Vaccination with early ferroptotic cancer cells induces efficient antitumor immunity

Journal for ImmunoTherapy of Cancer - Tập 8 Số 2 - Trang e001369 - 2020
Iuliia Efimova1,2, Elena Catanzaro3, Louis Van der Meeren4, Victoria D. Turubanova5, Hamida Hammad6,7, Tatiana A. Mishchenko5, Maria V. Vedunova5, Carmela Fimognari3, Claus Bachert8, Frauke Coppieters9, Steve Lefever9, André G. Skirtach1,4, Olga Krysko8, Dmitri V. Krysko1,2,10,5
1Cancer Research Institute Ghent, Ghent, Belgium
2Cell Death Investigation and Therapy Laboratory (CDIT), Department of Human Structure and Repair, Ghent University, Ghent, Belgium
3Department for Life Quality Studies, Alma Mater Studiorum—University of Bologna, Rimini, Italy
4NanoBioTechnology Laboratory, Department of Biotechnology, Ghent University, Ghent, Belgium
5Institute of Biology and Biomedicine, National Research Lobachevsky State University of Nizhni Novgorod, Nizhny Novgorod, Russia
6Department of Internal Medicine and Pediatrics, Ghent University, Ghent, Belgium
7Laboratory of Mucosal Immunology and Immunoregulation, VIB Center for Inflammation Research, Ghent, Belgium
8Upper Airways Research Laboratory, Department of Head and Skin, Ghent University, Ghent, Belgium
9Center for Medical Genetics Ghent (CMGG), Department of Biomolecular Medicine, Ghent University, Ghent, Belgium
10Department of Pathophysiology, Sechenov First Moscow State Medical University (Sechenov University), Moscow, Russia

Tóm tắt

Background

Immunotherapy represents the future of clinical cancer treatment. The type of cancer cell death determines the antitumor immune response and thereby contributes to the efficacy of anticancer therapy and long-term survival of patients. Induction of immunogenic apoptosis or necroptosis in cancer cells does activate antitumor immunity, but resistance to these cell death modalities is common. Therefore, it is of great importance to find other ways to kill tumor cells. Recently, ferroptosis has been identified as a novel, iron-dependent form of regulated cell death but whether ferroptotic cancer cells are immunogenic is unknown.

Methods

Ferroptotic cell death in murine fibrosarcoma MCA205 or glioma GL261 cells was induced by RAS-selective lethal 3 and ferroptosis was analyzed by flow cytometry, atomic force and confocal microscopy. ATP and high-mobility group box 1 (HMGB1) release were detected by luminescence and ELISA assays, respectively. Immunogenicity in vitro was analyzed by coculturing of ferroptotic cancer cells with bone-marrow derived dendritic cells (BMDCs) and rate of phagocytosis and activation/maturation of BMDCs (CD11c+CD86+, CD11c+CD40+, CD11c+MHCII+, IL-6, RNAseq analysis). The tumor prophylactic vaccination model in immune-competent and immune compromised (Rag-2−/−) mice was used to analyze ferroptosis immunogenicity.

Results

Ferroptosis can be induced in cancer cells by inhibition of glutathione peroxidase 4, as evidenced by confocal and atomic force microscopy and inhibitors’ analysis. We demonstrate for the first time that ferroptosis is immunogenic in vitro and in vivo. Early, but not late, ferroptotic cells promote the phenotypic maturation of BMDCs and elicit a vaccination-like effect in immune-competent mice but not in Rag-2−/− mice, suggesting that the mechanism of immunogenicity is very tightly regulated by the adaptive immune system and is time dependent. Also, ATP and HMGB1, the best-characterized damage-associated molecular patterns involved in immunogenic cell death, have proven to be passively released along the timeline of ferroptosis and act as immunogenic signal associated with the immunogenicity of early ferroptotic cancer cells.

Conclusions

These results pave the way for the development of new therapeutic strategies for cancers based on induction of ferroptosis, and thus broadens the current concept of immunogenic cell death and opens the door for the development of new strategies in cancer immunotherapy.

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