Lymphocyte exhaustion in hepatocellular carcinoma: a dynamic evolution across disease stages

BackgroundImmune checkpoint inhibitors (ICIs) have transformed cancer therapy. However, their efficacy in hepatocellular carcinoma (HCC) is limited, highlighting the need to further explore immune microenvironments and novel biomarkers. This study examined lymphocyte populations and immune checkpoin...

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Main Authors: Carla Fuster-Anglada, Josep Corominas, Aida Marsal, Neus Llarch, Gemma Iserte, Marco Sanduzzi-Zamparelli, Alejandro Forner, Joana Ferrer-Fábrega, Victor Holguin, Albert Morales, Carolina Saavedra, Maria Reig, Loreto Boix, Montserrat Marí, Alba Díaz
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-06-01
Series:Frontiers in Immunology
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Online Access:https://www.frontiersin.org/articles/10.3389/fimmu.2025.1611365/full
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author Carla Fuster-Anglada
Carla Fuster-Anglada
Carla Fuster-Anglada
Josep Corominas
Josep Corominas
Aida Marsal
Aida Marsal
Neus Llarch
Neus Llarch
Neus Llarch
Neus Llarch
Gemma Iserte
Gemma Iserte
Gemma Iserte
Marco Sanduzzi-Zamparelli
Marco Sanduzzi-Zamparelli
Marco Sanduzzi-Zamparelli
Alejandro Forner
Alejandro Forner
Alejandro Forner
Alejandro Forner
Joana Ferrer-Fábrega
Joana Ferrer-Fábrega
Joana Ferrer-Fábrega
Joana Ferrer-Fábrega
Victor Holguin
Victor Holguin
Albert Morales
Albert Morales
Albert Morales
Carolina Saavedra
Maria Reig
Maria Reig
Maria Reig
Loreto Boix
Loreto Boix
Montserrat Marí
Montserrat Marí
Montserrat Marí
Alba Díaz
Alba Díaz
Alba Díaz
Alba Díaz
author_facet Carla Fuster-Anglada
Carla Fuster-Anglada
Carla Fuster-Anglada
Josep Corominas
Josep Corominas
Aida Marsal
Aida Marsal
Neus Llarch
Neus Llarch
Neus Llarch
Neus Llarch
Gemma Iserte
Gemma Iserte
Gemma Iserte
Marco Sanduzzi-Zamparelli
Marco Sanduzzi-Zamparelli
Marco Sanduzzi-Zamparelli
Alejandro Forner
Alejandro Forner
Alejandro Forner
Alejandro Forner
Joana Ferrer-Fábrega
Joana Ferrer-Fábrega
Joana Ferrer-Fábrega
Joana Ferrer-Fábrega
Victor Holguin
Victor Holguin
Albert Morales
Albert Morales
Albert Morales
Carolina Saavedra
Maria Reig
Maria Reig
Maria Reig
Loreto Boix
Loreto Boix
Montserrat Marí
Montserrat Marí
Montserrat Marí
Alba Díaz
Alba Díaz
Alba Díaz
Alba Díaz
author_sort Carla Fuster-Anglada
collection DOAJ
description BackgroundImmune checkpoint inhibitors (ICIs) have transformed cancer therapy. However, their efficacy in hepatocellular carcinoma (HCC) is limited, highlighting the need to further explore immune microenvironments and novel biomarkers. This study examined lymphocyte populations and immune checkpoint dynamics in early, advanced, and post-progression HCC to better understand immune dynamics in HCC and to help identify predictive biomarkers and immune modulation strategies.MethodsTumoral and non-tumoral liver tissues were analyzed from HCC patients across early (n=25), advanced (n=22), and advanced-beyond-progression (n=15) stages. Lymphocyte profiling was performed using immunohistochemistry and flow cytometry, focusing on NK cells, T cells, and immune exhaustion markers. An exploratory analysis of this profile and its association with disease progression and recurrence was conducted.ResultsEarly HCC exhibited higher liver-resident NK (lrNK) cell densities in non-tumor regions, which diminished with advanced stages. Increased CD56+ cell infiltration in the tumor core was associated with recurrence. Tumor region showed elevated PD-1, NKG2A, and CD39 expression in CD4+ and CD8+ T cells, indicating progressive immune exhaustion. Advanced HCC stages demonstrated altered NK cell phenotypes, with reduced cytotoxic activation (CD16) and increased residency markers (CXCR6/CD69) in tumor-isolated lymphocytes.ConclusionsProgressive immune exhaustion and dysregulation of lrNK and T cells in HCC reflect the evolution of the immune microenvironment originating in the tumor and leaking into the non-tumoral liver, progressively diminishing the cytotoxic capacity of NK and T cells. CD56+ cell density and immune checkpoint profiles are potential biomarkers for therapeutic response and disease monitoring, underscoring the need for personalized immunotherapy strategies.
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spelling doaj-art-b198fa2c04014bcdb96bce198ddb5f962025-08-20T02:32:33ZengFrontiers Media S.A.Frontiers in Immunology1664-32242025-06-011610.3389/fimmu.2025.16113651611365Lymphocyte exhaustion in hepatocellular carcinoma: a dynamic evolution across disease stagesCarla Fuster-Anglada0Carla Fuster-Anglada1Carla Fuster-Anglada2Josep Corominas3Josep Corominas4Aida Marsal5Aida Marsal6Neus Llarch7Neus Llarch8Neus Llarch9Neus Llarch10Gemma Iserte11Gemma Iserte12Gemma Iserte13Marco Sanduzzi-Zamparelli14Marco Sanduzzi-Zamparelli15Marco Sanduzzi-Zamparelli16Alejandro Forner17Alejandro Forner18Alejandro Forner19Alejandro Forner20Joana Ferrer-Fábrega21Joana Ferrer-Fábrega22Joana Ferrer-Fábrega23Joana Ferrer-Fábrega24Victor Holguin25Victor Holguin26Albert Morales27Albert Morales28Albert Morales29Carolina Saavedra30Maria Reig31Maria Reig32Maria Reig33Loreto Boix34Loreto Boix35Montserrat Marí36Montserrat Marí37Montserrat Marí38Alba Díaz39Alba Díaz40Alba Díaz41Alba Díaz42Pathology Department, Centro de Diagnóstico Biomédico (CDB), Liver Oncology Unit, Hospital Clinic Barcelona, Barcelona, SpainBarcelona Clinic Liver Cancer (BCLC) Group, Instituto De Investigaciones Biomédicas August Pi i Sunyer (IDIBAPS), Barcelona, SpainCentro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas (CIBERehd), Madrid, SpainBarcelona Clinic Liver Cancer (BCLC) Group, Instituto De Investigaciones Biomédicas August Pi i Sunyer (IDIBAPS), Barcelona, SpainCentro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas (CIBERehd), Madrid, SpainBarcelona Clinic Liver Cancer (BCLC) Group, Instituto De Investigaciones Biomédicas August Pi i Sunyer (IDIBAPS), Barcelona, SpainCentro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas (CIBERehd), Madrid, SpainBarcelona Clinic Liver Cancer (BCLC) Group, Instituto De Investigaciones Biomédicas August Pi i Sunyer (IDIBAPS), Barcelona, SpainCentro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas (CIBERehd), Madrid, SpainLiver Oncology Unit, Liver Unit, ICMDM, Hospital Clinic Barcelona, Barcelona, SpainFacultad de Medicina, Universitat de Barcelona, Barcelona, SpainBarcelona Clinic Liver Cancer (BCLC) Group, Instituto De Investigaciones Biomédicas August Pi i Sunyer (IDIBAPS), Barcelona, SpainCentro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas (CIBERehd), Madrid, SpainLiver Oncology Unit, Liver Unit, ICMDM, Hospital Clinic Barcelona, Barcelona, SpainBarcelona Clinic Liver Cancer (BCLC) Group, Instituto De Investigaciones Biomédicas August Pi i Sunyer (IDIBAPS), Barcelona, SpainCentro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas (CIBERehd), Madrid, SpainLiver Oncology Unit, Liver Unit, ICMDM, Hospital Clinic Barcelona, Barcelona, SpainBarcelona Clinic Liver Cancer (BCLC) Group, Instituto De Investigaciones Biomédicas August Pi i Sunyer (IDIBAPS), Barcelona, SpainCentro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas (CIBERehd), Madrid, SpainLiver Oncology Unit, Liver Unit, ICMDM, Hospital Clinic Barcelona, Barcelona, SpainFacultad de Medicina, Universitat de Barcelona, Barcelona, SpainBarcelona Clinic Liver Cancer (BCLC) Group, Instituto De Investigaciones Biomédicas August Pi i Sunyer (IDIBAPS), Barcelona, SpainCentro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas (CIBERehd), Madrid, SpainFacultad de Medicina, Universitat de Barcelona, Barcelona, SpainHepatobiliopancreatic Surgery and Liver and Pancreatic Transplantation Unit, Department of Surgery, Liver Oncology Unit, Instituto Clínic de Enfermedades Digestivas y Metabólicas (ICMDM), Hospital Clinic Barcelona, Barcelona, SpainBarcelona Clinic Liver Cancer (BCLC) Group, Instituto De Investigaciones Biomédicas August Pi i Sunyer (IDIBAPS), Barcelona, SpainHepatobiliopancreatic Surgery and Liver and Pancreatic Transplantation Unit, Department of Surgery, Liver Oncology Unit, Instituto Clínic de Enfermedades Digestivas y Metabólicas (ICMDM), Hospital Clinic Barcelona, Barcelona, SpainBarcelona Clinic Liver Cancer (BCLC) Group, Instituto De Investigaciones Biomédicas August Pi i Sunyer (IDIBAPS), Barcelona, SpainCentro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas (CIBERehd), Madrid, SpainDepartment of Molecular and Cellular Biomedicine, Instituto de Investigaciones Biomédicas de Barcelona, Consejo Superior de Investigaciones Científicas (IIBB-CSIC), Institut d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), Barcelona, Catalunya, SpainMedical Statistics Core Facility, Institut d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), Hospital Clinic Barcelona, Barcelona, SpainBarcelona Clinic Liver Cancer (BCLC) Group, Instituto De Investigaciones Biomédicas August Pi i Sunyer (IDIBAPS), Barcelona, SpainCentro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas (CIBERehd), Madrid, SpainLiver Oncology Unit, Liver Unit, ICMDM, Hospital Clinic Barcelona, Barcelona, SpainBarcelona Clinic Liver Cancer (BCLC) Group, Instituto De Investigaciones Biomédicas August Pi i Sunyer (IDIBAPS), Barcelona, SpainCentro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas (CIBERehd), Madrid, SpainBarcelona Clinic Liver Cancer (BCLC) Group, Instituto De Investigaciones Biomédicas August Pi i Sunyer (IDIBAPS), Barcelona, SpainCentro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas (CIBERehd), Madrid, SpainDepartment of Molecular and Cellular Biomedicine, Instituto de Investigaciones Biomédicas de Barcelona, Consejo Superior de Investigaciones Científicas (IIBB-CSIC), Institut d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), Barcelona, Catalunya, SpainPathology Department, Centro de Diagnóstico Biomédico (CDB), Liver Oncology Unit, Hospital Clinic Barcelona, Barcelona, SpainBarcelona Clinic Liver Cancer (BCLC) Group, Instituto De Investigaciones Biomédicas August Pi i Sunyer (IDIBAPS), Barcelona, SpainCentro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas (CIBERehd), Madrid, SpainFacultad de Medicina, Universitat de Barcelona, Barcelona, SpainBackgroundImmune checkpoint inhibitors (ICIs) have transformed cancer therapy. However, their efficacy in hepatocellular carcinoma (HCC) is limited, highlighting the need to further explore immune microenvironments and novel biomarkers. This study examined lymphocyte populations and immune checkpoint dynamics in early, advanced, and post-progression HCC to better understand immune dynamics in HCC and to help identify predictive biomarkers and immune modulation strategies.MethodsTumoral and non-tumoral liver tissues were analyzed from HCC patients across early (n=25), advanced (n=22), and advanced-beyond-progression (n=15) stages. Lymphocyte profiling was performed using immunohistochemistry and flow cytometry, focusing on NK cells, T cells, and immune exhaustion markers. An exploratory analysis of this profile and its association with disease progression and recurrence was conducted.ResultsEarly HCC exhibited higher liver-resident NK (lrNK) cell densities in non-tumor regions, which diminished with advanced stages. Increased CD56+ cell infiltration in the tumor core was associated with recurrence. Tumor region showed elevated PD-1, NKG2A, and CD39 expression in CD4+ and CD8+ T cells, indicating progressive immune exhaustion. Advanced HCC stages demonstrated altered NK cell phenotypes, with reduced cytotoxic activation (CD16) and increased residency markers (CXCR6/CD69) in tumor-isolated lymphocytes.ConclusionsProgressive immune exhaustion and dysregulation of lrNK and T cells in HCC reflect the evolution of the immune microenvironment originating in the tumor and leaking into the non-tumoral liver, progressively diminishing the cytotoxic capacity of NK and T cells. CD56+ cell density and immune checkpoint profiles are potential biomarkers for therapeutic response and disease monitoring, underscoring the need for personalized immunotherapy strategies.https://www.frontiersin.org/articles/10.3389/fimmu.2025.1611365/fullhepatocellular carcinomalymphocytesNK cellsimmune exhaustionsorafenib
spellingShingle Carla Fuster-Anglada
Carla Fuster-Anglada
Carla Fuster-Anglada
Josep Corominas
Josep Corominas
Aida Marsal
Aida Marsal
Neus Llarch
Neus Llarch
Neus Llarch
Neus Llarch
Gemma Iserte
Gemma Iserte
Gemma Iserte
Marco Sanduzzi-Zamparelli
Marco Sanduzzi-Zamparelli
Marco Sanduzzi-Zamparelli
Alejandro Forner
Alejandro Forner
Alejandro Forner
Alejandro Forner
Joana Ferrer-Fábrega
Joana Ferrer-Fábrega
Joana Ferrer-Fábrega
Joana Ferrer-Fábrega
Victor Holguin
Victor Holguin
Albert Morales
Albert Morales
Albert Morales
Carolina Saavedra
Maria Reig
Maria Reig
Maria Reig
Loreto Boix
Loreto Boix
Montserrat Marí
Montserrat Marí
Montserrat Marí
Alba Díaz
Alba Díaz
Alba Díaz
Alba Díaz
Lymphocyte exhaustion in hepatocellular carcinoma: a dynamic evolution across disease stages
Frontiers in Immunology
hepatocellular carcinoma
lymphocytes
NK cells
immune exhaustion
sorafenib
title Lymphocyte exhaustion in hepatocellular carcinoma: a dynamic evolution across disease stages
title_full Lymphocyte exhaustion in hepatocellular carcinoma: a dynamic evolution across disease stages
title_fullStr Lymphocyte exhaustion in hepatocellular carcinoma: a dynamic evolution across disease stages
title_full_unstemmed Lymphocyte exhaustion in hepatocellular carcinoma: a dynamic evolution across disease stages
title_short Lymphocyte exhaustion in hepatocellular carcinoma: a dynamic evolution across disease stages
title_sort lymphocyte exhaustion in hepatocellular carcinoma a dynamic evolution across disease stages
topic hepatocellular carcinoma
lymphocytes
NK cells
immune exhaustion
sorafenib
url https://www.frontiersin.org/articles/10.3389/fimmu.2025.1611365/full
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