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There are numerous potential benefits of developing objective biomarkers of insufficient sleep including providing screening and diagnostic tools, increasing our understanding of poor sleep health, and supporting sleep-based countermeasures. We analyzed plasma metabolites in humans during two separate occasions of experimental sleep restriction to identify stable biomarkers of insufficient sleep, and assessed the association of these biomarkers with cognitive performance. Methods 12 healthy adults (6M/6F), aged 24±5y (mean±SD), completed two 18-day protocols separated by 10 days. For each 18-day protocol, participants maintained habitual 9h sleep schedules for two weeks at home, and then completed 4-day laboratory visits with sleep opportunities of: 9h on night 1, 5h on nights 2 and 3, and recovery sleep on night 4. Energy balanced diets were provided 2 days before and throughout laboratory visits. Blood was collected every 2h during scheduled wakefulness on days 1–2 (baseline) and days 3–4 (insufficient sleep), and was analyzed by untargeted liquid chromatography/mass-spectrometry. Sustained attention was assessed every 2 h during scheduled wakefulness with the Psychomotor Vigilance Test (PVT). Results After filtering, we detected 6,822 metabolites. Elastic-net regression identified 21 metabolites consistently altered by insufficient sleep in both 18-day protocols. We developed a logistic regression-based biomarker fingerprint of insufficient sleep using 11 of these 21 metabolites with consistent responses to insufficient sleep. This biomarker fingerprint has 74.4% accuracy and 0.822 (0.782–0.858; 95%CI) area under the receiver operator curve. Median reaction time was slower (P&amp;lt;0.05) during insufficient sleep versus baseline. Mediation analyses show our biomarker fingerprint accounts for ~6% of the slower reaction time. Conclusion We identified a plasma metabolomics-based biomarker of insufficient sleep with “good” performance. However, this biomarker only accounts for ~6% of reduced PVT performance during insufficient sleep. Our findings suggest that specific biomarkers for insufficient sleep versus cognitive deficits associated with insufficient sleep may be required. Support This work was supported by NIH R01HL132150, NIH K01HL145099, NIH F32DK111161, NIH/NCATS Colorado CTSA Grant UL1TR002535, Sleep Research Society Foundation 011-JP-16, and the University of Colorado Boulder Undergraduate Research Opportunities Grant.

0056 Identification of a Stable Human Metabolomics-Based Biomarker of Insufficient Sleep and its Association with Cognitive Performance