Monocyte Maturation Mediators Upregulate CD83, ICAM-1 and MHC Class 1 Expression on Ewing’s Sarcoma, Enhancing T Cell Cytotoxicity

Abstract

Ewing’s sarcoma (EwS) is a pediatric solid tumor entity with low somatic mutational burden and a low rate of tumor-infiltrating T cells, indicating a low extent of immunogenicity. In EwS, immunogenicity may furthermore be significantly diminished by a predominantly M2 macrophage driven pro-tumorigenic tumor microenvironment. In the past, we demonstrated that CHM1319-specific TCR-transgenic T cells are able to control EwS growth in a preclinical mouse model as well as in a patient with metastatic disease. However, new adjuvant techniques to induce long lasting and curative CHM1319-specific TCR-transgenic T cell-mediated anti-tumor responses are needed. In this work, we sought to identify a technique to improve the cytotoxic effect of CHM1319-specific TCR-transgenic T cell by altering the immunogenic cell surface marker expression on EwS cell lines using different cytokines. We demonstrate that TNF, IL-6, IL-1β and PGE2 cause pro-immunogenic CD83, MHC class I and II as well as ICAM-1 upregulation in EwS cell lines. This observation was associated with significantly improved recognition and killing of the tumor cells by EwS-specific CHM1319/HLA-A*02:01-restricted TCR-transgenic T cells. Conclusively, we demonstrate that the induction of an inflammatory signature renders EwS more susceptible to adoptive T cell therapy. TNF, which is upregulated during inflammatory processes, is of particular translational interest as its secretion may be induced in the patients e.g., by irradiation and hyperthermia in the clinical setting. In future clinical protocols, this finding may be important to identify appropriate conditioning regimens as well as point of time for adoptive T cell-based immunotherapy in EwS patients.

T Cells Directed against the Metastatic Driver Chondromodulin-1 in Ewing Sarcoma: Comparative Engineering with CRISPR/Cas9 vs. Retroviral Gene Transfer for Adoptive Transfer

“…Furthermore, CHM1 sustains the undifferentiated and invasive phenotype of EwS, which promotes lung metastasis of EwS [ 29 ]. It is highly expressed and required for metastasis [ 29 ] and serves as an EwS-specific antigen [ 18 , 22 , 30 ].…”

Current State of Immunotherapy and Mechanisms of Immune Evasion in Ewing Sarcoma and Osteosarcoma

“…In particular, treatment with GM-CSF, IL-4, TNF, IL-6, IL-1β and PGE 2 upregulated MHC-I, ICAM-1 and CD83, and improved recognition of EwS cell lines by TSAspecific TCR transgenic T cells in vitro [74]. In clinical settings, certain treatments including irradiation and hyperthermia may induce TNF secretion and MHC-I expression and may thus render EwS cells more susceptible to MHC-I-dependent immunotherapies [74,92,93]. The MHC-I presentation on tumor cells can also be induced by adenovirus-based oncolytic virotherapy.…”

“…Targeting intracellular TSAs is also problematic, mainly because of low expression of MHC-I molecules that are required for the intracellular peptide presentation to the effector CD8 + T cells [39], see also Section 2.2. However, class I expression may pertain to advanced EwS and experimental approaches have been established to induce it [72][73][74].…”

“…For example, MHC-I expression in EwS cell lines is induced by IFNγ or mediators of dendritic cell maturation, including TNF [84,90,91]. In particular, treatment with GM-CSF, IL-4, TNF, IL-6, IL-1β and PGE 2 upregulated MHC-I, ICAM-1 and CD83, and improved recognition of EwS cell lines by TSAspecific TCR transgenic T cells in vitro [74]. In clinical settings, certain treatments including irradiation and hyperthermia may induce TNF secretion and MHC-I expression and may thus render EwS cells more susceptible to MHC-I-dependent immunotherapies [74,92,93].…”