AUTHOR=Florêncio Katharine Gurgel Dias , Edson Evelline Araújo , Fernandes Keilla Santana da Silva , Luiz João Paulo Mesquita , Pinto Francisco das Chagas Lima , Pessoa Otília Deusdênia Loiola , Cunha Fernando de Queiroz , Machado-Neto João Agostinho , Wilke Diego Veras 

TITLE=Chromomycin A5 induces bona fide immunogenic cell death in melanoma

JOURNAL=Frontiers in Immunology

VOLUME=13

YEAR=2022

URL=https://www.frontiersin.org/journals/immunology/articles/10.3389/fimmu.2022.941757

DOI=10.3389/fimmu.2022.941757

ISSN=1664-3224

ABSTRACT=<sec><title>Purpose</title><p>Some first-line cytotoxic chemotherapics, <italic>e.g.</italic> 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<sub>5-8</sub>).</p></sec><sec><title>Methods</title><p>ICD features and DAMPs were evaluated using several <italic>in vitro</italic> techniques with metastatic melanoma cell line (B16-F10) exposed to chromomcins A<sub>5-8</sub> such as flow cytometry, western blot, RT-PCR and luminescence. Additionally <italic>in vivo</italic> vaccination assays with CA<sub>5</sub>-treated cells in a syngeneic murine model (C57Bl/6) were performed to confirm ICD evaluating the immune cells activation and their antitumor activity.</p></sec><sec><title>Results</title><p>B16-F10 treated with CA<sub>5-8</sub> and doxorubicin exhibited ICD features such as autophagy and apoptosis, externalization of calreticulin, and releasing of HMGB1. However, CA<sub>5</sub>-treated cells had the best profile, also inducing ATP release, ERp57 externalization, phosphorylation of eIF2α and altering expression of transcription of genes related to autophagy, endoplasmic reticulum stress, and apoptosis. Bona fide ICD induction by CA<sub>5</sub> was confirmed by vaccination of C57BL/6 mice with CA<sub>5</sub>-treated cells which activated antigen-presenting cells and T lymphocytes and stimulated antitumor activity.</p></sec><sec><title>Conclusion</title><p>CA<sub>5</sub> induces bona fide immunogenic cell death on melanoma.</p></sec>