Abstract 6814: E2F1-induced autocrine IL-6 inflammatory loop mediates cancer-immune cell crosstalk that regulates metastatic properties in melanoma
Authors: Prabir Dhar, Krishna P. Singh, Shailendra K. Gupta, Dhanya Peringot, Alf Spitschak, Brigitte M. Pützer
Journal: Cancer Research (2024)

Abstract Melanoma is a cancer that is metastatic, drug-refractory and fatal if managed late or inadequately. The transcription factor E2F1 is a key player in melanoma pathology, promoting metastatic traits like EMT, neoangiogenesis, chemoresistance and evasion of immune surveillance. Particularly, high levels of E2F1 trigger tumor progression by developing protein-protein interactions (PPI) with coregulators that enhance its potential to activate a prometastatic gene regulatory network (GRN). We aimed to investigate the extent to which E2F1-expressing melanoma cells influence the immune response. Methods: We combined transcriptomic approaches with bioinformatics and structural modeling to search for target genes involved in E2F1-activated immunomodulatory and prometastatic GRNs in aggressive melanoma. Immunoprecipitation and ChIP were performed to verify both PPI and protein-DNA interactions. Functional analyses, including qRT-PCR, immunoblotting and luciferase assays, were performed to verify expression and regulation of downstream targets. Additionally, we established a melanoma-immune cell coculture system with patient-derived CD4+ and CD8+ T cells. Cytokine arrays and ELISA were used for measurement of cytokine secretion to analyze the E2F1-induced tumor-immuno-crosstalk. Results: We identified an E2F1-dependent GRN with IL-6 as a central factor mediating EMT and immune regulation. Mechanistically, E2F1-induced activation and secretion of IL-6 leads to an autocrine inflammatory forward-feedback loop, which in turn further promotes invasiveness and EMT marker expression in melanoma cells. Interestingly, IL6 activated STAT3 associates with E2F1 in a structure-dependent manner, forming a complex that enhances IL-6 expression by binding to a composite E2F1:STAT3-responsive promoter element. Moreover, we observed that the E2F1-IL6 axis dispenses immunomodulatory effects on surrounding CD4+ and CD8+ T cells, further stimulating IL-6 release, but also leading to secretion of IL-10 into the TME. Although IL-10 has an immunosuppressive function, it also showed anti-metastatic properties in melanoma cells. Thus, in contrast to IL-6, IL-10 decreased cell motility and reduced the expression of Vim, Slug, and Snai1, while simultaneous knockdown of E2F1 increased these effects. The findings are supported by clinical data sets showing that melanoma patients who have high levels of E2F1 and IL-6 and low IL-10 exhibit poor survival. Conclusion: Our results unveil a novel gene regulatory program by which E2F1-associated IL-6 release induces the formation of an E2F1:STAT3 complex, resulting in a forward-feedback loop that promotes cancer invasion and regulates T cells response through intercellular cytokine signaling in the tumor microenvironment. Citation Format: Prabir Dhar, Krishna P. Singh, Shailendra K. Gupta, Dhanya Peringot, Alf Spitschak, Brigitte M. Pützer. E2F1-induced autocrine IL-6 inflammatory loop mediates cancer-immune cell crosstalk that regulates metastatic properties in melanoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 6814.