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Macrophage contact induces RhoA GTPase signaling to trigger tumor cell intravasation

Oncogene volume 33, pages 4203–4212 (2014)Cite this article

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Abstract

Most cancer patients die as a result of metastasis, thus it is important to understand the molecular mechanisms of dissemination, including intra- and extravasation. Although the mechanisms of extravasation have been vastly studied in vitro and in vivo, the process of intravasation is still unclear. Furthermore, how cells in the tumor microenvironment facilitate tumor cell intravasation is still unknown. Using high-resolution imaging, we found that macrophages enhance tumor cell intravasation upon physical contact. Macrophage and tumor cell contact induce RhoA activity in tumor cells, triggering the formation of actin-rich degradative protrusions called invadopodia, enabling tumor cells to degrade and break through matrix barriers during tumor cell transendothelial migration. Interestingly, we show that macrophage-induced invadopodium formation and tumor cell intravasation also occur in patient-derived tumor cells and in vivo models, revealing a conserved mechanism of tumor cell intravasation. Our results illustrate a novel heterotypic cell contact-mediated signaling role for RhoA, as well as yield mechanistic insight into the ability of cells within the tumor microenvironment to facilitate steps of the metastatic cascade.

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Acknowledgements

We thank Brian Beaty for advice regarding invadopodium experiments; David Entenberg, Vera Desmarais, Jeffrey Wyckoff and the Analytical Imaging Facility for help with microscopy; Richard Stanley, Fernando Macian, Esther Arwert, Allison Harney and Veronika Micolski for guidance and reagents; Yarong Wang for help with animal injections; and the Albert Einstein shRNA Core, the Flow Cytometry Core facility and the Histology and Comparative Pathology Core for help with experiments. We also thank Jeffrey Segall, Dianne Cox, Anne Bresnick and Aviv Bergmann for helpful discussions, and the Condeelis, Segall, Cox and Hodgson labs for advice and reagents. This work was funded by NIH CA150344 (JC), NIH CA100324 (AP), NIH GM093121 (LH and JJB-C), Post-doctoral fellowship from Susan G. Komen for the Cure© KG111405 (VPS) and NIH CA159663 (MRJ).

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Authors and Affiliations

  1. Department of Anatomy and Structural Biology, Albert Einstein College of Medicine, NY, Bronx, USA

    M Roh-Johnson, J J Bravo-Cordero, A Patsialou, V P Sharma, P Guo, L Hodgson & J Condeelis

  2. Gruss-Lipper Biophotonics Center, NY, Bronx, USA

    M Roh-Johnson, J J Bravo-Cordero, A Patsialou, V P Sharma, P Guo, L Hodgson & J Condeelis

  3. The Ben May Department for Cancer Research, University of Chicago, IL, Chicago, USA

    H Liu

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JC holds equity in and is a member of the Scientific Advisory Board of MetaStat, Inc.

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Author Contributions

MR-J and JC conceived the project idea. MR-J, JJB-C and LH performed RhoA biosensor experiments; LH cloned RhoA mutant constructs; AP maintained TN1-GFP cells, created the MDA-MB-231 dtomato cell line and performed qRT-PCR; VPS optimized protocols regarding invadopodium assays and helped with microscopy; PG and MR-J performed IMARIS 3D reconstructions; and HL created the TN1-GFP cells. MR-J performed the rest of the experiments. All authors contributed to discussions related to the manuscript. MR-J and JC wrote the manuscript.

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Roh-Johnson, M., Bravo-Cordero, J., Patsialou, A. et al. Macrophage contact induces RhoA GTPase signaling to trigger tumor cell intravasation. Oncogene 33, 4203–4212 (2014). https://doi.org/10.1038/onc.2013.377

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