Dataset Information

Secretomes from non-small cell lung cancer cells induce endothelial plasticity through a partial endothelial-to-mesenchymal transition

ABSTRACT: 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. Our study aimed to investigate the consequences of exposure to different lung tumor secretomes on EC phenotype and plasticity. Conditioned media (CM) were prepared from the tumor cell lines A549, H1755, H23, H1437 and H1975. Proliferation and migration of endothelial cells 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. A change of the endothelial phenotype was found when 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 (±-SMA and CD44) confirmed the phenotypic changes. However, the secretomes did not modify the rate of double-labelled cells (vWF+/-SMA+ or CD31+/CD44+). Proteomic analysis identified potential targets involved in the EndMT with therapeutic relevance. Taken together, these data suggest that CMs are capable of inducing partial EndMT.

INSTRUMENT(S): timsTOF Pro 2

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Endothelial Cell Of Umbilical Vein, Lung

DISEASE(S): Non-small Cell Lung Carcinoma

SUBMITTER: François Guillonneau

LAB HEAD: Nicolas Clere

PROVIDER: PXD054295 | Pride | 2025-04-24

REPOSITORIES: Pride

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Dataset's files

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			Action	DRS
	1-A549 P8_Slot1-38_2_1170.d.7z	Other
	1-A549_P8_Slot1-38_1_1169.d.7z	Other
	1-A549_P8_Slot1-38_2_1170.d.7z	Other
	2-A549 P8_Slot1-39_2_1172.d.7z	Other
	2-A549_P8_Slot1-39_1_1171.d.7z	Other

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Publications

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

Bourreau Clara C Navarro Emilie E Cotinat Marine M Krejbich Morgane M Guillonneau François F Guette Catherine C Boissard Alice A Henry Cécile C Corre Isabelle I Treps Lucas L Clere Nicolas N

Cancer medicine 20250301 5

<h4>Aim</h4>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). ...[more]

PMID: 40028673

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Project description:Hypoxia shapes the tumor microenvironment and modulates distinct cell populations activity. Low pO2 activates pathological angiogenesis in cancer- process in which endothelial cells (ECs) are the most important players. The main goal of the study was to evidence the hypoxic tumor microenvironment influence on the global gene expression pattern characteristic for ECs and evidence the distinct and specific responses displayed by the tumor-derived ECs compared to the healthy endothelium during the endothelial to mesenchymal transition (EndMT) and its regulation by miR-200-b-3p. Immortalized lines of ECs from the same patient with diagnosed breast cancer- healthy breast tissue (HBH.MEC) and primary tumor (HBCa.MEC) were used. The experiments were performed in normoxia and hypoxia for 48 hours. By wound healing test, we investigated the migration abilities of ECs. Using the established model, global gene expression analysis with NGS was carried out to detect new pathways altered in pathological ECs and to find the most changed miRNAs. The validation of NGS data from RNA and miRNA was estimated by qPCRs. Hypoxia influences ECs migration properties in wound healing assay. In hypoxia, healthy ECs migrate slower than in normoxia, as opposed to HBCa.MEC where no decreased migration ability was induced by hypoxia, due to EndMT features. NGS data identified this process to be altered in cancer ECs through extracellular matrix (ECM) organization. The deregulated genes, confirmed by qPCR, included: SPP1, ITGB6, COL4A4, ADAMST2, LAMA1, GAS6, AGTR2, PECAM1, ELN, FBLN2, COL6A3, COL9A3. NGS also identified collagens, laminin, fibronectin and integrin, as being deregulated in tumor-derived ECs. Moreover, the analysis of ten most intensively modified miRNAs when breast tumor-derived ECs were compared to healthy ECs, put a light on miR-200b-3p which is strongly up-regulated in HBCa.MECs as compared to HBH.MECs. Microenvironment influences on ECs dedifferentiation by miR-200-b-3p and ECM. The pathological ECs differed significantly, both phenotypically and functionally, from the normal corresponding tissue, thus influencing their microenvironment cross- talk. The gene expression profile confirms the EndMT phenotype of tumor-derived ECs and migratory properties acquisition. Moreover, it indicates the role of miR-200b-3p, regulating EndMT in pathological ECs, silencing several angiogenic growth factors and their receptors by directly targeting their mRNA transcripts.