Secretomes From Non‐Small Cell Lung Cancer Cells Induce Endothelial Plasticity Through a Partial Endothelial‐to‐Mesenchymal Transition

ABSTRACT Aim The tumor microenvironment (TME) of non‐small cell lung cancer (NSCLC) is highly heterogeneous and is involved in tumorigenesis and resistance to therapy. Among the cells of the TME, endothelial cells are associated with the latter processes through endothelial‐to‐mesenchymal transition...

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Main Authors: Clara Bourreau, Emilie Navarro, Marine Cotinat, Morgane Krejbich, François Guillonneau, Catherine Guette, Alice Boissard, Cécile Henry, Isabelle Corre, Lucas Treps, Nicolas Clere
Format: Article
Language:English
Published: Wiley 2025-03-01
Series:Cancer Medicine
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Online Access:https://doi.org/10.1002/cam4.70707
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author Clara Bourreau
Emilie Navarro
Marine Cotinat
Morgane Krejbich
François Guillonneau
Catherine Guette
Alice Boissard
Cécile Henry
Isabelle Corre
Lucas Treps
Nicolas Clere
author_facet Clara Bourreau
Emilie Navarro
Marine Cotinat
Morgane Krejbich
François Guillonneau
Catherine Guette
Alice Boissard
Cécile Henry
Isabelle Corre
Lucas Treps
Nicolas Clere
author_sort Clara Bourreau
collection DOAJ
description ABSTRACT Aim The tumor microenvironment (TME) of non‐small cell lung cancer (NSCLC) is highly heterogeneous and is involved in tumorigenesis and resistance to therapy. Among the cells of the TME, endothelial cells are associated with the latter processes through endothelial‐to‐mesenchymal transition (EndMT). During EndMT, endothelial cells (ECs) progressively lose their endothelial phenotype in favor of a mesenchymal phenotype, which favors the production of cancer‐associated fibroblasts (CAFs). Our study aimed to investigate the consequences of exposure to different lung tumor secretomes on EC phenotype and plasticity. Materials and Methods Conditioned media (CM) were prepared from the tumor cell lines A549, H1755, H23, H1437, and H1975. Proliferation and migration of ECs treated with these CMs were assessed by Cyquant and Incucyte technologies, respectively. The angiogenic capacity of ECs was assessed by following tubulogenesis on Matrigel. Phenotypic changes in treated ECs were detected by flow cytometry. Morphological analysis of actin fibers was performed by immunohistochemistry, while proteomic analysis by mass spectrometry was used to identify the protein content of secretomes. Results A change of the endothelial phenotype was found when human umbilical vein endothelial cells (HUVECs) were treated with different CMs. This phenotypic change was associated with a morphological change, an increase in both stress fiber expression and spontaneous migration. Furthermore, an increase in mesenchymal markers (α‐SMA and CD44) confirmed the phenotypic changes. However, the secretomes did not modify the rate of double‐labeled cells (vWF+/α‐SMA+ or CD31+/CD44+). Proteomic analysis identified potential targets involved in the EndMT with therapeutic relevance. Conclusion Taken together, these data suggest that CMs can induce partial EndMT.
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spelling doaj-art-18f05edb78c648f7b5385badf1b3ea3f2025-08-20T02:59:07ZengWileyCancer Medicine2045-76342025-03-01145n/an/a10.1002/cam4.70707Secretomes From Non‐Small Cell Lung Cancer Cells Induce Endothelial Plasticity Through a Partial Endothelial‐to‐Mesenchymal TransitionClara Bourreau0Emilie Navarro1Marine Cotinat2Morgane Krejbich3François Guillonneau4Catherine Guette5Alice Boissard6Cécile Henry7Isabelle Corre8Lucas Treps9Nicolas Clere10Univ Angers, Inserm, CNRS, MINT, SFR ICAT Angers FranceNantes Université, Université d'Angers, CHU Nantes, Inserm, CNRS, CRCI2NA Nantes FranceNantes Université, Université d'Angers, CHU Nantes, Inserm, CNRS, CRCI2NA Nantes FranceNantes Université, Université d'Angers, CHU Nantes, Inserm, CNRS, CRCI2NA Nantes FranceNantes Université, Université d'Angers, CHU Nantes, Inserm, CNRS, CRCI2NA Nantes FranceNantes Université, Université d'Angers, CHU Nantes, Inserm, CNRS, CRCI2NA Nantes FranceInstitut de Cancérologie de l'Ouest Angers FranceInstitut de Cancérologie de l'Ouest Angers FranceNantes Université, Université d'Angers, CHU Nantes, Inserm, CNRS, CRCI2NA Nantes FranceNantes Université, Université d'Angers, CHU Nantes, Inserm, CNRS, CRCI2NA Nantes FranceUniv Angers, Inserm, CNRS, MINT, SFR ICAT Angers FranceABSTRACT Aim The tumor microenvironment (TME) of non‐small cell lung cancer (NSCLC) is highly heterogeneous and is involved in tumorigenesis and resistance to therapy. Among the cells of the TME, endothelial cells are associated with the latter processes through endothelial‐to‐mesenchymal transition (EndMT). During EndMT, endothelial cells (ECs) progressively lose their endothelial phenotype in favor of a mesenchymal phenotype, which favors the production of cancer‐associated fibroblasts (CAFs). Our study aimed to investigate the consequences of exposure to different lung tumor secretomes on EC phenotype and plasticity. Materials and Methods Conditioned media (CM) were prepared from the tumor cell lines A549, H1755, H23, H1437, and H1975. Proliferation and migration of ECs treated with these CMs were assessed by Cyquant and Incucyte technologies, respectively. The angiogenic capacity of ECs was assessed by following tubulogenesis on Matrigel. Phenotypic changes in treated ECs were detected by flow cytometry. Morphological analysis of actin fibers was performed by immunohistochemistry, while proteomic analysis by mass spectrometry was used to identify the protein content of secretomes. Results A change of the endothelial phenotype was found when human umbilical vein endothelial cells (HUVECs) were treated with different CMs. This phenotypic change was associated with a morphological change, an increase in both stress fiber expression and spontaneous migration. Furthermore, an increase in mesenchymal markers (α‐SMA and CD44) confirmed the phenotypic changes. However, the secretomes did not modify the rate of double‐labeled cells (vWF+/α‐SMA+ or CD31+/CD44+). Proteomic analysis identified potential targets involved in the EndMT with therapeutic relevance. Conclusion Taken together, these data suggest that CMs can induce partial EndMT.https://doi.org/10.1002/cam4.70707EndMTendothelial cell plasticityendothelial‐to‐mesenchymal transitionnon‐small cell lung cancersecretome
spellingShingle Clara Bourreau
Emilie Navarro
Marine Cotinat
Morgane Krejbich
François Guillonneau
Catherine Guette
Alice Boissard
Cécile Henry
Isabelle Corre
Lucas Treps
Nicolas Clere
Secretomes From Non‐Small Cell Lung Cancer Cells Induce Endothelial Plasticity Through a Partial Endothelial‐to‐Mesenchymal Transition
Cancer Medicine
EndMT
endothelial cell plasticity
endothelial‐to‐mesenchymal transition
non‐small cell lung cancer
secretome
title Secretomes From Non‐Small Cell Lung Cancer Cells Induce Endothelial Plasticity Through a Partial Endothelial‐to‐Mesenchymal Transition
title_full Secretomes From Non‐Small Cell Lung Cancer Cells Induce Endothelial Plasticity Through a Partial Endothelial‐to‐Mesenchymal Transition
title_fullStr Secretomes From Non‐Small Cell Lung Cancer Cells Induce Endothelial Plasticity Through a Partial Endothelial‐to‐Mesenchymal Transition
title_full_unstemmed Secretomes From Non‐Small Cell Lung Cancer Cells Induce Endothelial Plasticity Through a Partial Endothelial‐to‐Mesenchymal Transition
title_short Secretomes From Non‐Small Cell Lung Cancer Cells Induce Endothelial Plasticity Through a Partial Endothelial‐to‐Mesenchymal Transition
title_sort secretomes from non small cell lung cancer cells induce endothelial plasticity through a partial endothelial to mesenchymal transition
topic EndMT
endothelial cell plasticity
endothelial‐to‐mesenchymal transition
non‐small cell lung cancer
secretome
url https://doi.org/10.1002/cam4.70707
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