ABSTRACT

Background and Aims Highly aggressive hepatobiliary tumours include gallbladder cancer (GBC), hepatocellular carcinoma (HCC), intrahepatic and extrahepatic cholangiocarcinoma (iCCA, eCCA) and ampulla of Vater cancer (AoV). We aimed to identify plasma biomarkers for the early diagnosis of hepatobiliary cancer by leveraging the metabolomic signatures of established clinical risk factors.

Method Based on 273,190 participants from the UK Biobank, we (1) identified metabolites associated with gallstone-related conditions (e.g. cholecystitis), primary sclerosing cholangitis (PSC) and metabolic liver diseases (e.g. cirrhosis), and (2) evaluated the relationship between the identified metabolites and the risk of GBC, HCC, iCCA, eCCA and AoV. Findings were validated in an independent group of 227,809 participants from the UK Biobank. We also derived metabolomic scores summarizing the three risk-factor signatures and evaluated their ability to stratify cancer risk.

Results We identified 27 metabolites associated with gallstone-related conditions, 11 with PSC, and 34 with metabolic liver diseases, some of which showed associations with inconsistent directions across risk factors, suggesting distinct pathogenic processes. Several metabolites were associated with cancer risk in both the discovery and validation datasets, independently of established risk factors, predominantly for HCC (16 signals) and for iCCA (4), with one for GBC and none for eCCA and AoV. Metabolomic scores clearly distinguished individuals at high risk for HCC and iCCA.

Conclusion The preselection of plasma metabolites associated with established risk factors facilitated the subsequent identification and validation of biomarkers for early cancer detection. The identified metabolites suggest specific pathogenic pathways for each type of hepatobiliary cancer. Wider replication is urgently needed to advance toward clinical implementation.

BACKGROUND AND CONTEXT Clinical risk factors for hepatobiliary cancers often progress silently, making early identification of high-risk individuals difficult and highlighting the need for biological markers detectable before clinical diagnosis.

NEW FINDINGS Risk-factor–based serum metabolomic profiling identified circulating metabolites that predict specific hepatobiliary cancers years before diagnosis, with strongest and most consistent signals for hepatocellular and intrahepatic cholangiocarcinoma.

LIMITATIONS Clinical risk factors were assumed to be frequently underdiagnosed in UK Biobank, and event numbers were relatively small for some cancers, which may have reduced power and attenuated associations for less common endpoints.

CLINICAL RESEARCH RELEVANCE This study shows that serum metabolic profiles can identify individuals at increased risk for hepatobiliary cancers long before symptoms appear, particularly for hepatocellular and intrahepatic cholangiocarcinoma. These findings support the development of precision risk-stratification strategies that may ultimately enable earlier surveillance.

BASIC RESEARCH RELEVANCE By first identifying metabolites linked to specific liver and biliary clinical conditions, the study clarifies which metabolites are indirectly associated with hepatobiliary cancers through known disease pathways. Testing these metabolites again while adjusting for diagnoses of those conditions then reveals which ones also show direct, pathway-independent associations with individual hepatobiliary cancers, providing clearer insight into cancer-specific metabolic mechanisms.

Competing Interest Statement

The authors have declared no competing interest.

Funding Statement

Funded by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) - project number 514150408 and the European Union's Horizon 2020 research and innovation program (grant 825741). We also gratefully acknowledge the data storage service SDS@hd supported by the Ministry of Science, Research, and the Arts Baden-Wuerttemberg (MWK) and the German Research Foundation (DFG) through grants INST 35/1314-1 FUGG and INST 35/1503-1 FUGG.

Author Declarations

I confirm all relevant ethical guidelines have been followed, and any necessary IRB and/or ethics committee approvals have been obtained.

Yes

The details of the IRB/oversight body that provided approval or exemption for the research described are given below:

YES. This study used human data obtained from the UK Biobank resource (Application 58030). UK Biobank provides anonymized human participant data to approved researchers following completion of required ethical and access procedures. This research involved only secondary analysis of these de‑identified human data. No direct contact with participants occurred.

I confirm that all necessary patient/participant consent has been obtained and the appropriate institutional forms have been archived, and that any patient/participant/sample identifiers included were not known to anyone (e.g., hospital staff, patients or participants themselves) outside the research group so cannot be used to identify individuals.

Yes

I understand that all clinical trials and any other prospective interventional studies must be registered with an ICMJE-approved registry, such as ClinicalTrials.gov. I confirm that any such study reported in the manuscript has been registered and the trial registration ID is provided (note: if posting a prospective study registered retrospectively, please provide a statement in the trial ID field explaining why the study was not registered in advance).

Yes

I have followed all appropriate research reporting guidelines, such as any relevant EQUATOR Network research reporting checklist(s) and other pertinent material, if applicable.

Yes

Footnotes

Grant Support: Funded by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) – project number 514150408 and the European Union’s Horizon 2020 research and innovation program (grant 825741). We also gratefully acknowledge the data storage service SDS@hd supported by the Ministry of Science, Research, and the Arts Baden-Württemberg (MWK) and the German Research Foundation (DFG) through grants INST 35/1314-1 FUGG and INST 35/1503-1 FUGG.

boekstegersfiarc.who.int, France; viallonviarc.who.int, France; marie.breeurndph.ox.ac.uk, UK; cosmin.voicanaphp.fr, France; gabriel.perlemuteraphp.fr, France; chatziioannouciarc.who.int, France; keskipiarc.who.int, France; schererimbi.uni-heidelberg.de, Germany; jenabmiarc.who.int, France; lorenzoimbi.uni-heidelberg.de, Germany

Data Availability

This research was conducted using data from UK Biobank under application number 58030. UK Biobank data are available to bona fide researchers from academic, charitable, government, and commercial institutions following project approval and completion of the required access procedures (https://www.ukbiobank.ac.uk). Access is granted for health‑related research in the public interest in accordance with UK Biobank’s data access policy.

https://www.ukbiobank.ac.uk

AbbreviationsAoVampulla of Vater cancerBMIbody mass indexeCCAextrahepatic cholangiocarcinomaGBCgallbladder canceriCCAintrahepatic cholangiocarcinomaHCChepatocellular carcinomaHDLhigh-density lipoproteinHRhazard ratioICD10the International Classification of Diseases, tenth revision, clinical modificationIDLintermediate-density lipoproteinLDLlow-density lipoproteinMASHmetabolic dysfunction-associated steatohepatitisMASLDmetabolic dysfunction-associated steatotic liver diseaseNMRnuclear magnetic resonanceOPCS-4OPCS Classification of Interventions and Procedures version 4ORodds ratioPCprincipal componentPSCprimary sclerosing cholangitisPvalprobability-valueSDstandard deviationVLDLvery low-density lipoprotein