Determining a healthy reference range and factors potentially influencing PRO-C3 – A biomarker of liver fibrosis

Erhardtsen, Elisabeth; Rasmussen, Daniel G.K.; Frederiksen, Peder; Leeming, Diana Julie; Shevell, Diane; Gluud, Lise Lotte; Karsdal, Morten Asser; Aithal, Guruprasad P.; Schattenberg, Jörn M.

Abstract

Background & Aims
Progressive fibrosis has been identified as the major predictor of mortality in patients with non-alcoholic fatty liver disease (NAFLD). Several biomarkers are currently being evaluated for their ability to substitute the liver biopsy as the reference standard. Recent clinical studies in NAFLD/NASH patients support the utility of PRO-C3, a marker of type III collagen formation, as a marker for the degree of fibrosis, disease activity, and effect of treatment. Here we establish the healthy reference range, optimal sample handling conditions for both short- and long-term serum storage, and robustness for the PRO-C3 assay.

Methods
PRO-C3 was measured in 269 healthy volunteers and in 222 NAFLD patients. Robustness of the PRO-C3 assay was measured according to Clinical and Laboratory Standards Institute standards and included validation of interference, precision, and reagent stability, whilst sample stability was defined for storage at different temperatures and for 3 freeze-thaw cycles. Fibrosis scoring was based on histological assessments and used as a reference for the diagnostic ability of PRO-C3 to discriminate between patients with different levels of fibrosis.

Results
Robustness of the PRO-C3 analysis validated by interference, precision, and reagent stability was found to be within the predefined acceptance criteria. The healthy reference range was determined to be 6.1–14.7 ng/ml. Levels of PRO-C3 were not affected by sex, age, BMI, or ethnicity. Levels of PRO-C3 were able to identify patients with clinically significant fibrosis and advanced fibrosis (AUC = 0.83 (95% CI [0.77–0.88], p <0.0001), and AUC = 0.79 (95% CI [0.73–0.85], p <0.0001), respectively).

Conclusions
The assay proved to be robust and sample stability was found to comply with hospital sample handling requirements. PRO-C3 measured in samples from patients with NAFLD/NASH was diagnostic for significant and advanced liver fibrosis.

Citation

Erhardtsen, E., Rasmussen, D. G., Frederiksen, P., Leeming, D. J., Shevell, D., Gluud, L. L., Karsdal, M. A., Aithal, G. P., & Schattenberg, J. M. (2021). Determining a healthy reference range and factors potentially influencing PRO-C3 – A biomarker of liver fibrosis. JHEP Reports, 3(4), Article 100317. https://doi.org/10.1016/j.jhepr.2021.100317

• Journal Article Type: Article
• Acceptance Date: May 7, 2021
• Online Publication Date: Jun 4, 2021
• Publication Date: Aug 1, 2021
• Deposit Date: Jan 4, 2022
• Publicly Available Date: Jan 5, 2022
• Journal: JHEP Reports
• Electronic ISSN: 2589-5559
• Publisher: Elsevier
• Peer Reviewed: Peer Reviewed
• Volume: 3
• Issue: 4
• Article Number: 100317
• DOI: https://doi.org/10.1016/j.jhepr.2021.100317
• Keywords: Gastroenterology; Hepatology; Immunology and Allergy; Internal Medicine
• Public URL: https://nottingham-repository.worktribe.com/output/7164093
• Publisher URL: https://www.jhep-reports.eu/article/S2589-5559(21)00093-8/fulltext