Inhibition of the CCR6-CCL20 axis prevents regulatory T cell recruitment and sensitizes head and neck squamous cell carcinoma to radiation therapy

• Published in: Cancer Immunology, Immunotherapy Vol. 72; no. 5; pp. 1089 - 1102
• Main Authors: Rutihinda, Cleopatra, Haroun, Ryma, Saidi, Nour Elhouda, Ordoñez, Juan Pablo, Naasri, Sahar, Lévesque, Dominique, Boisvert, François-Michel, Fortier, Pierre-Hugues, Belzile, Mathieu, Fradet, Laurent, Hubert-Tremblay, Vincent, Turgeon, Guy Anne, Wang, Chang Shu, Delage, Patrick, Rousseau, Étienne, Paquette, Benoît, Oweida, Ayman J.
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Springer Berlin Heidelberg 01.05.2023; Springer Nature B.V
• Subjects: Cancer Research; CCL20 protein; CCR6 protein; CD8 antigen; Cell migration; Chemokine CCL20 - genetics; Chemokine CCL20 - metabolism; Chemokines; Flow cytometry; Gene expression; Head & neck cancer; Head and neck carcinoma; Head and Neck Neoplasms - metabolism; Head and Neck Neoplasms - radiotherapy; Humans; Immunology; Immunoregulation; Immunotherapy; Infiltration; Inflammation; Lymphocytes T; Medicine; Medicine & Public Health; Metastases; Oncology; Oral cavity; Phenotypes; Proteomics; Radiation; Radiation therapy; Radioresistance; Receptors, CCR6 - genetics; Receptors, CCR6 - metabolism; Squamous cell carcinoma; Squamous Cell Carcinoma of Head and Neck - metabolism; Squamous Cell Carcinoma of Head and Neck - radiotherapy; T-Lymphocytes, Regulatory; Tumor cells; Tumor Microenvironment; Tumors; CCR6-CCL20 axis; Radioresistance; Radiotherapy; Regulatory T cells; Head and neck squamous cell carcinomas
• ISSN: 0340-7004; 1432-0851
• DOI: 10.1007/s00262-022-03313-2

Background  Radioresistance of HNSCCs remains a major challenge for effective tumor control. Combined radiotherapy (RT) and immunotherapy (IT) treatment improved survival for a subset of patients with inflamed tumors or tumors susceptible to RT-induced inflammation. To overcome radioresistance and improve treatment outcomes, an understanding of factors that suppress anti-tumor immunity is necessary. In this regard, regulatory T cells (Tregs) are critical mediators of immune suppression in HNSCCs. In this study, we investigated how radiation modulates Treg infiltration in tumors through the chemokine CCL20. We hypothesized that radiation induces CCL20 secretion resulting in Treg infiltration and suppression of anti-tumor immunity. Methods  Human and mouse HNSCC cell lines with different immune phenotypes were irradiated at doses of 2 or 10 Gy. Conditioned media, RNA and protein were collected for assessment of CCL20. qPCR was used to determine CCL20 gene expression.  In vivo , MOC2 cells were implanted into the buccal cavity of mice and the effect of neutralizing CCL20 antibody was determined alone and in combination with RT. Blood samples were collected before and after RT for analysis of CCL20. Tumor samples were analyzed by flow cytometry to determine immune infiltrates, including CD8 T cells and Tregs. Mass-spectrometry was performed to analyze proteomic changes in the tumor microenvironment after anti-CCL20 treatment. Results  Cal27 and MOC2 HNSCCs had a gene signature associated with Treg infiltration, whereas SCC9 and MOC1 tumors displayed a gene signature associated with an inflamed TME. In vitro, tumor irradiation at 10 Gy significantly induced CCL20 in Cal27 and MOC2 cells relative to control. The increase in CCL20 was associated with increased Treg migration. Neutralization of CCL20 reversed radiation-induced migration of Treg cells in vitro and decreased intratumoral Tregs in vivo. Furthermore, inhibition of CCL20 resulted in a significant decrease in tumor growth compared to control in MOC2 tumors. This effect was further enhanced after combination with RT compared to either treatment alone. Conclusion  Our results suggest that radiation promotes CCL20 secretion by tumor cells which is responsible for the attraction of Tregs. Inhibition of the CCR6-CCL20 axis prevents infiltration of Tregs in tumors and suppresses tumor growth resulting in improved response to radiation.