Extracellular vesicles promote activation of pro-inflammatory cancer-associated fibroblasts in oral cancer

Frontiers in Cell and Developmental Biology

Cell and Developmental Biology

Rebecca Towle¹  Julia Arebro²  Che-Min Lee³  Cathie Garnis⁴  Kevin L. Bennewith⁵ 

[1] Department of Integrative Oncology, British Columbia Cancer Research Center, Vancouver, BC, Canada;Department of Integrative Oncology, British Columbia Cancer Research Center, Vancouver, BC, Canada;Division of ENT Diseases, Department of Clinical Science, Intervention and Technology, Karolinska Institutet, Stockholm, Sweden;Department of ENT Diseases, Karolinska University Hospital, Stockholm, Sweden;Department of Integrative Oncology, British Columbia Cancer Research Center, Vancouver, BC, Canada;Interdisciplinary Oncology Program, University of British Columbia, Vancouver, BC, Canada;Department of Integrative Oncology, British Columbia Cancer Research Center, Vancouver, BC, Canada;Interdisciplinary Oncology Program, University of British Columbia, Vancouver, BC, Canada;Division of Otolaryngology, Department of Surgery, University of British Columbia, Vancouver, BC, Canada;Department of Integrative Oncology, British Columbia Cancer Research Center, Vancouver, BC, Canada;Interdisciplinary Oncology Program, University of British Columbia, Vancouver, BC, Canada;Pathology and Laboratory Medicine, University of British Columbia, Vancouver, BC, Canada;

关键词: cancer-associated fibroblast;    tumor microenvironment;    oral squamous cell carcinoma;    extracellular vesicles;    inflammation;    tumorigenesis;   

DOI  :  10.3389/fcell.2023.1240159

 received in 2023-06-14, accepted in 2023-08-23,  发布年份 2023

来源: Frontiers

【 摘 要 】

Introduction: Oral squamous cell carcinoma (OSCC) is the most common form of head and neck cancer and has a survival rate of ∼50% over 5 years. New treatment strategies are sorely needed to improve survival rates—and a better understanding of the mechanisms underlying tumorigenesis is needed to develop these strategies. The role of the tumor microenvironment (TME) has increasingly been identified as crucial in tumor progression and metastasis. One of the main constituents of the TME, cancer-associated fibroblasts (CAFs), plays a key role in influencing the biological behavior of tumors. Multiple mechanisms contribute to CAF activation, such as TGFβ signaling, but the role of extracellular vesicles (EVs) in CAF activation in OSCC is poorly understood. Assessing the impact of oral cancer-derived EVs on CAF activation will help to better illuminate OSCC pathophysiology and may drive development of novel treatments options.Methods: EVs were isolated from OSCC cell lines (Cal 27, SCC-9, SCC-25) using differential centrifugation. Nanoparticle tracking analysis was used for EV characterization, and Western blot to confirm the presence of EV protein markers. Oral fibroblasts were co-cultured with enriched EVs, TGFβ, or PBS over 72 h to assess activation. Flow cytometry was used to evaluate CAF markers. RNA collected from fibroblasts was extracted and the transcriptome was sequenced. Conditioned media from the co-cultures was evaluated with cytokine array profiling.Results: OSCC-derived EVs can activate oral fibroblasts into CAFs that are different from those activated by TGFβ, suggesting different mechanisms of activation and different functional properties. Gene set enrichment analysis showed several upregulated inflammatory pathways in those CAFs exposed to OSCC-derived EVs. Marker genes for inflammatory CAF subtypes were also upregulated, but not in CAFs activated by TGFβ. Finally, cytokine array analysis on secreted proteins revealed elevated levels of several pro-inflammatory cytokines from EV-activated CAFs, for instance IL-8 and CXCL5.Discussion: Our results reveal the ability of OSCC-derived EVs to activate fibroblasts into CAFs. These CAFs seem to have unique properties, differing from TGFβ-activated CAFs. Gaining an understanding of the interplay between EVs and stromal cells such as CAFs could lead to further insights into OSCC tumorigenesis and potential novel therapeutics.

【 授权许可】

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Copyright © 2023 Arebro, Towle, Lee, Bennewith and Garnis.