{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,6]],"date-time":"2026-08-06T19:57:14Z","timestamp":1786046234823,"version":"3.56.0"},"reference-count":35,"publisher":"BMJ","license":[{"start":{"date-parts":[[2026,8,6]],"date-time":"2026-08-06T00:00:00Z","timestamp":1785974400000},"content-version":"unspecified","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"funder":[{"name":"German Federal Ministry of Education and Research (BMFTR","award":["01EO2106"],"award-info":[{"award-number":["01EO2106"]}]},{"DOI":"10.13039\/100008672","name":"Wilhelm Sander Foundation","doi-asserted-by":"crossref","id":[{"id":"10.13039\/100008672","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/501100001659","name":"German Research Foundation","doi-asserted-by":"crossref","id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/501100005972","name":"German Cancer Aid","doi-asserted-by":"crossref","id":[{"id":"10.13039\/501100005972","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":["bmj.com"],"crossmark-restriction":true},"short-container-title":["Gut"],"accepted":{"date-parts":[[2026,7,16]]},"abstract":"<jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>First-line immune checkpoint inhibitor (ICI) therapy has contributed to improved outcomes in hepatocellular carcinoma (HCC). However, only around 30% of patients respond, highlighting the need for biomarker-driven patient selection. Considering the lack of tissue acquirement in HCC in clinical routine, liquid biopsy holds great promise.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Objective<\/jats:title>\n                    <jats:p>We aimed at profiling immune checkpoints (IC) on circulating extracellular vesicles (EVs) as potential liquid biopsy-based biomarkers for prediction of treatment response.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Design<\/jats:title>\n                    <jats:p>Three distinct cohorts were analysed in this study: (1) HCC explorer cohort (n=40), testing the presence of membrane-bound ICs on EVs; (2) early-stage HCC cohort (n=37\u2009with paired blood and tissue), assessing the interplay between EV and tissue ICs alongside clinicopathological parameters and (3) treatment cohort (n=202\u2009with 600 sequential blood samples), comprising an ICI treated cohort with a training (n=79, n=402 sequential samples) and a validation group (n=82, n=146 sequential samples), alongside a tyrosine-kinase inhibitor (TKI) treated cohort (n=41, n=52 sequential samples), to identify predictors of immunotherapy response through IC profiling of circulating EVs via multiplex immunoassay.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>Programmed cell death protein 1 (PD-1), programmed death-ligand 1 (PD-L1) and cytotoxic T-lymphocyte associated protein 4 (CTLA-4) were enriched in EV fractions compared to EV-depleted serum. Baseline and early dynamics of EV-IC levels significantly discriminated between responders and non-responders in the ICI-treated training and validation cohorts, while simultaneously predicting progression-free survival (PFS) and overall survival (OS). These findings were not reproducible in the TKI-treated cohort. In addition, dynamic changes of EV-IC levels during therapy in patients with initial response predicted acquired therapeutic resistance, approximately 36\u201342\u2009weeks before progression was traceable on imaging.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusion<\/jats:title>\n                    <jats:p>EVs carrying PD-1, PD-L1 and CTLA-4 represent readily quantifiable, non-invasive biomarkers to predict response and survival in patients with advanced HCC undergoing ICI therapy and serve to identify biological ICI resistance much earlier than current standards with imaging.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1136\/gutjnl-2025-337617","type":"journal-article","created":{"date-parts":[[2026,8,6]],"date-time":"2026-08-06T17:20:24Z","timestamp":1786036824000},"page":"gutjnl-2025-337617","update-policy":"https:\/\/doi.org\/10.1136\/crossmarkpolicy","source":"Crossref","is-referenced-by-count":0,"title":["Circulating extracellular vesicles carrying PD-1, PD-L1 and CTLA-4 inform resistance to anti-PD-L1-based therapy in HCC"],"prefix":"10.1136","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1986-8208","authenticated-orcid":false,"given":"Jo\u00e3o","family":"Gorgulho","sequence":"first","affiliation":[{"name":"Department of Oncology, Hematology and Bone Marrow Transplantation with Section of Pneumology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany"},{"name":"Mildred Scheel Cancer Career Center, Hamburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ramsha","family":"Masood","sequence":"additional","affiliation":[{"name":"I. Department of Medicine, University Medical Centre Hamburg-Eppendorf, Hamburg, Germany"},{"name":"European Reference Network (ERN) RARE-LIVER, Hamburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gustav","family":"Buescher","sequence":"additional","affiliation":[{"name":"I. Department of Medicine, University Medical Centre Hamburg-Eppendorf, Hamburg, Germany"},{"name":"European Reference Network (ERN) RARE-LIVER, Hamburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Franziska","family":"Giehren","sequence":"additional","affiliation":[{"name":"I. Department of Medicine, University Medical Centre Hamburg-Eppendorf, Hamburg, Germany"},{"name":"European Reference Network (ERN) RARE-LIVER, Hamburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2993-3782","authenticated-orcid":false,"given":"Henrike","family":"Sali\u00e9","sequence":"additional","affiliation":[{"name":"I. Department of Medicine, University Medical Centre Hamburg-Eppendorf, Hamburg, Germany"},{"name":"European Reference Network (ERN) RARE-LIVER, Hamburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0004-2000-1753","authenticated-orcid":false,"given":"Davina","family":"David","sequence":"additional","affiliation":[{"name":"Department of Oncology, Hematology and Bone Marrow Transplantation with Section of Pneumology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany"},{"name":"I. Department of Medicine, University Medical Centre Hamburg-Eppendorf, Hamburg, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Francesca","family":"Pagani","sequence":"additional","affiliation":[{"name":"I. 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