Tumor-derived exosomes elicit cancer-associated fibroblasts shaping inflammatory tumor microenvironment in head and neck squamous cell carcinoma

Ikko Mito; Hideyuki Takahashi; Reika Kawabata-Iwakawa; Momoka Horikawa; Shota Ida; Hiroe Tada; Toshiyuki Matsuyama; Kiyoshi Misawa; Shigeki Takeda; Kazuaki Chikamatsu

doi:10.1016/j.oraloncology.2022.106270

Tumor-derived exosomes elicit cancer-associated fibroblasts shaping inflammatory tumor microenvironment in head and neck squamous cell carcinoma

Oral Oncol. 2023 Jan:136:106270. doi: 10.1016/j.oraloncology.2022.106270. Epub 2022 Nov 30.

Affiliations

¹ Department of Otolaryngology-Head and Neck Surgery, Gunma University Graduate School of Medicine, 3-39-22, Showa-machi, Maebashi, Gunma 371-8511, Japan.
² Division of Integrated Oncology Research, Gunma University, Initiative for Advanced Research, 3-39-22, Showa-machi, Maebashi, Gunma 371-8511, Japan.
³ Faculty of Science and Technology, Division of Molecular Science, Gunma University, Kiryu, Gunma 376-8515, Japan.
⁴ Department of Otolaryngology/Head and Neck Surgery, Hamamatsu University School of Medicine, 1-20-1 Handayama, Higashi-ku, Hamamatsu, Shizuoka 431-3192, Japan.
⁵ Department of Otolaryngology-Head and Neck Surgery, Gunma University Graduate School of Medicine, 3-39-22, Showa-machi, Maebashi, Gunma 371-8511, Japan. Electronic address: tikamatu@gunma-u.ac.jp.

PMID: 36462328
DOI: 10.1016/j.oraloncology.2022.106270

Abstract

Objectives: Exosome-mediated reciprocal crosstalk between tumor and stromal cells plays a crucial role in tumor development and progression. This study investigated whether exosomes released from head and neck squamous cell carcinoma (HNSCC) tumor cells can convert normal fibroblasts into cancer-associated fibroblasts (CAF)-like cells and further analyzed the functional characterization of fibroblasts educated by tumor-derived exosomes.

Materials and methods: Exosomes secreted from HNSCC cell lines were isolated and normal fibroblasts were established from normal oropharyngeal mucosa. The effects of the exosomes on fibroblasts were examined by proliferation and migration assays, and exosome-educated fibroblasts were analyzed for the expression of eight genes (IL1B, IL6, CXCL8, TGFB1, ACTA2, FAP, CD274, and PDCD1LG2) by RT-qPCR. Moreover, T cells or CD14-positive cells were co-cultured with culture supernatants from exosome-educated fibroblasts. T-cell proliferation and macrophage polarization were examined using flow cytometry. Then, RNA sequencing (RNA-seq) of exosome-educated fibroblasts and the corresponding control fibroblasts was performed.

Results: Tumor-derived exosomes enhanced fibroblast proliferation and migration. Moreover, gene expression analysis revealed upregulation of the gene expression of proinflammatory cytokines and immunoregulatory genes, and activated fibroblast marker genes. The culture supernatants of tumor-derived exosome-educated fibroblasts suppressed T cell proliferation and the induction of protumoral macrophages compared with those of control fibroblasts. Next, comprehensive RNA-seq analysis data revealed the activation of 11 signaling pathways, including IL-6- and IL-17-related signaling.

Conclusion: These results indicate that HNSCC tumor cells induce and/or differentiate into CAFs through exosome-based cell-to-cell communication to create an inflammatory tumor microenvironment.

Keywords: Cancer-associated fibroblast; Exosome; Head and neck squamous cell carcinoma (HNSCC); Tumor microenvironment.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cancer-Associated Fibroblasts* / metabolism
Cell Line, Tumor
Cell Proliferation
Exosomes* / metabolism
Fibroblasts / metabolism
Head and Neck Neoplasms* / pathology
Humans
Squamous Cell Carcinoma of Head and Neck / pathology
Tumor Microenvironment