Performance of the PRO-C3 collagen neo-epitope biomarker in non-alcoholic fatty liver disease

Marie Boyle; Dina Tiniakos; Jorn M Schattenberg; Vlad Ratziu; Elisabetta Bugianessi; Salvatore Petta; Claudia P Oliveira; Olivier Govaere; Ramy Younes; Stuart McPherson; Pierre Bedossa; Mette J Nielsen; Morten Karsdal; Diana Leeming; Stuart Kendrick; Quentin M Anstee

doi:10.1016/j.jhepr.2019.06.004

Performance of the PRO-C3 collagen neo-epitope biomarker in non-alcoholic fatty liver disease

JHEP Rep. 2019 Jul 4;1(3):188-198. doi: 10.1016/j.jhepr.2019.06.004. eCollection 2019 Sep.

Authors

Marie Boyle^{1

2}, Dina Tiniakos^{1

3}, Jorn M Schattenberg⁴, Vlad Ratziu⁵, Elisabetta Bugianessi⁶, Salvatore Petta⁷, Claudia P Oliveira⁸, Olivier Govaere¹, Ramy Younes^{1

6}, Stuart McPherson^{1

2}, Pierre Bedossa¹, Mette J Nielsen⁹, Morten Karsdal⁹, Diana Leeming⁹, Stuart Kendrick¹⁰, Quentin M Anstee^{1

2}

Affiliations

¹ Newcastle Liver Research Group, Institute of Cellular Medicine, Faculty of Medical Sciences, Newcastle University, Newcastle upon Tyne, United Kingdom.
² Newcastle NIHR Biomedical Research Centre, Newcastle upon Tyne Hospitals NHS Trust, Newcastle upon Tyne, United Kingdom.
³ Department of Pathology, Aretaieion Hospital, National and Kapodistrian University of Athens, Athens, Greece.
⁴ I. Department of Medicine, University Medical Centre, Mainz, Germany.
⁵ Institute of Cardiometabolism and Nutrition, Pitié-Salpêtrière Hospital, Paris, France.
⁶ Division of Gastroenterology, Department of Medical Sciences, University of Torino, Torino, Italy.
⁷ Sezione di Gastroenterologia, Di.Bi.M.I.S., University of Palermo, Italy.
⁸ Department of Gastroenterology, University of Sao Paulo School of Medicine, Sao Paulo, Brazil.
⁹ Nordic Bioscience A/S, Biomarkers and Research, Herlev, Denmark.
¹⁰ GlaxoSmithKline Research & Development Ltd, Stevenage, UK.

Abstract

There is an unmet need for non-invasive biomarkers in non-alcoholic fatty liver disease (NAFLD) that can diagnose advanced disease and identify patients suitable for clinical trials. The PRO-C3 collagen neo-epitope is a putative direct marker of fibrogenesis. We assessed the performance of PRO-C3 in a large, well-characterised international NAFLD cohort and report the development and validation of 2 novel panels for the diagnosis of advanced fibrosis (F≥3) in NAFLD, including a simplified clinical score which eliminates the need for online calculators.

Methods: Plasma PRO-C3 levels were determined in a prospectively recruited international cohort of 449 patients with biopsy diagnosed NAFLD across the full disease spectrum (F0: n = 90; F1: 100; F2: 92; F3: 101; F4: 66). The cohort was divided into a discovery group (n = 151) and a validation group (n = 298). Logistic regression was performed to establish complex (FIBC3) and simplified (ABC3D) diagnostic scores that accurately identify advanced fibrosis. Performance for each was compared to established non-invasive fibrosis scoring systems.

Results: Plasma PRO-C3 levels correlated with grade of histological steatohepatitis (r_s = 0.367, p ≪0.0001) and stage of fibrosis (r_s = 0.462, p ≪0.0001), exhibiting similar performance to current fibrosis scores such as FIB4 for the detection of F≥3 fibrosis. FIBC3 exhibited substantially improved accuracy (AUROC 0.89 and 0.83 in the discovery and validation sets, respectively) and outperformed FIB4 and other similar diagnostic panels. The simplified version, ABC3D, was concurrently developed and had comparable diagnostic accuracy (AUROC 0.88 and 0.81 in the discovery and validation sets, respectively).

Conclusion: Plasma PRO-C3 levels correlate with severity of steatohepatitis and fibrosis stage. The FIBC3 panel is an accurate tool with a single threshold value that maintains both sensitivity and specificity for the identification of F≥3 fibrosis in NAFLD, eliminating indeterminate results and outperforming commonly used non-invasive tools. A greatly simplified version (ABC3D) that is readily amenable to use in the clinic has been validated and shown to perform with similar accuracy, and may prove a useful tool in routine clinical practice.

Lay summary: We performed a comprehensive, independent evaluation of a collagen biomarker (PRO-C3) to detect and quantify liver fibrosis in patients with non-alcoholic fatty liver disease (NAFLD). We report the development of 2 diagnostic panels using PRO-C3 to identify patients with advanced fibrosis, one optimal but more complex to calculate (FIBC3), the other easier to use (ABC3D) whilst still performing well.

Keywords: Biomarker; NAFLD; NASH; PRO-C3; Steatohepatitis; fibrosis.