AUTHOR=Eslami-S Zahra , Cortés-Hernández Luis Enrique , Glogovitis Ilias , Antunes-Ferreira Mafalda , D’Ambrosi Silvia , Kurma Keerthi , Garima Françoise , Cayrefourcq Laure , Best Myron G. , Koppers-Lalic Danijela , Wurdinger Thomas , Alix-Panabières Catherine 

TITLE=In vitro cross-talk between metastasis-competent circulating tumor cells and platelets in colon cancer: a malicious association during the harsh journey in the blood

JOURNAL=Frontiers in Cell and Developmental Biology

VOLUME=11

YEAR=2023

URL=https://www.frontiersin.org/journals/cell-and-developmental-biology/articles/10.3389/fcell.2023.1209846

DOI=10.3389/fcell.2023.1209846

ISSN=2296-634X

ABSTRACT=<p><bold>Background:</bold> Platelets are active players in hemostasis, coagulation and also tumorigenesis. The cross-talk between platelets and circulating tumor cells (CTCs) may have various pro-cancer effects, including promoting tumor growth, epithelial-mesenchymal transition (EMT), metastatic cell survival, adhesion, arrest and also pre-metastatic niche and metastasis formation<bold>.</bold> Interaction with CTCs might alter the platelet transcriptome. However, as CTCs are rare events, the cross-talk between CTCs and platelets is poorly understood. Here, we used our established colon CTC lines to investigate the colon CTC-platelet cross-talk <italic>in vitro</italic> and its impact on the behavior/phenotype of both cell types.</p><p><bold>Methods:</bold> We exposed platelets isolated from healthy donors to thrombin (positive control) or to conditioned medium from three CTC lines from one patient with colon cancer and then we monitored the morphological and protein expression changes by microscopy and flow cytometry. We then analyzed the transcriptome by RNA-sequencing of platelets indirectly (presence of a Transwell insert) co-cultured with the three CTC lines. We also quantified by reverse transcription-quantitative PCR the expression of genes related to EMT and cancer development in CTCs after direct co-culture (no Transwell insert) with platelets.</p><p><bold>Results:</bold> We observed morphological and transcriptomic changes in platelets upon exposure to CTC conditioned medium and indirect co-culture (secretome). Moreover, the expression levels of genes involved in EMT (<italic>p</italic> &lt; 0.05) were decreased in CTCs co-cultured with platelets, but not of genes encoding mesenchymal markers (<italic>FN1</italic> and <italic>SNAI2</italic>). The expression levels of genes involved in cancer invasiveness (<italic>MYC, VEGFB, IL33, PTGS2</italic>, and <italic>PTGER2</italic>) were increased.</p><p><bold>Conclusion:</bold> For the first time, we studied the CTC-platelet cross-talk using our unique colon CTC lines. Incubation with CTC conditioned medium led to platelet aggregation and activation, supporting the hypothesis that their interaction may contribute to preserve CTC integrity during their journey in the bloodstream. Moreover, co-culture with platelets influenced the expression of several genes involved in invasiveness and EMT maintenance in CTCs.</p>