Molecular features of TNBC govern heterogeneity in the response to radiation and autophagy inhibition

Abstract Triple negative breast cancer (TNBC) is a heterogeneous and a highly aggressive type of breast cancer. Standard of care for TNBC patients includes surgery, radio-, chemo- and immunotherapy, depending on the stage of the disease. Immunotherapy is ineffective as monotherapy but can be enhance...

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Main Authors: Patrick Fischer, Maximilian Schmid, Anna Ohradanova-Repic, Rebecca Schneeweiss, Jana Hadatsch, Odysseus Grünert, Johannes Benedum, Anna Röhrer, Felix Staudinger, Philipp Schatzlmaier, Niccolo Bragato, Sandra Barna, Magdalena Engl, Ava Kleinwächter, Dietmar Georg, Joachim Widder, Sylvia Kerschbaum-Gruber, Dea Slade
Format: Article
Language:English
Published: Nature Publishing Group 2025-07-01
Series:Cell Death and Disease
Online Access:https://doi.org/10.1038/s41419-025-07873-w
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author Patrick Fischer
Maximilian Schmid
Anna Ohradanova-Repic
Rebecca Schneeweiss
Jana Hadatsch
Odysseus Grünert
Johannes Benedum
Anna Röhrer
Felix Staudinger
Philipp Schatzlmaier
Niccolo Bragato
Sandra Barna
Magdalena Engl
Ava Kleinwächter
Dietmar Georg
Joachim Widder
Sylvia Kerschbaum-Gruber
Dea Slade
author_facet Patrick Fischer
Maximilian Schmid
Anna Ohradanova-Repic
Rebecca Schneeweiss
Jana Hadatsch
Odysseus Grünert
Johannes Benedum
Anna Röhrer
Felix Staudinger
Philipp Schatzlmaier
Niccolo Bragato
Sandra Barna
Magdalena Engl
Ava Kleinwächter
Dietmar Georg
Joachim Widder
Sylvia Kerschbaum-Gruber
Dea Slade
author_sort Patrick Fischer
collection DOAJ
description Abstract Triple negative breast cancer (TNBC) is a heterogeneous and a highly aggressive type of breast cancer. Standard of care for TNBC patients includes surgery, radio-, chemo- and immunotherapy, depending on the stage of the disease. Immunotherapy is ineffective as monotherapy but can be enhanced with taxane chemotherapy or radiotherapy. Radiation can stimulate the immune system by activating the type I interferon (IFN-I) response through cGAS-STING signaling, which recognizes cytosolic double-stranded DNA (dsDNA). Cytosolic dsDNA can be cleared by autophagy, thereby preventing activation of cGAS-STING signaling. Autophagy inhibition was therefore proposed to potentiate the immunostimulatory effects of radiation. Here we show that different molecular features of TNBC cell lines influence the effect of X-ray and carbon ion (C-ion) irradiation and autophagy inhibition on immunogenic signaling. MDA-MB-468, with low basal autophagy and high cytosolic dsDNA, activates the IFN-I response after radiation. In contrast, MDA-MB-231, characterized by high autophagy rates and low cytosolic dsDNA, induces NF-κB signaling and CXCL10 expression upon autophagy inhibition with the VPS34 inhibitor SAR405. Autophagy inhibition in TNBC cells triggers a stronger activation of innate immune cells (monocytes, natural killer cells and dendritic cells) compared to radiation. In BRCA1-mutated MDA-MB-436 cells, C-ion irradiation was more potent compared to X-rays in inducing the NF-κB-driven immunogenic response but failed to activate immune cells. Upregulation of PD-L1 by X-rays, and especially C-ions, may contribute to reduced immune cell activation, underscoring the need for combination strategies with immune checkpoint blockade. Collectively, our study highlights the NF-κB-driven immunostimulatory effects of autophagy inhibition and the importance of understanding the molecular heterogeneity in TNBC with regard to autophagy rates, IFN-I and NF-κB signaling when designing effective treatments that target these pathways.
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spelling doaj-art-21c081c96ddf46d29b9dc8f4c5ffa4902025-08-20T03:06:10ZengNature Publishing GroupCell Death and Disease2041-48892025-07-0116111810.1038/s41419-025-07873-wMolecular features of TNBC govern heterogeneity in the response to radiation and autophagy inhibitionPatrick Fischer0Maximilian Schmid1Anna Ohradanova-Repic2Rebecca Schneeweiss3Jana Hadatsch4Odysseus Grünert5Johannes Benedum6Anna Röhrer7Felix Staudinger8Philipp Schatzlmaier9Niccolo Bragato10Sandra Barna11Magdalena Engl12Ava Kleinwächter13Dietmar Georg14Joachim Widder15Sylvia Kerschbaum-Gruber16Dea Slade17Department of Radiation Oncology, Medical University of ViennaDepartment of Radiation Oncology, Medical University of ViennaMedical University of Vienna, Center for Pathophysiology, Infectiology and Immunology, Institute for Hygiene and Applied ImmunologyMax Perutz Labs, Vienna Biocenter Campus (VBC)Max Perutz Labs, Vienna Biocenter Campus (VBC)Max Perutz Labs, Vienna Biocenter Campus (VBC)Max Perutz Labs, Vienna Biocenter Campus (VBC)Department of Radiation Oncology, Medical University of ViennaDepartment of Radiation Oncology, Medical University of ViennaMedical University of Vienna, Center for Pathophysiology, Infectiology and Immunology, Institute for Hygiene and Applied ImmunologyDepartment of Radiation Oncology, Medical University of ViennaDepartment of Radiation Oncology, Medical University of ViennaMax Perutz Labs, Vienna Biocenter Campus (VBC)Max Perutz Labs, Vienna Biocenter Campus (VBC)Department of Radiation Oncology, Medical University of ViennaDepartment of Radiation Oncology, Medical University of ViennaDepartment of Radiation Oncology, Medical University of ViennaDepartment of Radiation Oncology, Medical University of ViennaAbstract Triple negative breast cancer (TNBC) is a heterogeneous and a highly aggressive type of breast cancer. Standard of care for TNBC patients includes surgery, radio-, chemo- and immunotherapy, depending on the stage of the disease. Immunotherapy is ineffective as monotherapy but can be enhanced with taxane chemotherapy or radiotherapy. Radiation can stimulate the immune system by activating the type I interferon (IFN-I) response through cGAS-STING signaling, which recognizes cytosolic double-stranded DNA (dsDNA). Cytosolic dsDNA can be cleared by autophagy, thereby preventing activation of cGAS-STING signaling. Autophagy inhibition was therefore proposed to potentiate the immunostimulatory effects of radiation. Here we show that different molecular features of TNBC cell lines influence the effect of X-ray and carbon ion (C-ion) irradiation and autophagy inhibition on immunogenic signaling. MDA-MB-468, with low basal autophagy and high cytosolic dsDNA, activates the IFN-I response after radiation. In contrast, MDA-MB-231, characterized by high autophagy rates and low cytosolic dsDNA, induces NF-κB signaling and CXCL10 expression upon autophagy inhibition with the VPS34 inhibitor SAR405. Autophagy inhibition in TNBC cells triggers a stronger activation of innate immune cells (monocytes, natural killer cells and dendritic cells) compared to radiation. In BRCA1-mutated MDA-MB-436 cells, C-ion irradiation was more potent compared to X-rays in inducing the NF-κB-driven immunogenic response but failed to activate immune cells. Upregulation of PD-L1 by X-rays, and especially C-ions, may contribute to reduced immune cell activation, underscoring the need for combination strategies with immune checkpoint blockade. Collectively, our study highlights the NF-κB-driven immunostimulatory effects of autophagy inhibition and the importance of understanding the molecular heterogeneity in TNBC with regard to autophagy rates, IFN-I and NF-κB signaling when designing effective treatments that target these pathways.https://doi.org/10.1038/s41419-025-07873-w
spellingShingle Patrick Fischer
Maximilian Schmid
Anna Ohradanova-Repic
Rebecca Schneeweiss
Jana Hadatsch
Odysseus Grünert
Johannes Benedum
Anna Röhrer
Felix Staudinger
Philipp Schatzlmaier
Niccolo Bragato
Sandra Barna
Magdalena Engl
Ava Kleinwächter
Dietmar Georg
Joachim Widder
Sylvia Kerschbaum-Gruber
Dea Slade
Molecular features of TNBC govern heterogeneity in the response to radiation and autophagy inhibition
Cell Death and Disease
title Molecular features of TNBC govern heterogeneity in the response to radiation and autophagy inhibition
title_full Molecular features of TNBC govern heterogeneity in the response to radiation and autophagy inhibition
title_fullStr Molecular features of TNBC govern heterogeneity in the response to radiation and autophagy inhibition
title_full_unstemmed Molecular features of TNBC govern heterogeneity in the response to radiation and autophagy inhibition
title_short Molecular features of TNBC govern heterogeneity in the response to radiation and autophagy inhibition
title_sort molecular features of tnbc govern heterogeneity in the response to radiation and autophagy inhibition
url https://doi.org/10.1038/s41419-025-07873-w
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