Tumor plasticity in macrophage niches promotes dormancy, EMT and chemoresistance of non-small cell lung cancer stem cells
4 affiliations
- 1Department of Oncology and Molecular Medicine, Istituto Superiore di Sanità, Rome, Italy
- 2Department of Health Promotion, Mother and Child Care, Internal Medicine and Medical Specialties (PROMISE), University of Palermo, Palermo, Italy
- 3Villa Sofia-Cervello Hospital, Palermo, Italy
- 4Core Facilities Technical-Scientific Service, Istituto Superiore di Sanità, Rome, Italy
Abstract
Macrophages are the major immune cell type in the microenvironment of non-small cell lung cancer (NSCLC), playing a key role in tumor development, progression, and response to therapies. Elucidating the interplay between macrophages and NSCLC is crucial to fulfill the potential of macrophage-targeting strategies. We used NSCLC surgical specimens to generate multicellular spheroids enriched in cancer stem cells (CSCs) and highly responsive to microenvironmental cues. NSCLC spheroids were used to characterize state transitions induced by macrophage-tumor cell interactions in terms of proliferation, epithelial-to-mesenchymal transition (EMT), expression of stemness-related factors, self-renewal, and drug resistance. To define whether state transitions were induced by cell–cell contact or soluble factors, we compared direct co-cultures of spheroids and macrophages with transwell cultures, where cells are separated by a porous membrane. Cellular interactions were recorded by time-lapse microscopy and immunofluorescence (IF), whereas EMT, dormancy, stemness and chemoresistance were evaluated by real-time PCR, immunoblotting and functional assays. We found that co-cultures allowing cell–cell contact, but not transwell cultures, induced robust state transitions in NSCLC CSCs, promoting a state of dormancy associated with increased stemness, EMT and chemoresistance. In direct co-cultures (named NIDO, for Niche-Induced DOrmancy), macrophages and tumor cells formed nest-shaped aggregates and underwent a specular modulation of TGF-β levels, suggesting a physical transfer of this cytokine from macrophages to NSCLC cells within cellular nests. Treatment with Bafilomycin A1, an inhibitor of autophagosome-lysosome fusion and autolysosome acidification, inhibited TGF-β relocation and prevented the full spectrum of TGF-β-induced state transitions, including chemoresistance, EMT, and nest formation. By contrast, antibody-mediated TGF-β neutralization had only a partial effect, indicating secretory autophagy as a key mechanism underlying macrophage-tumor cell interplay. Altogether, these results show that lung macrophages crucially influence CSC phenotype, proliferation, and therapy resistance, pointing to a key role of local cell circuits in regulating tumor cell plasticity.
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BibTeX
@article{zeuner2026tumor,
title = {{Tumor plasticity in macrophage niches promotes dormancy, EMT and chemoresistance of non-small cell lung cancer stem cells}},
author = {Ann Zeuner and Giovanni Sette and Stefania Rossi and Elena Landolfo and Alessandro Vici and Rachele Rossi and Matilde Todaro and Alice Turdo and Vincenzo L. Lentini and Michele Signore and Alessandra Boe and Eleonora Petrucci and Maria Laura De Angelis and Federica Francescangeli},
journal = {Cell Death Discovery},
year = {2026},
month = jul,
doi = {10.1038/s41420-026-03261-1},
url = {https://doi.org/10.1038/s41420-026-03261-1},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Tumor plasticity in macrophage niches promotes dormancy, EMT and chemoresistance of non-small cell lung cancer stem cells
AU - Ann Zeuner
AU - Giovanni Sette
AU - Stefania Rossi
AU - Elena Landolfo
AU - Alessandro Vici
AU - Rachele Rossi
AU - Matilde Todaro
AU - Alice Turdo
AU - Vincenzo L. Lentini
AU - Michele Signore
AU - Alessandra Boe
AU - Eleonora Petrucci
AU - Maria Laura De Angelis
AU - Federica Francescangeli
PY - 2026
DA - 2026/07/31
JO - Cell Death Discovery
DO - 10.1038/s41420-026-03261-1
UR - https://doi.org/10.1038/s41420-026-03261-1
AB - Macrophages are the major immune cell type in the microenvironment of non-small cell lung cancer (NSCLC), playing a key role in tumor development, progression, and response to therapies. Elucidating the interplay between macrophages and NSCLC is crucial to fulfill the potential of macrophage-targeting strategies. We used NSCLC surgical specimens to generate multicellular spheroids enriched in cancer stem cells (CSCs) and highly responsive to microenvironmental cues. NSCLC spheroids were used to characterize state transitions induced by macrophage-tumor cell interactions in terms of proliferation, epithelial-to-mesenchymal transition (EMT), expression of stemness-related factors, self-renewal, and drug resistance. To define whether state transitions were induced by cell–cell contact or soluble factors, we compared direct co-cultures of spheroids and macrophages with transwell cultures, where cells are separated by a porous membrane. Cellular interactions were recorded by time-lapse microscopy and immunofluorescence (IF), whereas EMT, dormancy, stemness and chemoresistance were evaluated by real-time PCR, immunoblotting and functional assays. We found that co-cultures allowing cell–cell contact, but not transwell cultures, induced robust state transitions in NSCLC CSCs, promoting a state of dormancy associated with increased stemness, EMT and chemoresistance. In direct co-cultures (named NIDO, for Niche-Induced DOrmancy), macrophages and tumor cells formed nest-shaped aggregates and underwent a specular modulation of TGF-β levels, suggesting a physical transfer of this cytokine from macrophages to NSCLC cells within cellular nests. Treatment with Bafilomycin A1, an inhibitor of autophagosome-lysosome fusion and autolysosome acidification, inhibited TGF-β relocation and prevented the full spectrum of TGF-β-induced state transitions, including chemoresistance, EMT, and nest formation. By contrast, antibody-mediated TGF-β neutralization had only a partial effect, indicating secretory autophagy as a key mechanism underlying macrophage-tumor cell interplay. Altogether, these results show that lung macrophages crucially influence CSC phenotype, proliferation, and therapy resistance, pointing to a key role of local cell circuits in regulating tumor cell plasticity.
SN - 2058-7716
ER -