Serum-derived extracellular vesicles from breast cancer patients contribute to differential regulation of T-cell-mediated immune-escape mechanisms in breast cancer subtypes

Frontiers in Immunology

Immunology

Andrew Tutt¹  Patrycja Gazinska²  Tony Ng³  Ioannis Roxanis⁴  Fabienne Beuron⁴  Rosalind Graham⁵  Shubhankar Sinha⁵  Atousa Khiabany⁵  Thanussuyah Alaguthurai⁶  Sheeba Irshad⁷  Rafal Matkowski⁸  Revadee Liam-Or⁹  Khuloud T. Al-Jamal⁹  Fabian Flores-Borja^1,10  Birong Zhang^1,11  You Zhou^1,11  Jose Vicencio^1,12 

[1]Breast Cancer Now Research Unit, King's College London, Guy's Hospital, London, United Kingdom

[2]Breast Cancer Now Toby Robins Research Centre, Institute of Cancer Research, London, United Kingdom

[3]Breast Cancer Now Research Unit, King's College London, Guy's Hospital, London, United Kingdom

[4]Breast Cancer Now Toby Robins Research Centre, Institute of Cancer Research, London, United Kingdom

[5]Biobank Research Group, Lukasiewicz Research Network – PORT Polish Center for Technology Development, Wroclaw, Poland

[6]Breast Cancer Now Research Unit, King's College London, Guy's Hospital, London, United Kingdom

[7]Richard Dimbleby Laboratory of Cancer Research School of Cancer and Pharmaceutical Sciences, King’s College London, London, United Kingdom

[8]UCL Cancer Institute, Paul O'Gorman Building, University College London, London, United Kingdom

[9]Breast Cancer Now Toby Robins Research Centre, Institute of Cancer Research, London, United Kingdom

[10]Breast Immunology Group, School of Cancer & Pharmaceutical Sciences, King’s College London, London, United Kingdom

[11]Breast Immunology Group, School of Cancer & Pharmaceutical Sciences, King’s College London, London, United Kingdom

[12]Breast Cancer Now Research Unit, King's College London, Guy's Hospital, London, United Kingdom

[13]Breast Immunology Group, School of Cancer & Pharmaceutical Sciences, King’s College London, London, United Kingdom

[14]Breast Cancer Now Research Unit, King's College London, Guy's Hospital, London, United Kingdom

[15]Medical Oncology, Guy's & St Thomas' NHS Trust, London, United Kingdom

[16]Breast Unit, Lower Silesian Oncology, Pulmunology and Hematology Center, Wroclaw, Poland

[17]Department of Oncology, Wroclaw Medical University, Wroclaw, Poland

[18]Institute of Pharmaceutical Science, Faculty of Life Sciences & Medicine, King's College London, London, United Kingdom

[19]Richard Dimbleby Laboratory of Cancer Research School of Cancer and Pharmaceutical Sciences, King’s College London, London, United Kingdom

[20]Systems Immunity University Research Institute and Division of Infection and Immunity, School of Medicine, Cardiff University, Cardiff, United Kingdom

[21]UCL Cancer Institute, Paul O'Gorman Building, University College London, London, United Kingdom

关键词: extracellular vesicles;    breast cancer;    T cells;    immune regulation;    tumour microenvironment;   

DOI  :  10.3389/fimmu.2023.1204224

 received in 2023-04-11, accepted in 2023-06-05,  发布年份 2023

来源: Frontiers

【 摘 要 】

BackgroundIntracellular communication within the tumour is complex and extracellular vesicles (EVs) have been identified as major contributing factors for the cell-to-cell communication in the local and distant tumour environments. Here, we examine the differential effects of breast cancer (BC) subtype-specific patient serum and cell-line derived EVs in the regulation of T cell mediated immune responses.MethodsUltracentrifugation was used to isolate EVs from sera of 63BC patients, 15 healthy volunteers and 4 human breast cancer cell lines.Longitudinal blood draws for EV isolation for patients on neoadjuvant chemotherapy was also performed.Characterization of EVs was performed by Nanoparticle Tracking Analysis (NTA), transmission electron microscopy (TEM) and immunoblotting.CD63 staining was performed on a tissue microarray of 218 BC patients.In-house bioinformatics algorithms were utilized for the computation of EV associated expression scores within The Cancer Genome Atlas (TCGA) and correlated with tumour infiltrating lymphocyte (TIL) scores. In vitro stimulation of PBMCs with EVs from serum and cell-line derived EVs was performed and changes in the immune phenotypes characterized by flow cytometry.Cytokine profiles were assessed using a 105-plex immunoassay or IL10 ELISA. ResultsPatients with triple negative breast cancers (TNBCs) exhibited the lowest number of EVs in the sera; whilst the highest was detected in ER+HER2+ cancers; reflected also in the higher level of CD63+ vesicles found within the ER+HER2+ local tumour microenvironment.Transcriptomic analysis of the TCGA data identified that samples assigned with lower EV scores had significantly higher abundance of CD4+ memory activated T cells, T follicular cells and CD8 T cells, plasma, and memory B cells; whilst samples with high EV scores were more enriched for anti-inflammatory M2 macrophages and mast cells. A negative correlation between EV expression scores and stromal TIL counts was also observed.In vitro experiments confirmed that circulating EVs within breast cancer subtypes have functionally differing immunomodulatory capabilities, with EVs from patients with the most aggressive breast cancer subtype (TNBCs) demonstrating the most immune-suppressive phenotype (decreased CD3+HLA-DR+ but increased CD3+PD-L1 T cells, increased CD4+CD127-CD25hiT regulatory cells with associated increase in IL10 cytokine production).In depth assessment of the cytokine modulation triggered by the serum/cell line derived exosomes confirmed differential inflammatory cytokine profiles across differing breast cancer subtypes. Studies using the MDA-231 TNBC breast cancer cell-line derived EVs provided further support that TNBC EVs induced the most immunosuppressive response within PBMCs.DiscussionOur study supports further investigations into how tumour derived EVs are a mechanism that cancers can exploit to promote immune suppression; and breast cancer subtypes produce EVs with differing immunomodulatory capabilities.Understanding the intracellular/extracellular pathways implicated in alteration from active to suppressed immune state may provide a promising way forward for restoring immune competence in specific breast cancer patient populations.

【 授权许可】

Unknown   
Copyright © 2023 Graham, Gazinska, Zhang, Khiabany, Sinha, Alaguthurai, Flores-Borja, Vicencio, Beuron, Roxanis, Matkowski, Liam-Or, Tutt, Ng, Al-Jamal, Zhou and Irshad