. Actigraphy was quantified using an acceleration-magnitude-based activity count model. Statistical and agreement analyses were performed to assess the motor events captured by all three, any two, or by each modality independently during REM sleep. ResultsIn participants with RBD, actigraphy-derived movement load was significantly higher during REM sleep than during non-REM stages, a pattern not observed in control participants. Across 12,941 3-second mini epochs, EMG, actigraphy, and video detected 1,703, 1,613, and 811 motor events, of which 413 were detected concurrently by all three modalities. Pairwise agreement was moderate and increased from EMG-actigraphy ({kappa} = 0.27 {+/-} 0.10) to actigraphy-video ({kappa} = 0.41 {+/-} 0.12) and EMG-video ({kappa} = 0.45 {+/-} 0.15). Of EMG-detected events, 49.0% were also detected by actigraphy; of actigraphy-detected events, 37.2% were detected by EMG and 34.9% by video. Actigraphy activity counts were highest for events detected by all three modalities and lowest for actigraphy-only events. ConclusionActigraphy-measured REM-related motor activity was elevated in RBD but not in controls. EMG, actigraphy, and video captured partially overlapping motor events in RBD patient, with actigraphy showing the highest sensitivity and manually scored video the lowest.

Excessive daytime sleepiness (EDS) is a heterogeneous phenotype with little known of its genetic basis. Large-scale genome-wide association studies (GWAS) have reported genomic loci associated with EDS, though since most of these are non-coding, the causal gene(s) underlying the association are not known. Additionally, the cell types in which these genes exert their effects on sleep have not been functionally explored in vivo. Here, we employed a chromatin-based variant-to-gene mapping approach to first implicate candidate effector genes at EDS GWAS loci in human-derived neural and glial cell lines. Subsequent cell type-specific RNAi knockdown of orthologous genes using neural and glial GAL4 drivers in Drosophila confirmed cell-specific regulation of sleep by these GWAS-implicated effector genes. Among these, ruby (ortholog to AP3B2), a component of the AP-3 vesicular trafficking complex emerged as a robust sleep regulator. Targeted knockdown in flies localized ruby function to astrocyte-like glia, where loss of ruby increased sleep duration. The conserved role of ruby/ ap3b2 was validated in zebrafish where CRISPR-mediated loss increased daytime sleep. Together, these findings show that physical variant-to-gene mapping predicted cell-type-specific gene function for complex sleep traits and revealed ruby/AP3B2 as a conserved glial regulator of sleep and arousal. This work provides a generalizable framework for connecting non-coding GWAS variants and their corresponding effector genes to identify novel and highly conserved regulators of sleep.

ObjectiveIsolated rapid-eye-movement sleep behavior disorder is a prodromal marker of synucleinopathies. However, most cases remain undiagnosed due to the insufficient predictive value of questionnaires and limited access to confirmatory video-polysomnography. We assessed a two-stage screening strategy combining a brief questionnaire on rapid-eye-movement sleep behavior disorder symptoms and other prodromes with wrist actigraphy across multiple case-control cohorts. MethodsParticipants aged 40-80 without neurodegenerative disease were recruited from five cohorts; all cases were confirmed by video-polysomnography. The questionnaire was administered to 289 participants, and 236 underwent [≥]14 nights of home wrist actigraphy. The wearable-based algorithm was built on four movement features (mean motor activity, activity index, short or long immobile bouts, twitch activity). Models were trained with nested cross-validation using XGBoost. ResultsThe full retrospective cohort included 396 participants (99 cases, 297 controls; mean age 64 {+/-} 11; 55% male). The dream enactment question alone achieved an area under the curve of 0.85, which improved to 0.86 using the four-item questionnaire. Actigraphy alone achieved 82% sensitivity and 84% specificity. In the subgroup completing both assessments (75 cases, 54 controls), the two-stage protocol--questionnaire followed by actigraphy--yielded 68% sensitivity and 100% specificity using the dream-enactment question alone, and 73% sensitivity and 100% specificity using the four-item questionnaire. InterpretationA two-stage protocol combining questionnaire and actigraphy demonstrated high specificity and good sensitivity for detecting isolated rapid-eye-movement sleep behavior disorder in this multicenter cohort. This low-cost, scalable strategy is compatible with widely used wearable devices and warrants validation in community-based populations.

ObjectivesChronic insomnia (INS) is particularly prevalent in older adults and females. Sex-and age-related differences in neurophysiological markers of sleep quality (sleep spindles and slow-wave activity [SWA]) may underlie differential vulnerability to INS. This study investigated the effects of sex and insomnia on spindle and SWA beyond aging, to better understand the mechanistic differences contributing to the higher prevalence of INS in females. MethodsAfter a habituation night, one night of sleep assessed with polysomnography was analyzed in 222 adults (aged 18-82) including 119 INS (71% female) and 103 healthy sleepers (HS; 61% female). Spindle density, slow oscillation (SO) density, relative sigma power and SWA were derived during NREM sleep. Age, group, sex, and group-by-sex interactions were examined, with age as a covariate. ResultsAge, insomnia, and sex each contributed uniquely to NREM oscillatory activity. INS primarily reduced spindle and SO density, while sex accounted for differences in SWA. While SWA was higher in females overall, sex differences were not significant within the INS or HS groups. Female INS reported highest rates of insomnia severity as well as lower sigma power than males in the INS group. Spindle and SO density deficits were also present in female INS relative to female HS, as well as male INS relative to male HS. ConclusionsThe combination of reduced sigma power in females with insomnia relative to their male counterparts, as well as less spindle and SO density compared to female healthy sleepers may contribute to greater insomnia severity in females. Statement of SignificanceInsomnia is a growing public health concern that is more commonly reported in females, yet the neural mechanisms underlying this sex difference remain poorly understood. Our findings suggest that specific markers of sleep quality are disproportionately disrupted in females with insomnia, potentially contributing to greater vulnerability and symptom severity. These results provide new insight into how sex influences the neurophysiology of insomnia disorder and identify oscillatory markers that could serve as targets for personalized interventions. Future research should investigate whether these alterations represent persistent dysfunction or reversible changes, which could advance understanding of the biological basis of insomnia and inform strategies to improve sleep health in at-risk populations.

Background Narcolepsy is a rare, lifelong neurological disorder that often begins in childhood or adolescence. Diagnosis is frequently delayed because current diagnostic testing relies on specialist in-patient sleep investigations: overnight polysomnography (PSG) followed by a multiple sleep latency test (MSLT), interpreted according to International Classification of Sleep Disorders criteria (ICSD-3-TR). These investigations are expensive, labour intensive, and available in a limited number of centres, contributing to delays and inequity of access. Automated analysis of sleep-stage probabilities (hypnodensity) using neural networks has shown promising diagnostic performance in research cohorts but still requires hospital-based PSG acquisition. The Dreem 3 headband (DH) is a comfortable, dry-montage EEG device designed for home use. Combined with its proprietary machine learning classification of sleep stages, it may offer accurate ambulatory sleep physiology assessments and support clinical decision making. Methods This was a single-centre, prospective, observational study recruiting 60 participants aged 10 to 35 years undergoing investigation for hypersomnolence within GSTT sleep services and scheduled for PSG and MSLT as part of routine care. Exclusion criteria included physician-diagnosed medical or psychiatric disorder that could independently account for excessive daytime sleepiness; and/ or regular use of prescribed or recreational medication known to affect sleep architecture. Participants first wore the DH at home for five weeknights, followed by a continuous 48-hour weekend recording using two devices in rotation. They then underwent routine in-patient PSG and MSLT. PSG and MSLT were interpreted according to ICSD-3 by an experienced sleep physician and a final diagnosis determined by a sleep physiology consultant. The primary outcome is accuracy of ambulatory DH-based assessment of sleep physiology and subsequent diagnosis of sleep disorders. We evaluate proprietary and in-house developed machine learning methods to detect SOREM epochs and predict narcolepsy diagnosis from PSG, PSG+MSLT and DH data. All algorithmic outcomes will be compared to clinical outcomes derived from current clinical standard of care. Discussion This study will provide proof-of-concept evidence for a home-based wearable EEG approach to narcolepsy diagnosis. Patient and public involvement work with young people with confirmed narcolepsy indicates high acceptability of the DH protocol: in a survey of ten young people, eight reported they would be willing to wear a sleep headband nightly at home for five days (two were unsure), and seven reported they would be willing to wear it continuously for 48 hours over a weekend (two were unsure; one said no). These findings informed the decision to restrict continuous wear to the weekend, reflecting feedback that daytime wear during school or work hours would be unacceptable. If validated, this approach could reduce delays to diagnosis, improve equity of access, and support development of a subsequent multicentre study. Trial registration IRAS Project ID: 321547. Registered October 2022. Recruitment was completed on 30 January 2026.

IntroductionSleep spindles are electroencephalographic elements characteristic of non-rapid eye movement sleep generated by thalamo-cortical interactions. Spindles have been linked to some of the cognitive benefits afforded by sleep and high spindle activity is associated with increased arousal threshold. Here, we demonstrate that targeting the thalamus with Transcranial Electrical Stimulation with Temporal Interference (TES-TI) can enhance spindle activity. Methods46 participants (24 {+/-} 9.5 years; 58.7% F) underwent thalamic TES-TI stimulation during daytime naps. Three stimulation protocols with 15kHz carrier frequency were tested during stage 2 of non-rapid eye movement sleep (N2): fixed difference frequency of 10 Hz (TES15kHz-TI10Hz), difference frequency matched to individual spindle peak (TES15kHz-TIPeak), and no difference frequency (TES15kHz). Spectral power in the spindle (sigma) band and integrated spindle activity (ISA) were compared before and during the stimulation, and across stimulation protocols. ResultsTES15kHz-TI10Hz stimulation was associated with a significant increase in sigma band power ({Delta}[x]STIM-PRE = 0.46 log10{micro}V2, p = 0.0042) and ISA ({Delta}[x]STIM-PRE = 4.064 {micro}V/s, p = 0.030). Cluster-based analysis localized the increase in sigma power across the entire scalp (p = 0.008). Linear mixed effects models showed that both sigma band power and ISA during stimulation increased significantly more in TES15kHz-TI10Hz compared to the other experimental condition. ConclusionsThis study provides evidence supporting the successful use of TES-TI targeting the thalamus to enhance sleep spindle activity. Stimulation at a fixed difference frequency of 10 Hz increased sigma band power and ISA, whereas neither stimulation matched to individual sigma band peak nor TES alone produced comparable effects. These promising results warrant further investigations into the cognitive and clinical impact of TES-TI, a non-invasive neuromodulation tool that can reach deep brain regions. Statement of significanceThis study provides evidence that thalamo-cortical networks, which are central to many physiological and pathological brain activities, can be modulated non-invasively in humans. More specifically, the findings show that transcranial electrical stimulation with temporal interference targeting the thalamus can selectively enhance sleep spindle activity. This work introduces a new strategy for precisely targeting sleep-generating mechanisms regulated by deep brain circuits without surgery or medication. Key next steps include determining whether this increase in spindle activity can positively impact cognition and assessing the translational potential of this approach for clinical populations.

Both short and long sleep duration have been associated with poor glycemic control and an increased risk of developing type 2 diabetes mellitus. Although sleep duration may differentially modify the effects of genetic risk factors for type 2 diabetes, this has not been systematically investigated. In the present study, we conducted genome-wide gene by sleep duration meta-analyses, separately assessing interactions of short and long sleep, for fasting glucose, fasting insulin, and hemoglobin A1c in up to 489,309 individuals without diabetes from seven different population groups. In total, 16 loci were identified to interact with sleep duration -- six with short sleep and ten with long sleep. Of these, four loci were identified through cross-population meta-analysis. Mapped genes exhibit pathway connections to pericyte apoptosis, NMDA receptor activity, the GLUT1 receptor, neurological health, and sleep architecture. Eleven loci (VRK2, PCDH7, TFAP2A, CAP2, PAPPA, ZCCHC2, MYH9, SGIP1, JAKMIP3, RRAS2, MAPT) have not been reported in previous glycemic trait genome-wide association studies. Interaction loci identify divergent biological mechanisms for short and long sleep duration influencing glycemic control, suggesting specific pathways of intervention for precision medicine approaches to diabetes prevention and management. Article HighlightsO_LIThe biological mechanisms of how sleep duration impacts type 2 diabetes pathogenesis and glycemic control are unclear. C_LIO_LIThis study reveals 16 loci (11 novel) that interact with either short or long sleep duration to influence hemoglobin A1c, fasting glucose, or fasting insulin. Short and long sleep duration loci were non-overlapping. C_LIO_LIRegulation of copper and diacylglycerol levels appear as distinct cellular mechanisms implicated by long and short sleep duration loci respectively. C_LIO_LIIdentified gene targets present insight for potential type 2 diabetes therapeutic design approaches related to JIP1-JNK interaction disruption, pericyte health, NMDA receptor activity, anti-inflammatory and leptin-enhancing dietary supplements, and serpins. C_LI

The transition from wake to stable sleep is characterized by multiple neural, physiological, and behavioral changes. How these changes may differ in individuals with difficulties falling asleep such as children with neurodevelopmental conditions is poorly understood. Here, we studied sleep initiation in >2000 nights recorded from 186 children who participated in the Simons Sleep Project (SSP). Data included simultaneous, synchronized recordings of actigraphy, electroencephalography (EEG), photoplethysmography (PPG), and skin temperature. We extracted multiple neural, physiological, and behavioral measures that are known to increase/decrease during the sleep initiation period including EEG delta (1-4Hz) power, movement counts, heart rate (HR), and skin temperature. Transitions from 20 minutes before sleep onset to 40 minutes after sleep onset were modeled with a sigmoid function enabling the quantification of transition timing, speed, and magnitude per measure. Individuals with longer sleep onset latencies (SOL) exhibited smaller increases in EEG delta power and skin temperature as well as smaller decreases in HR and activity counts. These findings indicate that difficulties falling asleep are associated with multiple forms of cortical, physiological, and behavioral hyperarousal that can be measured at home with wearable devices. Importantly, transition magnitudes were key to explaining differences in SOL across participants (26% explained variance) in contrast to transition speed or timing within the sleep initiation period (<13% explained variance). Longer SOL and weaker transitions were particularly prominent in children diagnosed with autism and/or attention deficit hyperactivity disorder (ADHD).

Objective: The narcolepsy type 1 (NT1) phenotype severity is heterogeneous, and the disease course is largely unknown. The 2009-10 H1N1-(Pandemrix(R))-vaccinations were followed by increased numbers of possibly more severe post-H1N1 NT1 cases but long-term prospective data on large, vaccinated cohorts are missing. Methods: 130 consecutive post-H1N1 NT1 cases (113/130 Pandemrix(R) -vaccinated) were prospectively followed up after approximately 5.5 years. Epworth Sleepiness Scale (ESS), cataplexy, hypnagogic hallucinations, sleep paralysis, PSG, MSLT, and BMI were evaluated. Phenotype severity predictors (hypocretin-1 deficiency severity <40 vs. 40-150 pg/ml; Pandemrix(R)- vaccination; disease duration) were tested in age and sex-adjusted multivariable regressions. Results: From baseline to follow-up, phenotype severity overall improved (milder symptoms, higher mean MSLT sleep latency (SL) and fewer SOREMPs, all p<0.001). Follow-up phenotype severity was strongly predicted by the same baseline measures. Females had worse ESS and cataplexy, men had higher BMI, and young individuals had lower mean MSLT SL and more SOREMPs. Severe hypocretin deficiency (<40 pg/ml) predicted baseline PSG SOREMPs and lower MSLT SL. Vaccinated individuals had more severe baseline PSG/MSLT measures but greater long-term symptom improvement, and vaccination no longer predicted PSG/MSLT severity at follow-up. Conclusion: The best prognostic factor for long-term NT1 phenotype severity is the earlier phenotype severity. Hypocretin deficiency severity also predicts parts of short-term but not long-term phenotype severity. Pandemrix(R)-vaccination is associated with initially more severe phenotype but larger long-term improvement i.e. a different clinical course than in unvaccinated NT1, although medication effects cannot be excluded. Our findings underscore heterogeneity in NT1 phenotype and disease trajectories.

Chronic short sleep (CSS) is an emerging public health issue that frequently begins in adolescence and is common among healthcare professionals and others engaged in shift work. Epidemiological studies associate CSS and sleep disruption with metabolic disorders, cardiovascular disease, cognitive decline, and heightened Alzheimers disease risk. Building on our prior findings that sleep deprivation perturbs proteostasis and activates endoplasmic reticulum (ER) stress pathways, we investigated the long-term consequences of CSS in young adult wild-type mice over the course of one year. Mice exposed to CSS displayed impaired cognition in hippocampal dependent tasks by 28 weeks of age, indicating emerging memory deficits. At the molecular level, CSS disrupted hippocampal proteostasis--particularly protein folding processes--and triggered ER stress and activation of the unfolded protein response (UPR). Importantly, disrupted proteostasis preceded the behavioral decline, with diminution of the key chaperone and UPR regulator BiP occurring at 20-22 weeks of age. CSS also increased markers of cellular stress and neuroinflammation while reducing the expression of proteins associated with memory function. Age also seemed to be a cellular stressor, causing a longitudinal increase in UPR, ISR, and neuroinflammation markers. Together, these results indicate that both chronic short sleep and age compromise proteostasis and promote neuroinflammation, contributing to progressive cognitive dysfunction.

Study ObjectivesNarcolepsy type 1 (NT1) is characterized by excessive daytime sleepiness and cataplexy. Previous studies have implicated the amygdala, thalamus, brainstem and hippocampus in the pathophysiology of NT1. We here aimed to examine more detailed subregional case-control differences in MRI-based segmentations of these brain regions to gain deeper insights. MethodsWe obtained 3T MRI brain scans from 54 NT1 patients (39 females, mean age 21.8 {+/-} 11.0 years, 51 with confirmed hypocretin-deficiency and three patients that had not performed this measure) and 114 healthy controls (77 females, mean age 23.2 {+/-} 9.0 years). Automated segmentation of the hippocampus, amygdala, thalamus, and brainstem was performed on T1-weighted MRI data using FreeSurfer. Case-control volume differences were tested using general linear models and permutation testing. The false discovery rate was controlled at 5% with the Benjamini-Hochberg procedure. ResultsThe analysis revealed no significant case-control differences for any of the subregions in the hippocampus, thalamus, amygdala and brainstem after correction for multiple testing. ConclusionsBased on a detailed automated MRI-based segmentation analyses in a relatively large national sample, NT1 patients had no significant changes in any amygdala, thalamus, brainstem or hippocampus subregions compared to controls. In the future large multi-site studies could be performed to achieve sufficient power to detect more subtle group differences.

Cannabis use is an increasingly common therapeutic for a variety of chronic diseases. In addition, people with sleep problems may self-medicate using cannabis products. However, genetic architecture of cannabis use and its shared genetic predispositions with sleep traits has not been systematically examined. We performed a meta-analysis of cannabis use within the All of Us and UK Biobank cohorts, consisting of 152,807 cases and 220,272 controls. Our meta-analysis identified 39 independent loci, including the previously reported CADM2 locus associated with cannabis use and replicating previous work. Additionally our associations include neuronal and sleep-regulating genes such as HTR1A, RAI1, SLC39A8, and NCAM1. Moreover, tissue-specific analyses revealed that the genetic architecture of cannabis use is heavily enriched within the central nervous system and specific brain cell types. In addition, we observed significant positive genetic correlations with clinical insomnia, insomnia-related medication usage, and objectively measured nighttime physical activity, alongside negative correlations with morningness chronotype and daytime activity. Fine-mapping and colocalization analyses identified shared genetic signals between cannabis use and clinical insomnia including a near-perfect colocalization at SLC39A8 and CADM2. Together, these results highlight the shared genetic risk between cannabis use and sleep disorders. Additionally, our findings indicate the importance of investigating the genetic effects of cannabis use as its use becomes more widespread, both recreationally and medicinally.

Rapid Eye Movement (REM) sleep makes up approximately 20% of sleep in the adult human and is altered in psychiatric and neurodegenerative conditions. REM sleep comprises two substates, during which eye movements do (phasic REM) and do not (tonic REM) occur. Tonic REM makes up 70-90% of REM sleep but its role in regulating brain function, mood and cognition remain underexplored. We investigated how seven nights of insufficient sleep (6 h time in bed), compared to sufficient sleep, alter periodic and aperiodic components of the phasic and tonic REM sleep electroencephalography (EEG), in 542 sleep recordings of 36 young adults. Associations between phasic and tonic REM sleep EEG and mood, cognitive performance, and overnight changes in cortical excitability as indexed by 1/f spectral slopes were assessed. Insufficient sleep predominantly affected tonic REM EEG components, specifically the density of theta, the amplitude, density, and frequency of alpha oscillations and the 1/f slope in the 30 to 45 Hz range. These changes associated with mood and cognitive performances, and with overnight reductions in cortical excitability. These results provide evidence for a role of tonic REM sleep in regulating mood and counteracting cognitive deterioration and excitability changes associated with insufficient sleep.

Mounting evidence shows sex-based differences in sleep experiences and outcomes, including the prevalence of insomnia disorder. However, the impact of biological sex on brain oscillations during sleep remains poorly understood, especially in the context of insomnia disorder. This is a notable gap, given that neurophysiological aspects of sleep are associated with brain health and overall sleep quality. We systematically reviewed and meta-analysed data from studies reporting spindle and slow wave activity in adults with and without insomnia disorder. We conducted systematic searches in PubMed, Embase, Scopus, and PsycInfo. Risk of bias was evaluated using the Cochrane Risk of Bias tool. Forty-three studies met our inclusion criteria, with thirteen studies of normal sleepers (N= 668) reporting sufficient data for random-effects meta-analyses. Compared with males, female normal sleepers had higher spindle density, sigma and delta power. Most studies recruited individuals with primary insomnia, and data pooling for insomnia and mixed groups was not possible due to insufficient statistical reporting. Moreover, group-by-sex interactions were limited, inconsistent, and varied across studies and sample characteristics. Further research is needed to explore sex-specific differences in sleep microarchitecture and their role in normal sleep and the manifestation of insomnia disorder.

Neuroligin-3 (NLGN3) was first identified as a risk gene associated with autism spectrum disorder (ASD). The initial variant, p.R451C, associating NLGN3 with ASD has been heavily investigated, yet little is known about the functional consequences of other NLGN3 variants. Furthermore, while most of the identified variants are present in males with maternally inherited variants from unaffected mothers, several de novo variants were observed in females, suggesting a possible functional difference between de novo and maternally inherited variants. To address the functional consequences of NLGN3 variants in vivo, we generated transgenic Drosophila models corresponding to one de novo variant (p.R175W) identified in one female proband, and two maternally inherited variants (p.R451C and p.R597W) identified in male probands. In Drosophila, loss of the fly homolog, Nlg3, altered sleep patterns, synaptic architecture, and vesicle dynamics, which were rescued by the expression of the human NLGN3Ref allele. When comparing the variants, the de novo p.R175W variant and the maternally inherited p.R451C variant altered synapse morphology and sleep patterns, with minimal effects on vesicle dynamics, and the p.R597W variant altered sleep and vesicle dynamics with minimal impact on synapse morphology. Using overexpression models, human NLGN3Ref altered sleep patterns and synaptic morphology. Moreover, the p.R175W variant exacerbated sleep phenotypes, and the p.R175W and p.R451C variants exacerbated synapse morphology phenotypes. Together, our findings suggest that de novo NLGN3 variants identified in females are likely gain-of-function, while maternally inherited variants have mixed loss-and gain-of-function effects. Moreover, the location of the variants may contribute to the distinct functional differences we observed. Some NLGN3 variants disrupt synaptic development, while other variants alter synaptic function, suggesting that NLGN3 variants have differential effects. These functional differences may provide insight into the heterogeneity of individuals with ASD. Author SummaryAutism spectrum disorder (ASD) is a common neurodevelopmental disorder. Mutations in the Neuroligin-3 (NLGN3) gene are associated with ASD but very few of these mutations have been characterized in animal models. Most of these mutations affect male individuals who maternally inherited their genetic mutation; however, more rarely female individuals may present with a genetic mutation that was not identified in either of the parents. Here, we utilized the fruit fly model to investigate how three different mutations, one mutation identified in a female and two mutations identified in males, affect the flys behavior and synapse development. We identified altered sleep patterns in some of our mutants which is consistent with sleep disturbances being highly comorbid with ASD. Additionally, we identified alterations in synapse development and function which is consistent with the role of NLGN3 in synapse formation and maturation. Together, our findings support that NLGN3 is important for regulating the synapse and mutations in this gene can alter its function. However, different mutations can have differential effects. This demonstrates the need to assess multiple variants simultaneously because each variant may have distinct functional significances.

Sleep architecture is essential for metabolic and cardiovascular health, yet the impact of day-to-day dietary variation on objective sleep physiology remains unclear. Using 4.8 thousand person-nights with real-time dietary logs and multi-stage wearable sleep recordings, we examined how prior-day nutrition relates to next-night sleep under free-living conditions. Higher fiber density was associated with increased restorative sleep, including +0.59 pp deep sleep, +0.76 pp REM sleep, -1.35 pp light sleep, and -1.14 bpm lower mean nocturnal heart rate. Greater plant diversity and higher whole-plant food intake were similarly associated with lower nocturnal heart rate (-0.72 to -0.94 bpm). Meal-timing behaviors primarily influenced sleep duration, sleep-onset latency, and autonomic tone: heavier evening meals were associated with +7.7 min longer total sleep time and +0.73 bpm higher nocturnal heart rate. In contrast, short-term variation in macronutrient energy distribution and micronutrient consumption showed no robust associations with sleep outcomes. When analyses were restricted to more extreme dietary contrasts, effect magnitudes increased while remaining directionally consistent. These findings indicate that routine daily dietary choices, particularly plant-forward composition and meal timing, have immediate and measurable effects on objective sleep architecture.