Background: There is no consistent evidence of specific gene(s) or molecular pathways that contribute to the pathogenesis, therapeutic intervention or diagnosis of chronic fatigue syndrome (CFS). While multiple studies support a role for genetic variation in CFS, genome-wide efforts to identify associated loci remain unexplored. We employed a novel convergent functional genomics approach that incorporates the findings from single-nucleotide polymorphism (SNP) and mRNA expression studies to identify associations between CFS and novel candidate genes for further investigation. Methods: We evaluated 116,204 SNPs in 40 CFS and 40 nonfatigued control subjects along with mRNA expression of 20,160 genes in a subset of these subjects (35 CFS subjects and 27 controls) derived from a population-based study. Results: Sixty-five SNPs were nominally associated with CFS (p < 0.001), and 165 genes were differentially expressed (≧4-fold; p ≤ 0.05) in peripheral blood mononuclear cells of CFS subjects. Two genes, glutamate receptor, ionotropic, kinase 2 (GRIK2) and neuronal PAS domain protein 2 (NPAS2), were identified by both SNP and gene expression analyses. Subjects with the G allele of rs2247215 (GRIK2) were more likely to have CFS (p = 0.0005), and CFS subjects showed decreased GRIK2 expression (10-fold; p = 0.015). Subjects with the T allele of rs356653 (NPAS2) were more likely to have CFS (p = 0.0007), and NPAS2 expression was increased (10-fold; p = 0.027) in those with CFS. Conclusion: Using an integrated genomic strategy, this study suggests a possible role for genes involved in glutamatergic neurotransmission and circadian rhythm in CFS and supports further study of novel candidate genes in independent populations of CFS subjects.
© 2011 S. Karger AG, Basel
- Chronic fatigue syndrome
- Genome-wide association
- Gene expression
- Glutamatergic neurotransmission
- Circadian rhythm
- Orexin signaling
- Klimas NG, Koneru AO: Chronic fatigue syndrome: inflammation, immune function, and neuroendocrine interactions. Curr Rheumatol Rep 2007;9:482–487.
- Togo F, Natelson BH, Cherniack NS, FitzGibbons J, Garcon C, Rapoport DM: Sleep structure and sleepiness in chronic fatigue syndrome with or without coexisting fibromyalgia. Arthritis Res Ther 2008;10:R56.
- Van Den Eede F, Moorkens G, Van Houdenhove B, Cosyns P, Claes SJ: Hypothalamic-pituitary-adrenal axis function in chronic fatigue syndrome. Neuropsychobiology 2007;55:112–120.
- Smith AK, Dimulescu I, Falkenberg VR, Narasimhan S, Heim C, Vernon SD, Rajeevan MS: Genetic evaluation of the serotonergic system in chronic fatigue syndrome. Psychoneuroendocrinology 2008;33:188–197.
- Sullivan PF, Evengard B, Jacks A, Pedersen NL: Twin analyses of chronic fatigue in a Swedish national sample. Psychol Med 2005;35:1327–1336.
- van de Putte EM, van Doornen LJ, Engelbert RH, Kuis W, Kimpen JL, Uiterwaal CS: Mirrored symptoms in mother and child with chronic fatigue syndrome. Pediatrics 2006;117:2074–2079.
- Kerr JR: Gene profiling of patients with chronic fatigue syndrome/myalgic encephalomyelitis. Curr Rheumatol Rep 2008;10:482–491.
- Rajeevan MS, Smith AK, Dimulescu I, Unger ER, Vernon SD, Heim C, Reeves WC: Glucocorticoid receptor polymorphisms and haplotypes associated with chronic fatigue syndrome. Genes Brain Behav 2007;6:167–176.
- Saiki T, Kawai T, Morita K, Ohta M, Saito T, Rokutan K, Ban N: Identification of marker genes for differential diagnosis of chronic fatigue syndrome. Mol Med 2008;14:599–607.
- Sorensen B, Jones JF, Vernon SD, Rajeevan MS: Transcriptional control of complement activation in an exercise model of chronic fatigue syndrome. Mol Med 2008;15:34–42.
- Harrison PJ, Weinberger DR: Schizophrenia genes, gene expression, and neuropathology: on the matter of their convergence. Mol Psychiatry 2005;10:40–68.
- Middleton FA, Rosenow C, Vailaya A, Kuchinsky A, Pato MT, Pato CN: Integrating genetic, functional genomic, and bioinformatics data in a systems biology approach to complex diseases: application to schizophrenia. Methods Mol Biol 2007;401:337–364.
- Vawter MP, Atz ME, Rollins BL, Cooper-Casey KM, Shao L, Byerley WF: Genome scans and gene expression microarrays converge to identify gene regulatory loci relevant in schizophrenia. Hum Genet 2006;119:558–570.
- Jiao H, Kaaman M, Dungner E, Kere J, Arner P, Dahlman I: Association analysis of positional obesity candidate genes based on integrated data from transcriptomics and linkage analysis. Int J Obes (Lond) 2008;32:816–825.
- Glinsky GV: Integration of HapMap-based SNP pattern analysis and gene expression profiling reveals common SNP profiles for cancer therapy outcome predictor genes. Cell Cycle 2006;5:2613–2625.
- Huang RS, Duan S, Bleibel WK, Kistner EO, Zhang W, Clark TA, Chen TX, Schweitzer AC, Blume JE, Cox NJ, Dolan ME: A genome-wide approach to identify genetic variants that contribute to etoposide-induced cytotoxicity. Proc Natl Acad Sci USA 2007;104:9758–9763.
- Kurian SM, Le-Niculescu H, Patel SD, Bertram D, Davis J, Dike C Yehyawi N, Lysaker P, Dustin J, Caligiuri M, Lohr J, Lahiri DK, Nurnberger JI Jr, Faraone SV, Geyer MA, Tsuang MT, Schork NJ, Salomon DR, Niculescu AB: Identification of blood biomarkers for psychosis using convergent functional genomics. Mol Psychiatry 2011;16:37–58.
- Le-Niculescu H, Patel SD, Bhat M, Kuczenski R, Faraone SV, Tsuang MT, McMahon FJ, Schork NJ, Nurnberger JI Jr, Niculescu AB 3rd: Convergent functional genomics of genome-wide association data for bipolar disorder: comprehensive identification of candidate genes, pathways and mechanisms. Am J Med Genet B Neuropsychiatr Genet 2009;150B:155–181.
- Le-Niculescu H, Kurian SM, Yehyawi N, Dike C, Patel SD, Edenberg HJ, Tsuang MT, Salomon DR, Nurnberger JI Jr, Niculescu AB: Identifying blood biomarkers for mood disorders using convergent functional genomics. Mol Psychiatry 2009;14:156–174.
- Niculescu AB, Le-Niculescu H: The P-value illusion: how to improve (psychiatric) genetic studies. Am J Med Genet B Neuropsychiatr Genet 2010;153B:847–849.
- Niculescu AB, Le-Niculescu H: Convergent functional genomics: what we have learned and can learn about genes, pathways, and mechanisms. Neuropsychopharmacology 2010;35:355–356.
- Achiron A, Gurevich M: Peripheral blood gene expression signature mirrors central nervous system disease: the model of multiple sclerosis. Autoimmun Rev 2006;5:517–522.
- Reeves WC, Wagner D, Nisenbaum R, Jones JF, Gurbaxani B, Solomon L, Papanicolaou DA, Unger ER, Vernon SD, Heim C: Chronic fatigue syndrome – a clinically empirical approach to its definition and study. BMC Med 2005;3:19.
- Fukuda K, Straus SE, Hickie I, Sharpe MC, Dobbins JG, Komaroff A: The chronic fatigue syndrome: a comprehensive approach to its definition and study. International Chronic Fatigue Syndrome Study Group. Ann Intern Med 1994;121:953–959.
- Reeves WC, Lloyd A, Vernon SD, Klimas N, Jason LA, Bleijenberg G, Evengard B, White PD, Nisenbaum R, Unger ER; International Chronic Fatigue Syndrome Study Group: Identification of ambiguities in the 1994 chronic fatigue syndrome research case definition and recommendations for resolution. BMC Health Serv Res 2003;3:25.
- Devlin B, Roeder K: Genomic control for association studies. Biometrics 1999;55:997–1004.
- Ojaniemi H, Evengard B, Lee DR, Unger ER, Vernon SD: Impact of RNA extraction from limited samples on microarray results. Biotechniques 2003;35:968–973.
- Chen R, Morgan AA, Dudley J, Deshpande T, Li L, Kodama K, Chiang AP, Butte AJ: FitSNPs: highly differentially expressed genes are more likely to have variants associated with disease. Genome Biol 2008;9:R170.
- Majer M, Welberg LA, Capuron L, Miller AH, Pagnoni G, Reeves WC: Neuropsychological performance in persons with chronic fatigue syndrome: results from a population-based study. Psychosom Med 2008;70:829–836.
- Reeves WC, Heim C, Maloney EM, Youngblood LS, Unger ER, Decker MJ, Jones JF, Rye DB: Sleep characteristics of persons with chronic fatigue syndrome and non-fatigued controls: results from a population-based study. BMC Neurol 2006;6:41.
- Van Houdenhove B, Kempke S, Luyten P: Psychiatric aspects of chronic fatigue syndrome and fibromyalgia. Curr Psychiatry Rep 2010;12:208–214.
- Kim SA, Kim JH, Park M, Cho IH, Yoo HJ: Family-based association study between GRIK2 polymorphisms and autism spectrum disorders in the Korean trios. Neurosci Res 2007;58:332–335.
- Zeng W, Gillis T, Hakky M, Djousse L, Myers RH, MacDonald ME, Gusella JF: Genetic analysis of the GRIK2 modifier effect in Huntington’s disease. BMC Neurosci 2006;7:62.
- Dorval KM, Wigg KG, Crosbie J, Tannock R, Kennedy JL, Ickowicz A, Pathare T, Malone M, Schachar R, Barr CL: Association of the glutamate receptor subunit gene GRIN2B with attention-deficit/hyperactivity disorder. Genes Brain Behav 2007;6:444–452.
- Arnold PD, Rosenberg DR, Mundo E, Tharmalingam S, Kennedy JL, Richter MA: Association of a glutamate (NMDA) subunit receptor gene (GRIN2B) with obsessive-compulsive disorder: a preliminary study. Psychopharmacology (Berl) 2004;174:530–538.
- Laje G, Paddock S, Manji H, Rush AJ, Wilson AF, Charney D, McMahon FJ: Genetic markers of suicidal ideation emerging during citalopram treatment of major depression. Am J Psychiatry 2007;164:1530–1538.
- Need AC, Keefe RS, Ge D, Grossman I, Dickson S, McEvoy JP, Goldstein DB: Pharmacogenetics of antipsychotic response in the CATIE trial: a candidate gene analysis. Eur J Hum Genet 2009;17:946–957.
- Ronnback L, Hansson E: On the potential role of glutamate transport in mental fatigue. J Neuroinflammation 2004;1:22.
- Barnard AR, Nolan PM: When clocks go bad: neurobehavioural consequences of disrupted circadian timing. PLoS Genet 2008;4:e1000040.
- Wulff K, Porcheret K, Cussans E, Foster RG: Sleep and circadian rhythm disturbances: multiple genes and multiple phenotypes. Curr Opin Genet Dev 2009;19:237–246.
- DeBruyne JP, Weaver DR, Reppert SM: CLOCK and NPAS2 have overlapping roles in the suprachiasmatic circadian clock. Nat Neurosci 2007;10:543–545.
- Reick M, Garcia JA, Dudley C, McKnight SL: NPAS2: an analog of clock operative in the mammalian forebrain. Science 2001;293:506–509.
- Garcia JA, Zhang D, Estill SJ, Michnoff C, Rutter J, Reick M, Scott K, Diaz-Arrastia R, McKnight SL: Impaired cued and contextual memory in NPAS2-deficient mice. Science 2000;288:2226–2230.
- Franken P, Dudley CA, Estill SJ, Barakat M, Thomason R, O’Hara BF, McKnight SL: NPAS2 as a transcriptional regulator of non-rapid eye movement sleep: genotype and sex interactions. Proc Natl Acad Sci USA 2006;103:7118–7123.
- Wisor JP, Pasumarthi RK, Gerashchenko D, Thompson CL, Pathak S, Sancar A, Franken P, Lein ES, Kilduff TS: Sleep deprivation effects on circadian clock gene expression in the cerebral cortex parallel electroencephalographic differences among mouse strains. J Neurosci 2008;28:7193–7201.
- Nicholas B, Rudrasingham V, Nash S, Kirov G, Owen MJ, Wimpory DC: Association of Per1 and Npas2 with autistic disorder: support for the clock genes/social timing hypothesis. Mol Psychiatry 2007;12:581–592.
- Partonen T, Treutlein J, Alpman A, Frank J, Johansson C, Depner M, Aron L, Rietschel M, Wellek S, Soronen P, Paunio T, Koch A, Chen P, Lathrop M, Adolfsson R, Persson ML, Kasper S, Schalling M, Peltonen L, Schumann G: Three circadian clock genes Per2, Arntl, and Npas2 contribute to winter depression. Ann Med 2007;39:229–238.
- Mansour HA, Talkowski ME, Wood J, Chowdari KV, McClain L, Prasad K, Montrose D, Fagiolini A, Friedman ES, Allen MH, Bowden CL, Calabrese J, El-Mallakh RS, Escamilla M, Faraone SV, Fossey MD, Gyulai L, Loftis JM, Hauser P, Ketter TA, Marangell LB, Miklowitz DJ, Nierenberg AA, Patel J, Sachs GS, Sklar P, Smoller JW, Laird N, Keshavan M, Thase ME, Axelson D, Birmaher B, Lewis D, Monk T, Frank E, Kupfer DJ, Devlin B, Nimgaonkar VL: Association study of 21 circadian genes with bipolar I disorder, schizoaffective disorder, and schizophrenia. Bipolar Disord 2009;11:701–710.
- Soria V, Martínez-Amorós E, Escaramís G, Valero J, Pérez-Egea R, García C, Gutiérrez-Zotes A, Puigdemont D, Bayés M, Crespo JM, Martorell L, Vilella E, Labad A, Vallejo J, Pérez V, Menchón JM, Estivill X, Gratacòs M, Urretavizcaya M: Differential association of circadian genes with mood disorders: CRY1 and NPAS2 are associated with unipolar major depression and CLOCK and VIP with bipolar disorder. Neuropsychopharmacology 2010;35:1279–1289.
- Hoffman AE, Zheng T, Ba Y, Zhu Y: The circadian gene NPAS2, a putative tumor suppressor, is involved in DNA damage response. Mol Cancer Res 2008;6:1461–1468.
- Levine PH, Peterson D, McNamee FL, O’Brien K, Gridley G, Hagerty M, Brady J, Fears T, Atherton M, Hoover R: Does chronic fatigue syndrome predispose to non-Hodgkin’s lymphoma? Cancer Res 1992;52(19 suppl):5516s–5518s, discussion 5518s–5521s.
- Bass J, Takahashi JS: Circadian integration of metabolism and energetics. Science 2010;330:1349–1354.
- Yu M, Riva L, Xie H, Schindler Y, Moran TB, Cheng Y, Yu D, Hardison R, Weiss MJ, Orkin SH, Bernstein BE, Fraenkel E, Cantor AB: Insights into GATA-1-mediated gene activation versus repression via genome-wide chromatin occupancy analysis. Mol Cell 2009;36:682–695.
- Zhao M, Sun Y, Gao F, Wu X, Tang J, Yin H, Luo Y, Richardson B, Lu Q: Epigenetics and SLE: RFX1 downregulation causes CD11a and CD70 overexpression by altering epigenetic modifications in lupus CD4+ T cells. J Autoimmun 2010;35:58–69.
- Whistler T, Fletcher MA, Lonergan W, Zeng XR, Lin JM, Laperriere A, Vernon SD, Klimas NG: Impaired immune function in Gulf War Illness. BMC Med Genomics 2009;2:12.
- Hickie I, Davenport T, Wakefield D, Vollmer-Conna U, Cameron B, Vernon SD, Reeves WC, Lloyd A; Dubbo Infection Outcomes Study Group: Post-infective and chronic fatigue syndromes precipitated by viral and non-viral pathogens: prospective cohort study. BMJ 2006;333:575.
- Vernon SD, Nicholson A, Rajeevan M, Dimulescu I, Cameron B, Whistler T, Lloyd A: Correlation of psycho-neuroendocrine-immune (PNI) gene expression with symptoms of acute infectious mononucleosis. Brain Res 2006;1068:1–6.
- Sikder D, Kodadek T: The neurohormone orexin stimulates hypoxia-inducible factor-1 activity. Genes Dev 2007;21:2995–3005.
- Silverman MN, Pearce BD, Biron CA, Miller AH: Immune modulation of the hypothalamic-pituitary-adrenal (HPA) axis during viral infection. Viral Immunol 2005;18:41–78.
- Nicholson AC, Unger ER, Mangalathu R, Ojaniemi H, Vernon SD: Exploration of neuroendocrine and immune gene expression in peripheral blood mononuclear cells. Brain Res Mol Brain Res 2004;129:193–197.
Mangalathu S. Rajeevan
Division of High-Consequence Pathogens and Pathology
Centers for Disease Control and Prevention, 1600 Clifton Road
Atlanta, GA 30333 (USA)
Tel. +1 404 639 2931, E-Mail email@example.com
The findings and conclusions in this report are those of the authors and do not necessarily represent the views of the funding agency.
Received: August 26, 2010
Accepted after revision: February 21, 2011
Published online: September 9, 2011
Number of Print Pages : 12
Number of Figures : 0, Number of Tables : 2, Number of References : 60
Additional supplementary material is available online - Number of Parts : 3
Neuropsychobiology (International Journal of Experimental and Clinical Research in Biological Psychiatry, Pharmacopsychiatry, Biological Psychology/Pharmacopsychology and Pharmacoelectroencephalography)
Vol. 64, No. 4, Year 2011 (Cover Date: September 2011)
Journal Editor: Strik W. (Bern)
ISSN: 0302-282X (Print), eISSN: 1423-0224 (Online)
For additional information: http://www.karger.com/NPS
Copyright / Drug Dosage / Disclaimer
Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher or, in the case of photocopying, direct payment of a specified fee to the Copyright Clearance Center.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in goverment regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.