Chromomycin A5 induces bona fide immunogenic cell death in melanoma

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 (CA5-8).MethodsICD features and DAMPs were evaluated using several in vitro techniques with metastatic melanoma cell line (B16-F10) exposed to chromomcins A5-8 such as flow cytometry, western blot, RT-PCR and luminescence. Additionally in vivo vaccination assays with CA5-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 CA5-8 and doxorubicin exhibited ICD features such as autophagy and apoptosis, externalization of calreticulin, and releasing of HMGB1. However, CA5-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 CA5 was confirmed by vaccination of C57BL/6 mice with CA5-treated cells which activated antigen-presenting cells and T lymphocytes and stimulated antitumor activity.ConclusionCA5 induces bona fide immunogenic cell death on melanoma.

Characterization of 3D NSCLC Cell Cultures with Fibroblasts or Macrophages for Tumor Microenvironment Studies and Chemotherapy Screening

“…Studies suggest that chromomycin A5 is associated with the transcription factor T-box 2 (TBX2), and could be associated with its anti-proliferative and potential anti-metastatic effects [53]. CA5 would also act as an inducer of ICD (immunogenic cell death) [54]. Chromomycins are highly cytotoxic, and can induce autophagy in melanoma cells [55].…”

Clinical application of immunogenic cell death inducers in cancer immunotherapy: turning cold tumors hot

“…An increasing number of drugs are found to perform their antitumor role through ICD. For example, chromomycin A5 (CA5) can serve as a bona fide ICD inducer, since it enhances the activation of APCs and T-cells in syngeneic mouse models vaccinated with metastatic melanoma B16-F10 cells ( Florêncio et al, 2022 ). Additionally, pemetrexed is a multi-targeting antifolate antagonist established as the first-line chemotherapeutic drug for advanced lung adenocarcinoma, and mesothelioma which increases immune-regulatory genes and induces ICD ( Mathew et al, 2018 ; Sumiyoshi et al, 2021 ).…”

Redefining bioactive small molecules from microbial metabolites as revolutionary anticancer agents

“…IRD-009 HCT-116 cells (colorectal cancer) Cell cycle modulator (G 0 /G 1 phase arrest), induction of apoptosis via DNA double strand interaction and damage [ 92 ] Prodigiosin Tripyrrole Pigment Serratia marcescens MDA-MB-231 cells and xenograft (breast cancer), MDA-MB-468 cells (breast cancer) Induction of apoptosis and downregulation of HSP90α. Suppression of migration and invasion and induction of apoptosis and via inhibition of Wnt/ β-catenin signaling [ 93 , 94 ] Pyocin S2 Bacteriocin Pseudomonas aeruginosa HepG2 cells (hepatocellular carcinoma), IM-9 cells (B-lymphoblastoid cell line) Induction of cell death [ 95 ] Pyocyanin Phenazine Pigment Pseudomonas aeruginosa MCF7 cells (breast cancer) Induction of apoptosis and necrosis [ 96 ] Rebeccamycin/NSC 655649 Indocarbozole Lechevalieria aerocoloigenes, Saccharothrix aerocolonigenes P388 cells (leukemia), L1210 cells (leukemia), B16 tumor model (melanoma), A549 xenograft (lung adenocarcinoma), Intercalates with DNA, promotes double strand breakage, inhibits topoisomerase I [ 97 , 98 ] Resistomycin Pentacyclic Polyketide Antibiotic Streptomyces aurantiacus AAA5 HepG2 cells (hepatocellular carcinoma), HeLa cells (cervical cancer), MDA-MB-231 xenograft (breast cancer), patient derived xenograft (breast cancer) Induction of cell death. Suppression of tumor progression, metastasis, and invasion by Pellino-1 inhibition and SNAIL/SLUG degradation [ 99 , 100 ] Salinomycin Carboxylic Polyether Ionophore Streptomyces albus OVCAR-8 (ovarian cancer), mammary cancer stem cells, patient -derived CLL cells Induction of apoptosis and G1 cell cycle arrest via Skp2 destabilization and Stat...…”