Translating the potential of the urine steroid metabolome to stage NAFLD (TrUSt-NAFLD): study protocol for a multicentre, prospective validation study

Hamish Miller; David Harman; Guruprasad Padur Aithal; Pinelopi Manousou; Jeremy F Cobbold; Richard Parker; David Sheridan; Philip N Newsome; Fredrik Karpe; Matthew Neville; Wiebke Arlt; Alice J Sitch; Marta Korbonits; Michael Biehl; William Alazawi; Jeremy W Tomlinson

doi:10.1136/bmjopen-2023-074918

Translating the potential of the urine steroid metabolome to stage NAFLD (TrUSt-NAFLD): study protocol for a multicentre, prospective validation study

BMJ Open. 2024 Jan 18;14(1):e074918. doi: 10.1136/bmjopen-2023-074918.

Authors

Hamish Miller^{1

2}, David Harman³, Guruprasad Padur Aithal⁴, Pinelopi Manousou⁵, Jeremy F Cobbold^{6

7}, Richard Parker⁸, David Sheridan⁹, Philip N Newsome¹⁰, Fredrik Karpe¹, Matthew Neville¹, Wiebke Arlt^{11

12}, Alice J Sitch^{10

13}, Marta Korbonits¹⁴, Michael Biehl^{15

16}, William Alazawi², Jeremy W Tomlinson¹⁷

Affiliations

¹ Oxford Center for Diabetes, Endocrinology and Metabolism, University of Oxford, Oxford, UK.
² Barts Liver Centre, Queen Mary University London and Barts Health NHS Trust, London, UK.
³ Royal Berkshire Hospital NHS Foundation Trust, Reading, UK.
⁴ NIHR Nottingham Biomedical Research Centre, Nottingham University Hospitals NHS Trust and University of Nottingham, Nottingham, UK.
⁵ Department of Metabolism, Digestion and Reproduction, Imperial College London, London, UK.
⁶ Oxford Liver Unit, Oxford University Hospitals NHS Foundation Trust, Oxford, UK.
⁷ NIHR Oxford Biomedical Research Centre, Oxford University, Oxford, UK.
⁸ Leeds Liver Unit, Leeds Teaching Hospitals NHS Trust, Leeds, UK.
⁹ Institute of Translational and Stratified Medicine, University of Plymouth, Plymouth, UK.
¹⁰ National Institute for Health Research Birmingham Biomedical Research Centre, University Hospitals Birmingham NHS Foundation Trust and the University of Birmingham, Birmingham, UK.
¹¹ Institute of Metabolism and Systems Research, University of Birmingham, Birmingham, UK.
¹² Medical Research Council London Institute of Medical Sciences, Imperial College London, Hammersmith Campus, London, UK.
¹³ Institute of Applied Health Research, University of Birmingham, Birmingham, UK.
¹⁴ Centre for Endocrinology, William Harvey Research Institute, Barts and the London School of Medicine and Dentistry, Queen Mary University of London, London, UK.
¹⁵ Faculty of Science and Engineering, Bernoulli Institute for Mathematics, Computer Science and Artificial Intelligence, University of Groningen, Groningen, Netherlands.
¹⁶ SMQB, Institute of Metabolism and Systems Research, University of Birmingham, Birmingham, UK.
¹⁷ Oxford Center for Diabetes, Endocrinology and Metabolism, University of Oxford, Oxford, UK jeremy.tomlinson@ocdem.ox.ac.uk.

Abstract

Introduction: Non-alcoholic fatty liver disease (NAFLD) affects approximately one in four individuals and its prevalence continues to rise. The advanced stages of NAFLD with significant liver fibrosis are associated with adverse morbidity and mortality outcomes. Currently, liver biopsy remains the 'gold-standard' approach to stage NAFLD severity. Although generally well tolerated, liver biopsies are associated with significant complications, are resource intensive, costly, and sample only a very small area of the liver as well as requiring day case admission to a secondary care setting. As a result, there is a significant unmet need to develop non-invasive biomarkers that can accurately stage NAFLD and limit the need for liver biopsy. The aim of this study is to validate the use of the urine steroid metabolome as a strategy to stage NAFLD severity and to compare its performance against other non-invasive NAFLD biomarkers.

Methods and analysis: The TrUSt-NAFLD study is a multicentre prospective test validation study aiming to recruit 310 patients with biopsy-proven and staged NAFLD across eight centres within the UK. 150 appropriately matched control patients without liver disease will be recruited through the Oxford Biobank. Blood and urine samples, alongside clinical data, will be collected from all participants. Urine samples will be analysed by liquid chromatography-tandem mass spectroscopy to quantify a panel of predefined steroid metabolites. A machine learning-based classifier, for example, Generalized Matrix Relevance Learning Vector Quantization that was trained on retrospective samples, will be applied to the prospective steroid metabolite data to determine its ability to identify those patients with advanced, as opposed to mild-moderate, liver fibrosis as a consequence of NAFLD.

Ethics and dissemination: Research ethical approval was granted by West Midlands, Black Country Research Ethics Committee (REC reference: 21/WM/0177). A substantial amendment (TrUSt-NAFLD-SA1) was approved on 26 November 2021.

Trial registration number: ISRCTN19370855.

Keywords: DIABETES & ENDOCRINOLOGY; General endocrinology; Hepatology.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Biomarkers
Biopsy / adverse effects
Humans
Liver / pathology
Liver Cirrhosis / diagnosis
Metabolome
Multicenter Studies as Topic
Non-alcoholic Fatty Liver Disease* / epidemiology
Retrospective Studies
Steroids
Validation Studies as Topic

Substances

Biomarkers
Steroids

Associated data

ISRCTN/ISRCTN19370855

Grants and funding

WT_/Wellcome Trust/United Kingdom