Assessing the metabolic effects of prednisolone in healthy volunteers using urine metabolic profiling

Sandrine Ellero-Simatos, Ewa Szymańska, Ton Rullmann, Wim Ha Dokter, Raymond Ramaker, Ruud Berger, Thijs Mp van Iersel, Age K Smilde, Thomas Hankemeier, Wynand Alkema, Sandrine Ellero-Simatos, Ewa Szymańska, Ton Rullmann, Wim Ha Dokter, Raymond Ramaker, Ruud Berger, Thijs Mp van Iersel, Age K Smilde, Thomas Hankemeier, Wynand Alkema

Abstract

Background: Glucocorticoids, such as prednisolone, are widely used anti-inflammatory drugs, but therapy is hampered by a broad range of metabolic side effects including skeletal muscle wasting and insulin resistance. Therefore, development of improved synthetic glucocorticoids that display similar efficacy as prednisolone but reduced side effects is an active research area. For efficient development of such new drugs, in vivo biomarkers, which can predict glucocorticoid metabolic side effects in an early stage, are needed. In this study, we aim to provide the first description of the metabolic perturbations induced by acute and therapeutic treatments with prednisolone in humans using urine metabolomics, and to derive potential biomarkers for prednisolone-induced metabolic effects.

Methods: A randomized, double blind, placebo-controlled trial consisting of two protocols was conducted in healthy men. In protocol 1, volunteers received placebo (n = 11) or prednisolone (7.5 mg (n = 11), 15 mg (n = 13) or 30 mg (n = 12)) orally once daily for 15 days. In protocol 2, volunteers (n = 6) received placebo at day 0 and 75 mg prednisolone at day 1. We collected 24 h urine and serum samples at baseline (day 0), after a single dose (day 1) and after prolonged treatment (day 15) and obtained mass-spectrometry-based urine and serum metabolic profiles.

Results: At day 1, high-dose prednisolone treatment increased levels of 13 and 10 proteinogenic amino acids in urine and serum respectively, as well as levels of 3-methylhistidine, providing evidence for an early manifestation of glucocorticoid-induced muscle wasting. Prednisolone treatment also strongly increased urinary carnitine derivatives at day 1 but not at day 15, which might reflect adaptive mechanisms under prolonged treatment. Finally, urinary levels of proteinogenic amino acids at day 1 and of N-methylnicotinamide at day 15 significantly correlated with the homeostatic model assessment of insulin resistance and might represent biomarkers for prednisolone-induced insulin resistance.

Conclusion: This study provides evidence that urinary metabolomics represents a noninvasive way of monitoring the effect of glucocorticoids on muscle protein catabolism after a single dose and can derive new biomarkers of glucocorticoid-induced insulin resistance. It might, therefore, help the development of improved synthetic glucocorticoids.

Trial registration: ClinicalTrials.gov NCT00971724.

Keywords: 3-methylhistidine; HOMA-IR; aminoaciduria; metabolomics; prednisolone; urine.

Figures

Figure 1
Figure 1
PCA plots of urinary metabolic profiles. (A) The first PCA model includes metabolic profiles from block 1 volunteers treated with placebo (black, n = 11) or 30 mg prednisolone (dark red, n = 12) for one day (circle) or 15 days (square). (B) The second PCA model includes metabolic profiles from block 1 volunteers treated with placebo (black, n = 11) or 7.5 mg (orange, n = 11), 15 mg (pink, n = 13) or 30 mg (dark red, n = 12) prednisolone for one day. (C) The third PCA model includes metabolic profiles from block 1 volunteers treated with placebo or prednisolone for 15 days. Arrows represent dose-dependent metabolic trajectories.
Figure 2
Figure 2
3-methylhistidine in protocol 2 volunteers. Data represent metabolite levels (divided by the mean of 3-methylhistidine level detected in this study) in urine (A) and serum (B) of protocol 2 volunteers before and after a single dose of prednisolone (75 mg). P-values calculated using paired t tests.
Figure 3
Figure 3
HOMA-IR n volunteers from protocol 1. (A) Day 2. (B) Day 16. The black lines represent the mean value. The top and bottom of the box represent the 75th and 25th percentile. The whiskers indicate the maximum and minimum points. *P <0.05 compared to placebo group, using analysis of variance.

References

    1. Schacke H, Docke WD, Asadullah K. Mechanisms involved in the side effects of glucocorticoids. Pharmacol Ther. 2002;4:23–43. doi: 10.1016/S0163-7258(02)00297-8.
    1. van Raalte DH, Ouwens DM, Diamant M. Novel insights into glucocorticoid-mediated diabetogenic effects: towards expansion of therapeutic options?. Eur J Clin Invest. 2009;4:81–93. doi: 10.1111/j.1365-2362.2008.02067.x.
    1. Jacobson PB, von Geldern TW, Ohman L, Osterland M, Wang J, Zinker B, Wilcox D, Nguyen PT, Mika A, Fung S. et al.Hepatic glucocorticoid receptor antagonism is sufficient to reduce elevated hepatic glucose output and improve glucose control in animal models of type 2 diabetes. The Journal of pharmacology and experimental therapeutics. 2005;4:191–200. doi: 10.1124/jpet.104.081257.
    1. Macfarlane DP, Forbes S, Walker BR. Glucocorticoids and fatty acid metabolism in humans: fuelling fat redistribution in the metabolic syndrome. J Endocrinol. 2008;4:189–204. doi: 10.1677/JOE-08-0054.
    1. Schacke H, Schottelius A, Docke WD, Strehlke P, Jaroch S, Schmees N, Rehwinkel H, Hennekes H, Asadullah K. Dissociation of transactivation from transrepression by a selective glucocorticoid receptor agonist leads to separation of therapeutic effects from side effects. Proc Natl Acad Sci USA. 2004;4:227–232. doi: 10.1073/pnas.0300372101.
    1. Barnes PJ. Anti-inflammatory actions of glucocorticoids: molecular mechanisms. Clin Sci (Lond) 1998;4:557–572.
    1. Miner JN, Ardecky B, Benbatoul K, Griffiths K, Larson CJ, Mais DE, Marschke K, Rosen J, Vajda E, Zhi L, Negro-Vilar A. Antiinflammatory glucocorticoid receptor ligand with reduced side effects exhibits an altered protein-protein interaction profile. Proc Natl Acad Sci USA. 2007;4:19244–19249. doi: 10.1073/pnas.0705517104.
    1. Wishart DS. Applications of metabolomics in drug discovery and development. Drugs R D. 2008;4:307–322. doi: 10.2165/00126839-200809050-00002.
    1. Beger RD, Sun J, Schnackenberg LK. Metabolomics approaches for discovering biomarkers of drug-induced hepatotoxicity and nephrotoxicity. Toxicol Appl Pharmacol. 2010;4:154–166. doi: 10.1016/j.taap.2009.11.019.
    1. Clayton TA, Baker D, Lindon JC, Everett JR, Nicholson JK. Pharmacometabonomic identification of a significant host-microbiome metabolic interaction affecting human drug metabolism. Proc Natl Acad Sci USA. 2009;4:14728–14733. doi: 10.1073/pnas.0904489106.
    1. van Raalte DH, Nofrate V, Bunck MC, van Iersel T, Elassaiss Schaap J, Nassander UK, Heine RJ, Mari A, Dokter WH, Diamant M. Acute and 2-week exposure to prednisolone impair different aspects of beta-cell function in healthy men. Eur J Endocrinol. 2010;4:729–735. doi: 10.1530/EJE-09-1034.
    1. Lawton KA, Berger A, Mitchell M, Milgram KE, Evans AM, Guo L, Hanson RW, Kalhan SC, Ryals JA, Milburn MV. Analysis of the adult human plasma metabolome. Pharmacogenomics. 2008;4:383–397. doi: 10.2217/14622416.9.4.383.
    1. Boudonck KJ, Mitchell MW, Nemet L, Keresztes L, Nyska A, Shinar D, Rosenstock M. Discovery of metabolomics biomarkers for early detection of nephrotoxicity. Toxicol Pathol. 2009;4:280–292. doi: 10.1177/0192623309332992.
    1. Noga MJ, Dane A, Shi S, Attali A, van Aken H, Suidgeest E, Tuinstra T, Muilwijk B, Coulier L, Luider T. et al.Metabolomics of cerebrospinal fluid reveals change in the central nervous system metabolism in a rat model of multiple sclerosis. Metabolomics. 2012;4:253–263. doi: 10.1007/s11306-011-0306-3.
    1. R: A Language and Environment for Statistical Computing.
    1. Storey JD, Tibshirani R. Statistical significance for genomewide studies. Proc Natl Acad Sci USA. 2003;4:9440–9445. doi: 10.1073/pnas.1530509100.
    1. Szymanska E, vDF A, Troost J, Paliukhovich I, van Velzen EJJ, Hendriks MMWB, Trautwein EA, van Duynhoven JPM, Vreeken RJ, Smilde AK. A lipidomic analysis approach to evaluate the response to cholesterol-lowering food intake. Metabolomics. 2011.
    1. Short KR, Bigelow ML, Nair KS. Short-term prednisone use antagonizes insulin's anabolic effect on muscle protein and glucose metabolism in young healthy people. Am J Physiol-Endoc M. 2009;4:E1260–E1268.
    1. Gravholt CH, Dall R, Christiansen JS, Moller N, Schmitz O. Preferential stimulation of abdominal subcutaneous lipolysis after prednisolone exposure in humans. Obes Res. 2002;4:774–781. doi: 10.1038/oby.2002.105.
    1. van Raalte DH, Brands M, van der Zijl NJ, Muskiet MH, Pouwels PJ, Ackermans MT, Sauerwein HP, Serlie MJ, Diamant M. Low-dose glucocorticoid treatment affects multiple aspects of intermediary metabolism in healthy humans: a randomised controlled trial. Diabetologia. 2011;4:2103–2112. doi: 10.1007/s00125-011-2174-9.
    1. Silbernagl S. The renal handling of amino acids and oligopeptides. Physiol Rev. 1988;4:911–1007.
    1. Fleck C, Schwertfeger M, Taylor PM. Regulation of renal amino acid (AA) transport by hormones, drugs and xenobiotics - a review. Amino Acids. 2003;4:347–374. doi: 10.1007/s00726-002-0316-6.
    1. Schwertfeger M, Pissowotzki K, Fleck C, Taylor PM. Regulation of L-leucine transport in rat kidney by dexamethasone and triiodothyronine. Amino Acids. 2003;4:75–83.
    1. Beaufrere B, Horber FF, Schwenk WF, Marsh HM, Matthews D, Gerich JE, Haymond MW. Glucocorticosteroids increase leucine oxidation and impair leucine balance in humans. Am J Physiol. 1989;4:E712–721.
    1. Horber FF, Haymond MW. Human growth hormone prevents the protein catabolic side effects of prednisone in humans. J Clin Invest. 1990;4:265–272. doi: 10.1172/JCI114694.
    1. Garrel DR, Moussali R, De Oliveira A, Lesiege D, Lariviere F. RU 486 prevents the acute effects of cortisol on glucose and leucine metabolism. J Clin Endocrinol Metab. 1995;4:379–385. doi: 10.1210/jc.80.2.379.
    1. Burt MG, Johannsson G, Umpleby AM, Chisholm DJ, Ho KK. Impact of acute and chronic low-dose glucocorticoids on protein metabolism. J Clin Endocrinol Metab. 2007;4:3923–3929. doi: 10.1210/jc.2007-0951.
    1. Schakman O, Gilson H, Thissen JP. Mechanisms of glucocorticoid-induced myopathy. J Endocrinol. 2008;4:1–10. doi: 10.1677/JOE-07-0606.
    1. Tomas FM, Munro HN, Young VR. Effect of glucocorticoid administration on the rate of muscle protein breakdown in vivo in rats, as measured by urinary excretion of N tau-methylhistidine. Biochem J. 1979;4:139–146.
    1. Lofberg E, Gutierrez A, Wernerman J, Anderstam B, Mitch WE, Price SR, Bergstrom J, Alvestrand A. Effects of high doses of glucocorticoids on free amino acids, ribosomes and protein turnover in human muscle. Eur J Clin Invest. 2002;4:345–353. doi: 10.1046/j.1365-2362.2002.00993.x.
    1. Young VR, Alexis SD, Baliga BS, Munro HN, Muecke W. Metabolism of administered 3-methylhistidine. Lack of muscle transfer ribonucleic acid charging and quantitative excretion as 3-methylhistidine and its N-acetyl derivative. The Journal of biological chemistry. 1972;4:3592–3600.
    1. Young VR, Havenberg LN, Bilmazes C, Munro HN. Potential use of 3-methylhistidine excretion as an index of progressive reduction in muscle protein catabolism during starvation. Metabolism. 1973;4:1429–1436.
    1. Afting EG, Bernhardt W, Janzen RW, Rothig HJ. Quantitative importance of non-skeletal-muscle N tau-methylhistidine and creatine in human urine. Biochem J. 1981;4:449–452.
    1. Rennie MJ, Bennegard K, Eden E, Emery PW, Lundholm K. Urinary excretion and efflux from the leg of 3-methylhistidine before and after major surgical operation. Metabolism. 1984;4:250–256. doi: 10.1016/0026-0495(84)90046-5.
    1. Millward DJ, Bates PC, Grimble GK, Brown JG, Nathan M, Rennie MJ. Quantitative importance of non-skeletal-muscle sources of N tau-methylhistidine in urine. Biochem J. 1980;4:225–228.
    1. Emery PW, Cotellessa L, Holness M, Egan C, Rennie MJ. Different patterns of protein turnover in skeletal and gastrointestinal smooth muscle and the production of N tau-methylhistidine during fasting in the rat. Bioscience reports. 1986;4:143–153. doi: 10.1007/BF01115000.
    1. Trappe T, Williams R, Carrithers J, Raue U, Esmarck B, Kjaer M, Hickner R. Influence of age and resistance exercise on human skeletal muscle proteolysis: a microdialysis approach. The Journal of physiology. 2004;4:803–813.
    1. Pereira RM, Freire de Carvalho J. Glucocorticoid-induced myopathy. Joint Bone Spine. 2011;4:41–44. doi: 10.1016/j.jbspin.2010.02.025.
    1. Rebouche CJ, Engel AG. Kinetic compartmental analysis of carnitine metabolism in the human carnitine deficiency syndromes. Evidence for alterations in tissue carnitine transport. J Clin Invest. 1984;4:857–867. doi: 10.1172/JCI111281.
    1. Patterson AD, Slanar O, Krausz KW, Li F, Hofer CC, Perlik F, Gonzalez FJ, Idle JR. Human urinary metabolomic profile of PPARalpha induced fatty acid beta-oxidation. J Proteome Res. 2009;4:4293–4300. doi: 10.1021/pr9004103.
    1. Rittmaster RS, Loriaux DL, Cutler GB Jr. Sensitivity of cortisol and adrenal androgens to dexamethasone suppression in hirsute women. J Clin Endocrinol Metab. 1985;4:462–466. doi: 10.1210/jcem-61-3-462.
    1. Valenti G. Adrenopause: an imbalance between dehydroepiandrosterone (DHEA) and cortisol secretion. J Endocrinol Invest. 2002;4:29–35.
    1. Valenti G. Neuroendocrine hypothesis of aging: the role of corticoadrenal steroids. J Endocrinol Invest. 2004;4:62–63.
    1. Arlt W. Dehydroepiandrosterone replacement therapy. Semin Reprod Med. 2004;4:379–388. doi: 10.1055/s-2004-861554.
    1. Tai ES, Tan ML, Stevens RD, Low YL, Muehlbauer MJ, Goh DL, Ilkayeva OR, Wenner BR, Bain JR, Lee JJ. et al.Insulin resistance is associated with a metabolic profile of altered protein metabolism in Chinese and Asian-Indian men. Diabetologia. 2010;4:757–767. doi: 10.1007/s00125-009-1637-8.
    1. Salek RM, Maguire ML, Bentley E, Rubtsov DV, Hough T, Cheeseman M, Nunez D, Sweatman BC, Haselden JN, Cox RD. et al.A metabolomic comparison of urinary changes in type 2 diabetes in mouse, rat, and human. Physiol Genomics. 2007;4:99–108.
    1. Thomas MC, Jerums G, Tsalamandris C, Macisaac R, Panagiotopoulos S, Cooper ME. Increased tubular organic ion clearance following chronic ACE inhibition in patients with type 1 diabetes. Kidney Int. 2005;4:2494–2499. doi: 10.1111/j.1523-1755.2005.00359.x.
    1. Zhou SS, Li D, Sun WP, Guo M, Lun YZ, Zhou YM, Xiao FC, Jing LX, Sun SX, Zhang LB. et al.Nicotinamide overload may play a role in the development of type 2 diabetes. World J Gastroenterol. 2009;4:5674–5684. doi: 10.3748/wjg.15.5674.
    1. Chang AM, Smith MJ, Galecki AT, Bloem CJ, Halter JB. Impaired beta-cell function in human aging: response to nicotinic acid-induced insulin resistance. J Clin Endocrinol Metab. 2006;4:3303–3309. doi: 10.1210/jc.2006-0913.
    1. Kahn SE, Beard JC, Schwartz MW, Ward WK, Ding HL, Bergman RN, Taborsky GJ Jr, Porte D Jr. Increased beta-cell secretory capacity as mechanism for islet adaptation to nicotinic acid-induced insulin resistance. Diabetes. 1989;4:562–568. doi: 10.2337/diabetes.38.5.562.

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