Effect of short-term oral prednisone therapy on blood gene expression: a randomised controlled clinical trial

Hiroto Takiguchi, Virginia Chen, Ma'en Obeidat, Zsuzsanna Hollander, J Mark FitzGerald, Bruce M McManus, Raymond T Ng, Don D Sin, Hiroto Takiguchi, Virginia Chen, Ma'en Obeidat, Zsuzsanna Hollander, J Mark FitzGerald, Bruce M McManus, Raymond T Ng, Don D Sin

Abstract

Background: Effects of systemic corticosteroids on blood gene expression are largely unknown. This study determined gene expression signature associated with short-term oral prednisone therapy in patients with chronic obstructive pulmonary disease (COPD) and its relationship to 1-year mortality following an acute exacerbation of COPD (AECOPD).

Methods: Gene expression in whole blood was profiled using the Affymetrix Human Gene 1.1 ST microarray chips from two cohorts: 1) a prednisone cohort with 37 stable COPD patients randomly assigned to prednisone 30 mg/d + standard therapy for 4 days or standard therapy alone and 2) the Rapid Transition Program (RTP) cohort with 218 COPD patients who experienced AECOPD and were treated with systemic corticosteroids. All gene expression data were adjusted for the total number of white blood cells and their differential cell counts.

Results: In the prednisone cohort, 51 genes were differentially expressed between prednisone and standard therapy group at a false discovery rate of < 0.05. The top 3 genes with the largest fold-changes were KLRF1, GZMH and ADGRG1; and 21 genes were significantly enriched in immune system pathways including the natural killer cell mediated cytotoxicity. In the RTP cohort, 27 patients (12.4%) died within 1 year after hospitalisation of AECOPD; 32 of 51 genes differentially expressed in the prednisone cohort significantly changed from AECOPD to the convalescent state and were enriched in similar cellular immune pathways to that in the prednisone cohort. Of these, 10 genes including CX3CR1, KLRD1, S1PR5 and PRF1 were significantly associated with 1-year mortality.

Conclusions: Short-term daily prednisone therapy produces a distinct blood gene signature that may be used to determine and monitor treatment responses to prednisone in COPD patients during AECOPD.

Trial registration: The prednisone cohort was registered at clinicalTrials.gov ( NCT02534402 ) and the RTP cohort was registered at ClinicalTrials.gov ( NCT02050022 ).

Keywords: Acute exacerbation; Blood; Chronic obstructive pulmonary disease; Gene expression; Microarray; Mortality; Prednisone.

Conflict of interest statement

The authors declare that they have no competing interests.

Figures

Fig. 1
Fig. 1
A flow diagram of participants. In study 1, 40 patients were enrolled and were randomised to either prednisone (30 mg/d for 4 days) or control group in a 3: 1 ratio. Participants attended 5 study visits over 5 consecutive days; 28 and 9 patients were included in the final analysis after excluding 2 and 1 patients from prednisone and control groups, respectively
Fig. 2
Fig. 2
A Volcano plot of gene differentially expressed between prednisone and control group. The plot shows the fold-change on the X-axis versus the unadjusted P values (on the–log10 scale) on the Y-axis. Genes at a FDR < 0.05 for differential expression in study 1 are annotated on the graph (coloured dots). The red dots represent replicated genes in study 2 and genes that related to 1-year mortality are represented as dark red dots
Fig. 3
Fig. 3
Theoretical survival curves based on gene signature of replicated 10 genes. 90th versus 10th risk percentile of 10 genes performed by fitting Cox proportional-hazards was shown. Broken lines represent the 95% confidence interval

References

    1. Mokdad AH, Ballestros K, Echko M, Glenn S, Olsen HE, Mullany E, Lee A, Khan AR, Ahmadi A, Ferrari AJ, et al. The state of US health, 1990-2016: burden of diseases, injuries, and risk factors among US states. Jama. 2018;319:1444–1472. doi: 10.1001/jama.2018.0158.
    1. COPD Hospitalizations . Accessed 20 Jan 2019.
    1. Quon BS, Gan WQ, Sin DD. Contemporary management of acute exacerbations of COPD: a systematic review and metaanalysis. Chest. 2008;133:756–766. doi: 10.1378/chest.07-1207.
    1. Leuppi JD, Schuetz P, Bingisser R, Bodmer M, Briel M, Drescher T, Duerring U, Henzen C, Leibbrandt Y, Maier S, et al. Short-term vs conventional glucocorticoid therapy in acute exacerbations of chronic obstructive pulmonary disease: the REDUCE randomized clinical trial. Jama. 2013;309:2223–2231. doi: 10.1001/jama.2013.5023.
    1. García-Sanz María-Teresa, Cánive-Gómez Juan-Carlos, Senín-Rial Laura, Aboal-Viñas Jorge, Barreiro-García Alejandra, López-Val Eva, González-Barcala Francisco-Javier. One-year and long-term mortality in patients hospitalized for chronic obstructive pulmonary disease. Journal of Thoracic Disease. 2017;9(3):636–645. doi: 10.21037/jtd.2017.03.34.
    1. Ho Te-Wei, Tsai Yi-Ju, Ruan Sheng-Yuan, Huang Chun-Ta, Lai Feipei, Yu Chong-Jen. In-Hospital and One-Year Mortality and Their Predictors in Patients Hospitalized for First-Ever Chronic Obstructive Pulmonary Disease Exacerbations: A Nationwide Population-Based Study. PLoS ONE. 2014;9(12):e114866. doi: 10.1371/journal.pone.0114866.
    1. Leung JM, Chen V, Hollander Z, Dai D, Tebbutt SJ, Aaron SD, Vandemheen KL, Rennard SI, FitzGerald JM, Woodruff PG, et al. COPD exacerbation biomarkers validated using multiple reaction monitoring mass spectrometry. PLoS One. 2016;11:e0161129. doi: 10.1371/journal.pone.0161129.
    1. Wang J, Vasaikar S, Shi Z, Greer M, Zhang B. WebGestalt 2017: a more comprehensive, powerful, flexible and interactive gene set enrichment analysis toolkit. Nucleic Acids Res. 2017;45:W130–WW37. doi: 10.1093/nar/gkx356.
    1. Ashburner M, Ball CA, Blake JA, Botstein D, Butler H, Cherry JM, Davis AP, Dolinski K, Dwight SS, Eppig JT, et al. Gene ontology: tool for the unification of biology. The Gene Ontology Consortium. Nat Genet. 2000;25:25–29. doi: 10.1038/75556.
    1. Kanehisa M, Goto S, Kawashima S, Okuno Y, Hattori M. The KEGG resource for deciphering the genome. Nucleic Acids Res. 2004;32:D277–D280. doi: 10.1093/nar/gkh063.
    1. Di Stefano A, Caramori G, Oates T, Capelli A, Lusuardi M, Gnemmi I, Ioli F, Chung KF, Donner CF, Barnes PJ, Adcock IM. Increased expression of nuclear factor-kappaB in bronchial biopsies from smokers and patients with COPD. Eur Respir J. 2002;20:556–563. doi: 10.1183/09031936.02.00272002.
    1. Barnes PJ. Corticosteroid effects on cell signalling. Eur Respir J. 2006;27:413–426. doi: 10.1183/09031936.06.00125404.
    1. Barnes PJ, Karin M. Nuclear factor-kappaB: a pivotal transcription factor in chronic inflammatory diseases. N Engl J Med. 1997;336:1066–1071. doi: 10.1056/NEJM199704103361506.
    1. Chang GW, Hsiao CC, Peng YM, Vieira Braga FA, Kragten NA, Remmerswaal EB, van de Garde MD, Straussberg R, Konig GM, Kostenis E, et al. The adhesion G protein-coupled receptor GPR56/ADGRG1 is an inhibitory receptor on human NK cells. Cell Rep. 2016;15:1757–1770. doi: 10.1016/j.celrep.2016.04.053.
    1. Crooks SW, Bayley DL, Hill SL, Stockley RA. Bronchial inflammation in acute bacterial exacerbations of chronic bronchitis: the role of leukotriene B4. Eur Respir J. 2000;15:274–280. doi: 10.1034/j.1399-3003.2000.15b09.x.
    1. Guma M, Busch LK, Salazar-Fontana LI, Bellosillo B, Morte C, Garcia P, Lopez-Botet M. The CD94/NKG2C killer lectin-like receptor constitutes an alternative activation pathway for a subset of CD8+ T cells. Eur J Immunol. 2005;35:2071–2080. doi: 10.1002/eji.200425843.
    1. Kuttruff S, Koch S, Kelp A, Pawelec G, Rammensee HG, Steinle A. NKp80 defines and stimulates a reactive subset of CD8 T cells. Blood. 2009;113:358–369. doi: 10.1182/blood-2008-03-145615.
    1. Lanier LL. Up on the tightrope: natural killer cell activation and inhibition. Nat Immunol. 2008;9:495–502. doi: 10.1038/ni1581.
    1. Middleton D, Curran M, Maxwell L. Natural killer cells and their receptors. Transpl Immunol. 2002;10:147–164. doi: 10.1016/S0966-3274(02)00062-X.
    1. Parham P, Abi-Rached L, Matevosyan L, Moesta AK, Norman PJ, Older Aguilar AM, Guethlein LA. Primate-specific regulation of natural killer cells. J Med Primatol. 2010;39:194–212. doi: 10.1111/j.1600-0684.2010.00432.x.
    1. Barry M, Bleackley RC. Cytotoxic T lymphocytes: all roads lead to death. Nat Rev Immunol. 2002;2:401–409. doi: 10.1038/nri819.
    1. Ngan DA, Vickerman SV, Granville DJ, Man SF, Sin DD. The possible role of granzyme B in the pathogenesis of chronic obstructive pulmonary disease. Ther Adv Respir Dis. 2009;3:113–129. doi: 10.1177/1753465809341965.
    1. Alberts B, Johnson A, Lewis J, Raff M, Roberts K, Walter P. Molecular biology of the cell. 4. New York: Garland Science; 2002. Innate immunity; p. 1616.
    1. Barnawi J, Jersmann H, Haberberger R, Hodge S, Meech R. Reduced DNA methylation of sphingosine-1 phosphate receptor 5 in alveolar macrophages in COPD: a potential link to failed efferocytosis. Respirology. 2017;22:315–321. doi: 10.1111/resp.12949.
    1. McComb JG, Ranganathan M, Liu XH, Pilewski JM, Ray P, Watkins SC, Choi AM, Lee JS. CX3CL1 up-regulation is associated with recruitment of CX3CR1+ mononuclear phagocytes and T lymphocytes in the lungs during cigarette smoke-induced emphysema. Am J Pathol. 2008;173:949–961. doi: 10.2353/ajpath.2008.071034.
    1. Sanchez T, Hla T. Structural and functional characteristics of S1P receptors. J Cell Biochem. 2004;92:913–922. doi: 10.1002/jcb.20127.
    1. Barnawi J, Tran H, Jersmann H, Pitson S, Roscioli E, Hodge G, Meech R, Haberberger R, Hodge S. Potential link between the Sphingosine-1-phosphate (S1P) system and defective alveolar macrophage phagocytic function in chronic obstructive pulmonary disease (COPD) PLoS One. 2015;10:e0122771. doi: 10.1371/journal.pone.0122771.
    1. Burge PS, Calverley PM, Jones PW, Spencer S, Anderson JA. Prednisolone response in patients with chronic obstructive pulmonary disease: results from the ISOLDE study. Thorax. 2003;58:654–658. doi: 10.1136/thorax.58.8.654.
    1. Bhatt Surya P., Anderson Julie A., Brook Robert D., Calverley Peter M.A., Celli Bartolome R., Cowans Nicholas J., Crim Courtney, Martinez Fernando J., Newby David E., Vestbo Jørgen, Yates Julie C., Dransfield Mark T. Cigarette smoking and response to inhaled corticosteroids in COPD. European Respiratory Journal. 2018;51(1):1701393. doi: 10.1183/13993003.01393-2017.

Source: PubMed

Sponsors en medewerkers

Medische omstandigheden

Geneesmiddelinterventies

3
Abonneren