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Poor Inter-test Reliability Between CK18 Kits as a Biomarker of NASH

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Abstract

Background and Aim

Nonalcoholic fatty liver disease (NAFLD) affects 15–40 % of the general population; 10–20 % of those patients have a more severe form of the disease known as nonalcoholic steatohepatitis (NASH). Cytokeratin-18 (CK18), released during apoptosis and one of the most studied biomarkers in NASH, can be measured by a number of commercially available kits. We compared serum measurements of the CK18 M30 from two different kits using the same cohort to evaluate the reliability between two test kits.

Methods

We measured serum levels of CK18 M30 from 185 patients with biopsy-proven NAFLD from a single center from 2009 to 2015, using two different ELISA kits, Test 1 (T1) and Test 2 (T2). Advanced fibrosis was defined as fibrosis stages 3–4 and NASH defined by NAS score ≥5.

Results

Mean age was 50.2 years (SD 12.6), 61.1 % male and 87 % White; 49.6 % had NASH and 32.2 % advanced fibrosis. There was no significant correlation between measurements from the two kits (p = 0.86, r = 0.01). While T2 predicted NASH and advanced fibrosis, T1 did not. The area under ROC curve for the prediction of NASH was 0.631 for T2 versus 0.500 for T1.

Conclusions

Measurements from two different CK18 M30 test kits did not correlate with each other. One kit showed statistically significantly higher levels of CK18 M30 in patients with advanced fibrosis and NASH, while the other kit did not. With the increasing use of CK18 as a biomarker in NASH, it is important to standardize the different kits as it could greatly bias the results.

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Abbreviations

NAFLD:

Nonalcoholic liver disease

NASH:

Nonalcoholic steatohepatitis

CK18:

Cytokeratin 18

AUROC:

Area under the receiver operating characteristic curve

NAFL:

Nonalcoholic fatty liver, simple steatosis

HR:

Hazard ratio

References

  1. Vernon G, Baranova A, Younossi ZM. Systematic review: the epidemiology and natural history of non-alcoholic fatty liver disease and non-alcoholic steatohepatitis in adults. Aliment Pharmacol Ther. 2011;34:274–285.

    Article  CAS  PubMed  Google Scholar 

  2. Browning JD, Szczepaniak LS, Dobbins R, et al. Prevalence of hepatic steatosis in an urban population in the United States: impact of ethnicity. Hepatology. 2004;40:1387–1395.

    Article  PubMed  Google Scholar 

  3. Williams CD, Stengel J, Asike MI, et al. Prevalence of nonalcoholic fatty liver disease and nonalcoholic steatohepatitis among a largely middle-aged population utilizing ultrasound and liver biopsy: a prospective study. Gastroenterology. 2011;140:124–131.

    Article  PubMed  Google Scholar 

  4. Younossi ZM, Stepanova M, Afendy M, et al. Changes in the prevalence of the most common causes of chronic liver diseases in the United States From 1988 to 2008. Clin Gastroenterol Hepatol. 2011;9:524–530.

    Article  PubMed  Google Scholar 

  5. Adams LA, Sanderson S, Lindor KD, Angulo P. The histological course of nonalcoholic fatty liver disease: a longitudinal study of 103 patients with sequential liver biopsies. J Hepatol. 2005;42:132–138.

    Article  PubMed  Google Scholar 

  6. Teli MR, James OF, Burt AD, Bennett MK, Day CP. The natural history of nonalcoholic fatty liver: a follow-up study. Hepatology. 1995;22:1714–1719.

    Article  CAS  PubMed  Google Scholar 

  7. Starley BQ, Calcagno CJ, Harrison SA. Nonalcoholic fatty liver disease and hepatocellular carcinoma: a weighty connection. Hepatology. 2010;51:1820–1832.

    Article  PubMed  Google Scholar 

  8. Bugianesi E, Leone N, Vanni E, et al. Expanding the natural history of nonalcoholic steatohepatitis: from cryptogenic cirrhosis to hepatocellular carcinoma. Gastroenterology. 2002;123:134–140.

    Article  PubMed  Google Scholar 

  9. Chalasani N, Younossi Z, Lavine JE, et al. The diagnosis and management of non-alcoholic fatty liver disease: practice guideline by the American Gastroenterological Association, American Association for the Study of Liver Diseases, and American College of Gastroenterology. Gastroenterology. 2012;142:1592–1609.

    Article  PubMed  Google Scholar 

  10. Ong JP, Younossi ZM. Epidemiology and natural history of NAFLD and NASH. Clin Liver Dis. 2007;11:1–16.

    Article  PubMed  Google Scholar 

  11. Singh S, Allen AM, Wang Z, Prokop LJ, Murad MH, Loomba R. Fibrosis progression in nonalcoholic fatty liver vs nonalcoholic steatohepatitis: a systematic review and meta-analysis of paired-biopsy studies. Clin Gastroenterol Hepatol. 2015;13:643.e9–654.e9.

    Google Scholar 

  12. McPherson S, Hardy T, Henderson E, Burt AD, Day CP, Anstee QM. Evidence of NAFLD progression from steatosis to fibrosing-steatohepatitis using paired biopsies: implications for prognosis and clinical management. J Hepatol. 2014;62:1148–1155.

    Article  PubMed  Google Scholar 

  13. Pais R, Charlotte F, Fedchuk L, et al. A systematic review of follow-up biopsies reveals disease progression in patients with non-alcoholic fatty liver. J Hepatol. 2013;59:550–556.

    Article  CAS  PubMed  Google Scholar 

  14. Ekstedt M, Hagström H, Nasr P, et al. Fibrosis stage is the strongest predictor for disease-specific mortality in NAFLD after up to 33 years of follow-up. Hepatology. 2015;61:1547–1554.

    Article  CAS  PubMed  Google Scholar 

  15. Kramer G, Erdal H, Mertens HJMM, et al. Differentiation between cell death modes using measurements of different soluble forms of extracellular cytokeratin 18. Cancer Res. 2004;64:1751–1756.

    Article  CAS  PubMed  Google Scholar 

  16. Wieckowska A, Zein NN, Yerian LM, Lopez AR, McCullough AJ, Feldstein AE. In vivo assessment of liver cell apoptosis as a novel biomarker of disease severity in nonalcoholic fatty liver disease. Hepatology. 2006;44:27–33.

    Article  CAS  PubMed  Google Scholar 

  17. Yilmaz Y. Systematic review: caspase-cleaved fragments of cytokeratin 18—the promises and challenges of a biomarker for chronic liver disease. Aliment Pharmacol Ther. 2009;30:1103–1109.

    Article  CAS  PubMed  Google Scholar 

  18. Aida Y, Abe H, Tomita Y, et al. Serum cytokeratin 18 fragment level as a noninvasive biomarker for non-alcoholic fatty liver disease. Int J Clin Exp Med. 2014;7:4191–4198.

    PubMed Central  PubMed  Google Scholar 

  19. Younossi ZM, Jarrar M, Nugent C, et al. A novel diagnostic biomarker panel for obesity-related nonalcoholic steatohepatitis (NASH). Obes Surg. 2008;18:1430–1437.

    Article  PubMed  Google Scholar 

  20. Cao W, Zhao C, Shen C, Wang Y. Cytokeratin 18, alanine aminotransferase, platelets and triglycerides predict the presence of nonalcoholic steatohepatitis. PLoS One. 2013;8:1–8.

    CAS  Google Scholar 

  21. Anty R, Iannelli A, Patouraux S, Bonnafous S, Lavallard VJ, Amor IB. A new composite model including metabolic syndrome, alanine aminotransferase and cytokeratin-18 for the diagnosis of non-alcoholic steatohepatitis in morbidly obese patients. Aliment Pharmacol Ther. 2010;32:1315–1322.

    Article  CAS  PubMed  Google Scholar 

  22. Feldstein AE, Wieckowska A, Lopez AR, Liu YC, Zein NN, McCullough AJ. Cytokeratin-18 fragment levels as noninvasive biomarkers for nonalcoholic steatohepatitis: a multicenter validation study. Hepatology. 2009;50:1072–1078.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  23. Chan W-K, Sthaneshwar P, Mustapha NRN, Mahadeva S. Limited utility of plasma M30 in discriminating non-alcoholic steatohepatitis from steatosis—a comparison with routine biochemical markers. PLoS One. 2014;9:e105903.

    Article  PubMed Central  PubMed  Google Scholar 

  24. Cusi K, Chang Z, Harrison S, et al. Limited value of plasma cytokeratin-18 as a biomarker for NASH and fibrosis in patients with non-alcoholic fatty liver disease. J Hepatol. 2014;60:167–174.

    Article  CAS  PubMed  Google Scholar 

  25. Kleiner DE, Brunt EM, Van Natta M, et al. Design and validation of a histological scoring system for nonalcoholic fatty liver disease. Hepatology. 2005;41:1313–1321.

    Article  PubMed  Google Scholar 

  26. Yilmaz Y, Dolar E, Ulukaya E, et al. Soluble forms of extracellular cytokeratin 18 may differentiate simple steatosis from nonalcoholic steatohepatitis. World J Gastroenterol. 2007;13:837–844.

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  27. Shen J, Chan HL-Y, Wong GL-H, et al. Assessment of non-alcoholic fatty liver disease using serum total cell death and apoptosis markers. Aliment Pharmacol Ther. 2012;36:1057–1066.

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

This publication was supported by NIDDK Grant 1K23DK083439 to Michelle Lai, MD (PI). The content is solely the responsibility of the authors and does not necessarily represent the official views of the NIDDK or the NIH.

Author contributions

C.F.M.G.P. and M.L. were involved in the primary study design and data analysis; C.F.M.G.P., Z.G.J., T.O., and L.F. were involved in CK18 level measurements; T.L.C. and I.N. were responsible for liver pathology analysis; S.R. and N.A. were responsible for manuscript analysis and comments; all authors participated in the preparation of this manuscript.

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Authors and Affiliations

  1. Division of Gastroenterology/Liver Center, Department of Medicine, Beth Israel Deaconess Medical Center, Harvard Medical School, 110 Francis Street, Suite 4 A, Boston, MA, 02215, USA

    Carolina F. M. G. Pimentel, Zhenghui G. Jiang, Takeshi Otsubo, Linda Feldbrügge, Simon Robson, Nezam Afdhal & Michelle Lai

  2. Division of Pathology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, USA

    Tracy L. Challies & Imad Nasser

Authors
  1. Carolina F. M. G. Pimentel
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  2. Zhenghui G. Jiang
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  3. Takeshi Otsubo
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  4. Linda Feldbrügge
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  9. Michelle Lai
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Correspondence to Carolina F. M. G. Pimentel.

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Pimentel, C.F.M.G., Jiang, Z.G., Otsubo, T. et al. Poor Inter-test Reliability Between CK18 Kits as a Biomarker of NASH. Dig Dis Sci 61, 905–912 (2016). https://doi.org/10.1007/s10620-015-3916-1

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  • DOI: https://doi.org/10.1007/s10620-015-3916-1

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