Published: 5 May 2022| Version 1 | DOI: 10.17632/zgp6yt5s2m.1
Francesco Missale


The pro-metastatic relevance of tumor infiltrating B-lymphocytes in laryngeal squamous cell carcinoma Abstract Objectives Laryngeal squamous cell carcinomas (LSCC) are characterized by an excellent prognosis for stage I tumors but a significant risk of locoregional and distant failure for intermediate to advanced disease. This study will investigate the clinical relevance of the tumor microenvironment in a large cohort of treatment-naïve patients affected by stage II-IV LSCC. Methods Whole slide-based digital pathology analysis was applied to measure six immune cell populations identified by immunohistochemistry (IHC) staining for CD3, CD8, CD20, CD66b, CD163 and CD38. Survival analysis was performed by Cox proportional-hazards models and unsupervised hierarchical clustering by k-means method. Double IHC staining and in-situ hybridization by RNAscope allowed further analysis of a pro-tumoral B-cells population. Results A cohort of 98 patients was enrolled and analyzed. The cluster of immune-infiltrated LSCC experienced a significantly worse disease-specific survival. As novel finding, we uncover an association between high CD20+ B-cells and a worse distant failure. The phenotypic analysis of infiltrating CD20+ B-cells showed a naïve (BCL6-CD27-Mum1-) regulatory phenotype, producing TGFß but not IL10, according to an active TGFß pathway, as proved by positivity of pSMAD2 staining. Conclusion The identification of regulatory B cells in the context of LSCC, along with the activation of the TGFß pathway, could be a further hint for new trial designs investigating the efficacy of already available molecules targeting the TGFß pathway in the context of LSCC. Keywords: laryngeal neoplasms, tumor microenvironment, B-cells, B-reg, prognosis



Brescia University, Universita degli Studi di Genova


Otorhinolaryngology, Cancer Immunology, Microenvironment, Larynx Tumor