Chromomycin A(5) induces bona fide immunogenic cell death in melanoma

Author(s)

Dias Florencio, Katharine Gurgel ; Edson, Evelline Araujo ; da Silva Fernandes, Keilla Santana ; Mesquita Luiz, Joao Paulo ; Lima Pinto, Francisco das Chagas ; Loiola Pessoa, Otilia Deusdenia ; Cunha, Fernando de Queiroz ; Machado-Neto, Joao Agostinho ; Wilke, Diego Veras

• Document type: Journal article
• Source: FRONTIERS IN IMMUNOLOGY; v. 13, p. 19-pg., 2022-11-09.

Abstract

PurposeSome first-line cytotoxic chemotherapics, e.g. doxorubicin, paclitaxel and oxaliplatin, induce activation of the immune system through immunogenic cell death (ICD). Tumor cells undergoing ICD function as a vaccine, releasing damage-associated molecular patterns (DAMPs), which act as adjuvants, and neoantigens of the tumor are recognized as antigens. ICD induction is rare, however it yields better and long-lasting antitumor responses to chemotherapy. Advanced metastatic melanoma (AMM) is incurable for more than half of patients. The discovery of ICD inducers against AMM is an interesting drug discovery strategy with high translational potential. Here we evaluated ICD induction of four highly cytotoxic chromomycins A (CA(5-8)). MethodsICD features and DAMPs were evaluated using several in vitro techniques with metastatic melanoma cell line (B16-F10) exposed to chromomcins A(5-8) such as flow cytometry, western blot, RT-PCR and luminescence. Additionally in vivo vaccination assays with CA(5)-treated cells in a syngeneic murine model (C57Bl/6) were performed to confirm ICD evaluating the immune cells activation and their antitumor activity. ResultsB16-F10 treated with CA(5-8) and doxorubicin exhibited ICD features such as autophagy and apoptosis, externalization of calreticulin, and releasing of HMGB1. However, CA(5)-treated cells had the best profile, also inducing ATP release, ERp57 externalization, phosphorylation of eIF2 alpha and altering expression of transcription of genes related to autophagy, endoplasmic reticulum stress, and apoptosis. Bona fide ICD induction by CA(5) was confirmed by vaccination of C57BL/6 mice with CA(5)-treated cells which activated antigen-presenting cells and T lymphocytes and stimulated antitumor activity. ConclusionCA(5) induces bona fide immunogenic cell death on melanoma. (AU)

• FAPESP's process: 13/08216-2 - CRID - Center for Research in Inflammatory Diseases
• Grantee: Fernando de Queiroz Cunha
• Support Opportunities: Research Grants - Research, Innovation and Dissemination Centers - RIDC
• FAPESP's process: 19/23864-7 - Comprehensive analysis of genomic data for identification and validation of novel therapeutic targets involved in cellular cytoskeleton regulation in acute Leukemia
• Grantee: João Agostinho Machado Neto
• Support Opportunities: Regular Research Grants