Effect of sleep deprivation on the human metabolome

Sarah Davies

Joo Ang

Victoria Revell

Ben Holmes

Katrin Ackermann+4 authors

Supplementary Material

Supporting Information:

Click here to view.

^{Faculty of Health and Medical Sciences, University of Surrey, Guildford GU2 7XH, United Kingdom;}

^{Cancer Research UK Cancer Therapeutics Unit, Division of Cancer Therapeutics, The Institute of Cancer Research, London SM2 5NG, United Kingdom; and}

^{Department of Forensic Molecular Biology, Erasmus MC University Medical Center Rotterdam, 3000 CA Rotterdam, The Netherlands}

^{To whom correspondence should be addressed. Email:}ku.ca.yerrus@eneks.d.

Edited by Joseph S. Takahashi, Howard Hughes Medical Institute, University of Texas Southwestern Medical Center, Dallas, TX, and approved June 6, 2014 (received for review February 12, 2014)

Author contributions: V.L.R., K.A., M.K., and D.J.S. designed research; S.K.D., J.E.A., V.L.R., B.H., A.M., F.P.R., B.M., and F.I.R. performed research; S.K.D., J.E.A., B.H., N.C., F.I.R., and D.J.S. analyzed data; and S.K.D., J.E.A., V.L.R., K.A., A.E.T., F.I.R., and D.J.S. wrote the paper.

^{Present address: School of Chemistry, University of St. Andrews, St. Andrews KY16 9ST, United Kingdom.}

^{F.I.R. and D.J.S. contributed equally to this work.}

Edited by Joseph S. Takahashi, Howard Hughes Medical Institute, University of Texas Southwestern Medical Center, Dallas, TX, and approved June 6, 2014 (received for review February 12, 2014)

Freely available online through the PNAS open access option.

Significance

Sleep restriction and circadian clock disruption are associated with metabolic disorders including obesity and diabetes; this association can be studied by using the powerful tool of metabolomics. By using liquid chromatography/MS metabolomics, we have characterized plasma metabolites that were significantly affected by acute sleep deprivation (mainly lipids and acylcarnitines), all increasing during sleep deprivation. Observed increased levels of serotonin, tryptophan, and taurine may explain the antidepressive effect of sleep deprivation and deserve further study. Clear daily rhythms were observed in most metabolites, with 24 h wakefulness mainly reducing the amplitude of these rhythms. Our results further the understanding of sleep/wake regulation and the associated metabolic processes, and will be vital when using metabolic profiling to identify robust biomarkers for disease states and drug efficacy.

Keywords: circadian rhythms, total sleep deprivation, melatonin, depression, biomarker

Significance

Abstract

Sleep restriction and circadian clock disruption are associated with metabolic disorders such as obesity, insulin resistance, and diabetes. The metabolic pathways involved in human sleep, however, have yet to be investigated with the use of a metabolomics approach. Here we have used untargeted and targeted liquid chromatography (LC)/MS metabolomics to examine the effect of acute sleep deprivation on plasma metabolite rhythms. Twelve healthy young male subjects remained in controlled laboratory conditions with respect to environmental light, sleep, meals, and posture during a 24-h wake/sleep cycle, followed by 24 h of wakefulness. Two-hourly plasma samples collected over the 48 h period were analyzed by LC/MS. Principal component analysis revealed a clear time of day variation with a significant cosine fit during the wake/sleep cycle and during 24 h of wakefulness in untargeted and targeted analysis. Of 171 metabolites quantified, daily rhythms were observed in the majority (n = 109), with 78 of these maintaining their rhythmicity during 24 h of wakefulness, most with reduced amplitude (n = 66). During sleep deprivation, 27 metabolites (tryptophan, serotonin, taurine, 8 acylcarnitines, 13 glycerophospholipids, and 3 sphingolipids) exhibited significantly increased levels compared with during sleep. The increased levels of serotonin, tryptophan, and taurine may explain the antidepressive effect of acute sleep deprivation and deserve further study. This report, to our knowledge the first of metabolic profiling during sleep and sleep deprivation and characterization of 24 h rhythms under these conditions, offers a novel view of human sleep/wake regulation.

Abstract

Circadian clocks control the timing of most daily biological processes, including cyclic changes in metabolism and the sleep/wake cycle (1). There is a clear link between the circadian timing system and metabolism (2 –4), with disrupted circadian rhythms, sleep restriction, and sleep deprivation associated with metabolic disorders (obesity, insulin resistance, diabetes) and cardiovascular disease (5 –8). The underlying mechanisms linking metabolic disease, circadian clock misalignment, and sleep restriction are the subject of current research, elucidation of which will require a global “systems” approach (9). Transcriptomic studies have shown that rhythmic gene expression may be affected by sleep restriction, sleep deprivation, and mistimed sleep (10 –12), but, as yet, no studies have directly investigated the effect that sleep and sleep deprivation may have on the metabolic profile. Metabolic profiling, or “metabolomics,” is the profiling of small-molecule metabolites and offers the potential to characterize specific metabolic phenotypes associated with disrupted circadian timing and sleep loss. Metabolomics has an advantage over other “omics” techniques, in that it directly samples the metabolic changes in an organism and integrates information from changes at the gene, transcript, and protein levels, as well as posttranslational modification (13). Additionally, metabolomics can provide insight into the combination of genotype and environmental effects, leading to its use in predicting responses to drugs in a “pharmacometabolomic” approach (14, 15).

Daily rhythms have been identified in the metabolomes of mice (16 –18) and humans (19). Recent studies conducted in constant routine conditions, in which the impact of exogenous factors such as light, food, posture, and sleep are minimized, have demonstrated endogenous circadian variation in the human metabolome (20, 21), with one study reporting that ∼15% of the metabolites quantified in human plasma and saliva showed circadian variation, particularly amino acids in saliva and fatty acids in plasma (20). These human study protocols aimed to specifically identify circadian variation in the metabolome. However, if metabolic profiling is to be applied in “real-life” clinical settings, and in the identification of robust biomarkers, time-of-day variation in the human metabolome also needs to be characterized, and the effect of sleep, wakefulness, light/dark conditions, and meals on these daily rhythms assessed. Previously, we have characterized significant time of day variation in ∼19% of the metabolite features detected (19). These metabolites included corticosteroids, bilirubin, amino acids, acylcarnitines, and lysophospholipids. Not all of these metabolites, however, have been found to show circadian variation in the constant routine studies (20), suggesting that some metabolites may be affected by light/dark, sleep/wake, or food.

The two-process model of sleep regulation proposes that sleep is driven by a homeostatic component (i.e., process S) and an endogenous circadian oscillator (i.e., process C) (22). The link between the homeostatic and circadian regulation of sleep and metabolic pathways remains ill-defined. Sleep restriction or total sleep deprivation has been shown to reduce the number and the amplitude of genes exhibiting circadian rhythmicity (10, 11, 23). The expression of genes affected by sleep restriction included circadian clock and sleep homeostasis genes, as well those associated with oxidative stress and metabolism. How these changes in the transcriptome translate into changes in the metabolome, however, remains unknown.

Metabolomics can be used not only to study homeostatic regulation but also system perturbations. The metabolic pathways involved in human sleep and during sleep deprivation have not yet been systematically studied by using a metabolomics approach. Since 24 h sleep deprivation primarily affects the homeostatic regulator of sleep, acute total sleep deprivation permits assessment of the metabolic basis of this homeostatic process. The aim of the present study was thus to characterize plasma metabolite rhythms using untargeted and targeted liquid chromatography (LC)/MS metabolomics in healthy male participants during a 24 h wake/sleep cycle followed by 24 h of wakefulness.

Click here to view.

Acknowledgments

The authors thank Daniel Barrett, Cheryl Isherwood, and the Surrey Clinical Research Centre medical, clinical, and research teams for their help with the clinical study. This work was supported in part by the Netherlands Forensic Institute, Netherlands Genomics Initiative/Netherlands Organization for Scientific Research within the framework of the Forensic Genomics Consortium Netherlands, the 6th Framework project EUCLOCK (018741), and UK Biotechnology and Biological Sciences Research Council Grant BB/I019405/1. D.J.S. is a Royal Society Wolfson Research Merit Award holder.

Acknowledgments

Footnotes

Conflict of interest statement: D.J.S. has received research support from Philips Lighting. D.J.S. and B.M. are codirectors of Stockgrand. V.L.R. is a scientific advisor to Lumie and has received research support from Philips Lighting.

This article is a PNAS Direct Submission.

This article contains supporting information online at www.pnas.org/lookup/suppl/doi:10.1073/pnas.1402663111/-/DCSupplemental.

Footnotes

References

1. Mohawk JA, Green CB, Takahashi JSCentral and peripheral circadian clocks in mammals. Annu Rev Neurosci. 2012;35:445–462.[Google Scholar]
2. Green CB, Takahashi JS, Bass JThe meter of metabolism. Cell. 2008;134(5):728–742.[Google Scholar]
3. Bass J, Takahashi JSCircadian integration of metabolism and energetics. Science. 2010;330(6009):1349–1354.[Google Scholar]
4. Eckel-Mahan K, Sassone-Corsi PMetabolism and the circadian clock converge. Physiol Rev. 2013;93(1):107–135.[Google Scholar]
5. Bass J, Turek FWSleepless in America: A pathway to obesity and the metabolic syndrome? Arch Intern Med. 2005;165(1):15–16.[PubMed][Google Scholar]
6. Spiegel K, Knutson K, Leproult R, Tasali E, Van Cauter ESleep loss: A novel risk factor for insulin resistance and type 2 diabetes. J Appl Physiol (1985) 2005;99(5):2008–2019.[PubMed][Google Scholar]
7. Scheer FA, Hilton MF, Mantzoros CS, Shea SAAdverse metabolic and cardiovascular consequences of circadian misalignment. Proc Natl Acad Sci USA. 2009;106(11):4453–4458.[Google Scholar]
8. Cappuccio FP, Cooper D, D’Elia L, Strazzullo P, Miller MASleep duration predicts cardiovascular outcomes: A systematic review and meta-analysis of prospective studies. Eur Heart J. 2011;32(12):1484–1492.[PubMed][Google Scholar]
9. Rey G, Reddy ABConnecting cellular metabolism to circadian clocks. Trends Cell Biol. 2013;23(5):234–241.[PubMed][Google Scholar]
10. Maret S, et al Homer1a is a core brain molecular correlate of sleep loss. Proc Natl Acad Sci USA. 2007;104(50):20090–20095.[Google Scholar]
11. Ackermann K, et al Effect of sleep deprivation on rhythms of clock gene expression and melatonin in humans. Chronobiol Int. 2013;30(7):901–909.[PubMed][Google Scholar]
12. Archer SN, et al Mistimed sleep disrupts circadian regulation of the human transcriptome. Proc Natl Acad Sci USA. 2014;111(6):E682–E691.[Google Scholar]
13. Raamsdonk LM, et al A functional genomics strategy that uses metabolome data to reveal the phenotype of silent mutations. Nat Biotechnol. 2001;19(1):45–50.[PubMed][Google Scholar]
14. Clayton TA, et al Pharmaco-metabonomic phenotyping and personalized drug treatment. Nature. 2006;440(7087):1073–1077.[PubMed][Google Scholar]
15. Holmes E, Wilson ID, Nicholson JKMetabolic phenotyping in health and disease. Cell. 2008;134(5):714–717.[PubMed][Google Scholar]
16. Minami Y, et al Measurement of internal body time by blood metabolomics. Proc Natl Acad Sci USA. 2009;106(24):9890–9895.[Google Scholar]
17. Fustin JM, et al Rhythmic nucleotide synthesis in the liver: Temporal segregation of metabolites. Cell Reports. 2012;1(4):341–349.[PubMed][Google Scholar]
18. Eckel-Mahan KL, et al Coordination of the transcriptome and metabolome by the circadian clock. Proc Natl Acad Sci USA. 2012;109(14):5541–5546.[Google Scholar]
19. Ang JE, et al Identification of human plasma metabolites exhibiting time-of-day variation using an untargeted liquid chromatography-mass spectrometry metabolomic approach. Chronobiol Int. 2012;29(7):868–881.[Google Scholar]
20. Dallmann R, Viola AU, Tarokh L, Cajochen C, Brown SAThe human circadian metabolome. Proc Natl Acad Sci USA. 2012;109(7):2625–2629.[Google Scholar]
21. Kasukawa T, et al Human blood metabolite timetable indicates internal body time. Proc Natl Acad Sci USA. 2012;109(37):15036–15041.[Google Scholar]
22. Borbély AAA two process model of sleep regulation. Hum Neurobiol. 1982;1(3):195–204.[PubMed][Google Scholar]
23. Möller-Levet CS, et al Effects of insufficient sleep on circadian rhythmicity and expression amplitude of the human blood transcriptome. Proc Natl Acad Sci USA. 2013;110(12):E1132–E1141.[Google Scholar]
24. Hinard V, et al Key electrophysiological, molecular, and metabolic signatures of sleep and wakefulness revealed in primary cortical cultures. J Neurosci. 2012;32(36):12506–12517.[Google Scholar]
25. Monti JMSerotonin control of sleep-wake behavior. Sleep Med Rev. 2011;15(4):269–281.[PubMed][Google Scholar]
26. Peñalva RG, et al Effect of sleep and sleep deprivation on serotonergic neurotransmission in the hippocampus: A combined in vivo microdialysis/EEG study in rats. Eur J Neurosci. 2003;17(9):1896–1906.[PubMed][Google Scholar]
27. Alfaro-Rodríguez A, González-Piña R, González-Maciel A, Arch-Tirado ESerotonin and 5-hydroxy-indole-acetic acid contents in dorsal raphe and suprachiasmatic nuclei in normal, malnourished and rehabilitated rats under 24 h of sleep deprivation. Brain Res. 2006;1110(1):95–101.[PubMed][Google Scholar]
28. Wu JC, Bunney WEThe biological basis of an antidepressant response to sleep deprivation and relapse: review and hypothesis. Am J Psychiatry. 1990;147(1):14–21.[PubMed][Google Scholar]
29. Wirz-Justice A, Van den Hoofdakker RHSleep deprivation in depression: What do we know, where do we go? Biol Psychiatry. 1999;46(4):445–453.[PubMed][Google Scholar]
30. Ressler KJ, Nemeroff CBRole of serotonergic and noradrenergic systems in the pathophysiology of depression and anxiety disorders. Depress Anxiety. 2000;12(suppl 1):2–19.[PubMed][Google Scholar]
31. Vaswani M, Linda FK, Ramesh SRole of selective serotonin reuptake inhibitors in psychiatric disorders: a comprehensive review. Prog Neuropsychopharmacol Biol Psychiatry. 2003;27(1):85–102.[PubMed][Google Scholar]
32. Elmenhorst D, Kroll T, Matusch A, Bauer ASleep deprivation increases cerebral serotonin 2A receptor binding in humans. Sleep. 2012;35(12):1615–1623.[Google Scholar]
33. Coppen A, Noguera RL-tryptophan in depression. Lancet. 1970;1(7656):1111.[PubMed][Google Scholar]
34. Turner EH, Loftis JM, Blackwell ADSerotonin a la carte: Supplementation with the serotonin precursor 5-hydroxytryptophan. Pharmacol Ther. 2006;109(3):325–338.[PubMed][Google Scholar]
35. Mohammed HS, Aboul Ezz HS, Khadrawy YA, Noor NANeurochemical and electrophysiological changes induced by paradoxical sleep deprivation in rats. Behav Brain Res. 2011;225(1):39–46.[PubMed][Google Scholar]
36. Jia F, et al Taurine is a potent activator of extrasynaptic GABA(A) receptors in the thalamus. J Neurosci. 2008;28(1):106–115.[Google Scholar]
37. Coulon P, Budde T, Pape H-CThe sleep relay—the role of the thalamus in central and decentral sleep regulation. Pflugers Arch. 2012;463(1):53–71.[PubMed][Google Scholar]
38. Altamura C, Maes M, Dai J, Meltzer HYPlasma concentrations of excitatory amino acids, serine, glycine, taurine and histidine in major depression. Eur Neuropsychopharmacol. 1995;5(suppl):71–75.[PubMed][Google Scholar]
39. Perry TL, et al Hereditary mental depression and Parkinsonism with taurine deficiency. Arch Neurol. 1975;32(2):108–113.[PubMed][Google Scholar]
40. Quay WBCircadian rhythm in rat pineal serotonin and its modifications by estrous cycle and photoperiod. Gen Comp Endocrinol. 1963;14:473–479.[PubMed][Google Scholar]
41. Vellan EJ, Gjessing LR, Stalsberg HFree amino acids in the pineal and pituitary glands of human brain. J Neurochem. 1970;17(5):699–701.[PubMed][Google Scholar]
42. Wheler GH, Weller JL, Klein DCTaurine: stimulation of pineal N-acetyltransferase activity and melatonin production via a beta-adrenergic mechanism. Brain Res. 1979;166(1):65–74.[PubMed][Google Scholar]
43. Huang W, Ramsey KM, Marcheva B, Bass JCircadian rhythms, sleep, and metabolism. J Clin Invest. 2011;121(6):2133–2141.[Google Scholar]
44. Miyagawa T, et al Abnormally low serum acylcarnitine levels in narcolepsy patients. Sleep. 2011;34(3):349–53A.[Google Scholar]
45. Miyagawa T, et al Effects of oral L-carnitine administration in narcolepsy patients: A randomized, double-blind, cross-over and placebo-controlled trial. PLoS ONE. 2013;8(1):e53707.[Google Scholar]
46. Kuratsune H, et al Low levels of serum acylcarnitine in chronic fatigue syndrome and chronic hepatitis type C, but not seen in other diseases. Int J Mol Med. 1998;2(1):51–56.[PubMed][Google Scholar]
47. Fukuda K, et al International Chronic Fatigue Syndrome Study Group The chronic fatigue syndrome: A comprehensive approach to its definition and study. Ann Intern Med. 1994;121(12):953–959.[PubMed][Google Scholar]
48. Plioplys AV, Plioplys SAmantadine and l-carnitine treatment of chronic fatigue syndrome. Neuropsychobiology. 1997;35(1):16–23.[PubMed][Google Scholar]
49. Tafti M, et al Deficiency in short-chain fatty acid β-oxidation affects theta oscillations during sleep. Nat Genet. 2003;34(3):320–325.[PubMed][Google Scholar]
50. Hughes ME, et al Harmonics of circadian gene transcription in mammals. PLoS Genet. 2009;5(4):e1000442.[Google Scholar]
51. Ashley DV, Barclay DV, Chauffard FA, Moennoz D, Leathwood PDPlasma amino acid responses in humans to evening meals of differing nutritional composition. Am J Clin Nutr. 1982;36(1):143–153.[PubMed][Google Scholar]
52. Fukushima H, et al Nocturnal branched-chain amino acid administration improves protein metabolism in patients with liver cirrhosis: comparison with daytime administration. JPEN J Parenter Enteral Nutr. 2003;27(5):315–322.[PubMed][Google Scholar]