Hemodynamic forces tune the arrest, adhesion and extravasation of circulating tumor cells

Follain, Gautier; Osmani, Naël; Azevedo, Sofia; Allio, Guillaume; Mercier, Luc; Karreman, Matthia A.; Solecki, Gergely; Garcia‐Leon, Maria Jesus; Fekonja, Nina; Chabannes, Vincent; Dollé, Guillaume; Metivet, Thibaut; Hovsepian, François Der; Prud'Homme, Christophe; Pichot, Angélique; Paul, Nicodéme; Carapito, Raphaël; Bahram, Siamak; Ruthensteiner, Bernhard; Kemmling, André; Siemonsen, Susanne; Schneider, Tanja; Fiehler, Jens; Glatzel, Markus; Winkler, Frank; Schwab, Yannick; Pantel, Klaus; Harlepp, Sébastien; Goetz, Jacky G.

doi:10.1101/183046

Cited by 26 publications

(69 citation statements)

References 76 publications

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“…Such flow velocity was selected for two reasons. Such flow regime favors extravasation of tumor cells in zebrafish embryos and it matches the flow rate measured in capillary like vessels that we and others reported (Ivanov et al, 1985;Anton et al, 2013;Hill et al, 2015;Sugden et al, 2017;Follain et al, 2018a). Total RNAs were then extracted, quantitative RNA sequencing was carried out and flow-responsive genes were clustered and functionally annotated using Gene Ontology (GO) databases.…”

Section: Flow Upregulates Vegf Signaling Pathwaysupporting

confidence: 72%

“…The microfluidic channels were perfused with tumor cells (CTCs) that adhered to endothelial cells and further flow-stimulated for 16 hours in the presence of vehicle or sunitinib. As previously described (Follain et al, 2018a), flow stimulation favored transmigration through the endothelial cells monolayer of 95 per cent of the cancer cells. Among these, the vast majority ( 80 per cent) of the cells that had crossed the endothelial wall did so through endothelial remodeling (Fig 2C-D).…”

Section: Flow Upregulates Vegf Signaling Pathwaysupporting

confidence: 53%

“…In order to identify signaling pathways that drive endothelial remodeling in dependence of permissive flow forces, we applied RNA sequencing on cultured primary endothelial cells (HUVEC) to identify transcriptional programs activated by previously identified flow profiles. We used previously described methods based on Human Umbilical Venous Endothelial Cells (HUVEC) culture in microfluidic channels (Follain et al, 2018a). The HUVEC monolayer were cultured for 16 hours in static conditions or with medium perfusion a flow rate of 400 µm/sec ( Fig 1A).…”

Section: Flow Upregulates Vegf Signaling Pathwaymentioning

confidence: 99%

“…Thus, a comprehensive understanding of these steps is necessary in order to design relevant pharmacological therapeutic strategies. Recently, using zebrafish embryo and mouse experimental metastasis models, we have demonstrated that blood flow-induced endothelial remodeling acts as a major driving force to cell extravasation and subsequent micro-metastasis formation (Follain et al, 2018a). Specifically, we have shown that the endothelium wall actively remodels around arrested tumor cells in order to maintain vessel perfusion and thus actively promotes tumor cell extravasation.…”

Section: Introductionmentioning

confidence: 99%

“…It is currently well admitted that their development relies on biological and biophysical cues that influence their location and development (Follain et al, 2019). Indeed, adhesion molecule repertoire (Reymond et al, 2013; Osmani et al, 2019), vascular architecture (Chambers et al, 2002;Kienast et al, 2010;Headley et al, 2016) and hemodynamics (Wirtz et al, 2011;Follain et al, 2018aFollain et al, , 2020 impact the early steps of tumor cells dissemination. Several biochemical and physical parameters found in colonized organs such as growth factors, chemokines, stromal cell composition (Paget, 1889;Obenauf and Massagué, 2015;DeNardo et al, 2010;Nagarsheth et al, 2017) and matrix stiffness (Paszek et al, 2005;Levental et al, 2009;Mouw et al, 2014) will permit the growth of secondary tumors.…”

Section: Introductionmentioning

confidence: 99%

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Impairing flow-mediated endothelial remodeling reduces extravasation of tumor cells

Follain

Osmani

Mercier

et al. 2020

Preprint

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Tumor progression and metastatic dissemination are driven by cell-intrinsic and biomechanical cues that favor the growth of life-threatening secondary tumors. We recently identified prometastatic vascular regions with blood flow profiles that are permissive for the arrest of circulating tumor cells. We have further established that such flow profiles also control endothelial remodeling, which favors extravasation of arrested CTCs. Yet, how shear forces control endothelial remodeling is unknown. In the present work, we aimed at dissecting the cellular and molecular mechanisms driving blood flow-dependent endothelial remodeling. Transcriptomic analysis revealed that blood flow modulated several signaling pathways in endothelial cells. More specifically, we observed that VEGFR signaling was significantly enhanced. Using a combination of in vitro microfluidics and intravital imaging in zebrafish embryos, we now demonstrate that the early flow-driven endothelial response can be prevented with sunitinib, a pan-inhibitor of VEGFR signaling. Embryos treated with sunitinib displayed reduced endothelial remodeling and subsequent metastatic extravasation. These results confirm the importance of endothelial remodeling as a driving force of CTC extravasation and metastatic dissemination. Furthermore, the present work suggests that therapies targeting endothelial remodeling might be a relevant clinical strategy in order to impede metastatic progression.Highlights-Flow profiles that are permissive for metastasis stimulate the VEGFR pathway-Flow-dependent VEGFR signaling favors extravasation of CTCs through endothelial remodeling-Inhibition of VEGFR signaling suppresses early flow-driven endothelial response

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Section: Flow Upregulates Vegf Signaling Pathwaysupporting

confidence: 72%

Section: Flow Upregulates Vegf Signaling Pathwaysupporting

confidence: 53%

Section: Flow Upregulates Vegf Signaling Pathwaymentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Impairing flow-mediated endothelial remodeling reduces extravasation of tumor cells

Follain

Osmani

Mercier

et al. 2020

Preprint

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Monitoring circulating tumor cells in vivo by a confocal microscopy system

Tang

Cheng

et al. 2018

Cytometry Pt A

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Circulating tumor cells (CTCs) play a key role in cancer metastasis but are very difficult to detect. in vivo monitoring CTCs has been recognized as an important technique for cancer research and clinical diagnosis. Recently, a noninvasive method, in vivo flow cytometry (IVFC) has been developed to enable continuous, real-time, and long-duration detection of CTCs in animal models by detecting CTC fluorescence in blood vessels excited by lasers. In this study, we present a simple optical scheme for direct noninvasive CTC detection using confocal microscopes. We demonstrate that line scanning of confocal microscopy can provide effective and quantitative CTC detection in live mice during cancer development. Rare CTC signals can be acquired at the early stage of the tumor development after implantation of subcutaneous tumor and monitored continuously to the end. Signals from CTC clusters can also be acquired and distinguished from single CTCs. Our results suggest confocal microscopy is a simple and reliable method for biologists and doctors to use for cancer research.CIRCULATING tumor cells (CTCs) have been recognized as an important role in cancer metastasis (1). The existence of CTCs in the circulating system brings the risk that CTCs may migrate into an organ and develop a new tumor (2). Generally, in practical biological and clinical research, the in vitro assays including immunomagnetic platforms and microchips, are the most commonly used for the CTC study (3,4). The CTCs can be isolated from blood or lymph and captured by those chips. An in vivo method to capture CTCs in blood vessels was developed in 2012 by using structured medical wires (5,6). The recent observation of CTC clusters in vitro has challenged previous understandings and presented new insights on the mechanism of metastasis by CTCs (7,8). They were found with high risk of developing new tumors. The ratio of CTC clusters in total CTCs was estimated as only less than 9% in those studies but CTC clusters were believed to have higher metastatic potential than individual CTCs up to 50-fold.It is difficult to detect rare single CTCs and CTC clusters by isolation of CTCs from blood in vitro at the early stage of cancer. Moreover, it is impossible to monitor single CTCs and CTC clusters in real time by in vitro methods. Particularly, CTC clusters may be broken into single CTCs in the purification process of blood samples. In 2004, the concept of in vivo flow cytometry (IVFC) was proposed for CTC detection (9,10). The blood flow was considered as a fluidic system of cytometer and lasers were focused into the blood vessels noninvasively to excite fluorescently-labeled CTCs. The fluorescence of CTCs that passed through the laser focus could then be excited, collected and detected in vivo. Great progresses have been made by this method in cancer research (11,12). Recently, the proportion of

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Using the Zebrafish Embryo to Dissect the Early Steps of the Metastasis Cascade

Follain¹,

Osmani²,

Fuchs³

et al. 2018

Methods in Molecular Biology

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Most cancers end up with the death of patients caused by the formation of secondary tumors, called metastases. However, how these secondary tumors appear and develop is only poorly understood. A fine understanding of the multiple steps of the metastasis cascade requires in vivo models allowing high spatiotemporal analysis of the behavior of metastatic cells. Zebrafish embryos combine several advantages such as transparency, small size, stereotyped anatomy, and easy handling, making it a very powerful model for cell and cancer biology, and in vivo imaging analysis. In the following chapter, we describe a complete procedure allowing in vivo imaging methods, at high throughput and spatiotemporal resolution, to assess the behavior of circulating tumor cells (CTCs) in an experimental metastasis assay. This protocol provides access, for the first time, to the earliest steps of tumor cell seeding during metastasis formation.

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Hemodynamic forces tune the arrest, adhesion and extravasation of circulating tumor cells

Cited by 26 publications

References 76 publications

Impairing flow-mediated endothelial remodeling reduces extravasation of tumor cells

Impairing flow-mediated endothelial remodeling reduces extravasation of tumor cells

Monitoring circulating tumor cells in vivo by a confocal microscopy system

Using the Zebrafish Embryo to Dissect the Early Steps of the Metastasis Cascade

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