Exosomes from metastatic cancer cells transfer amoeboid phenotype to non-metastatic cells and increase endothelial permeability: their emerging role in tumor heterogeneity

The goal of this study was to understand if exosomes derived from high-metastatic cells may influence the behavior of less aggressive cancer cells and the properties of the endothelium. We found that metastatic colon cancer cells are able to transfer their amoeboid phenotype to isogenic primary cancer cells through exosomes, and that this morphological transition is associated with the acquisition of a more aggressive behavior. Moreover, exosomes from the metastatic line (SW620Exos) exhibited higher ability to cause endothelial hyperpermeability than exosomes from the non metastatic line (SW480Exos). SWATH-based quantitative proteomic analysis highlighted that SW620Exos are significantly enriched in cytoskeletal-associated proteins including proteins activating the RhoA/ROCK pathway, known to induce amoeboid properties and destabilization of endothelial junctions. In particular, thrombin was identified as a key mediator of the effects induced by SW620Exos in target cells, in which we also found a significant increase of RhoA activity. Overall, our results demonstrate that in a heterogeneous context exosomes released by aggressive sub-clones can contribute to accelerate tumor progression by spreading malignant properties that affect both the tumor cell plasticity and the endothelial cell behavior.

[1]  Christoph Cremer,et al.  Single-Molecule Localization Microscopy allows for the analysis of cancer metastasis-specific miRNA distribution on the nanoscale , 2015, Oncotarget.

[2]  Y. Hegerfeldt,et al.  Collective cell movement in primary melanoma explants: plasticity of cell-cell interaction, beta1-integrin function, and migration strategies. , 2002, Cancer research.

[3]  Y. Ohta,et al.  FilGAP, a Rho/Rho-associated protein kinase–regulated GTPase-activating protein for Rac, controls tumor cell migration , 2012, Molecular biology of the cell.

[4]  E. Giannoni,et al.  Mesenchymal to amoeboid transition is associated with stem-like features of melanoma cells , 2014, Cell Communication and Signaling.

[5]  F. Welch,et al.  Causes and Consequences , 2017, Nature.

[6]  Fontana Simona,et al.  Contribution of proteomics to understanding the role of tumor‐derived exosomes in cancer progression: State of the art and new perspectives , 2013 .

[7]  Peter Friedl,et al.  Molecular mechanisms of cancer cell invasion and plasticity , 2006, The British journal of dermatology.

[8]  O. Fackler,et al.  Cell motility through plasma membrane blebbing , 2008, The Journal of cell biology.

[9]  G. Mize,et al.  Protease-activated receptor mediated RhoA signaling and cytoskeletal reorganization in LNCaP cells. , 2003, Biochemistry.

[10]  Ruedi Aebersold,et al.  Mass spectrometric protein maps for biomarker discovery and clinical research , 2013, Expert review of molecular diagnostics.

[11]  J. Gavard.  Endothelial permeability and VE-cadherin , 2013, Cell adhesion & migration.

[12]  E. Sahai,et al.  ERK-MAPK signaling coordinately regulates activity of Rac1 and RhoA for tumor cell motility. , 2003, Cancer cell.

[13]  Hong-Jian Zhu,et al.  Proteome profiling of exosomes derived from human primary and metastatic colorectal cancer cells reveal differential expression of key metastatic factors and signal transduction components , 2013, Proteomics.

[14]  N. Cherdyntseva,et al.  Intratumor heterogeneity: Nature and biological significance , 2013, Biochemistry (Moscow).

[15]  György Nagy,et al.  Cellular and Molecular Life Sciences REVIEW Membrane vesicles, current state-of-the-art: emerging role of extracellular vesicles , 2022 .

[16]  D. Vestweber,et al.  VE-cadherin: the major endothelial adhesion molecule controlling cellular junctions and blood vessel formation. , 2007, Arteriosclerosis, thrombosis, and vascular biology.

[17]  S. Raimondo,et al.  Exosome-mediated crosstalk between chronic myelogenous leukemia cells and human bone marrow stromal cells triggers an interleukin 8-dependent survival of leukemia cells. , 2014, Cancer letters.

[18]  J. Xiang,et al.  Epigenetic transfer of metastatic activity by uptake of highly metastatic B16 melanoma cell-released exosomes. , 2006, Experimental oncology.

[19]  Shwu-Fan Ma,et al.  Ezrin / radixin / moesin proteins differentially regulate endothelial 1 hyperpermeability after thrombin 2 , 2013 .

[20]  T. Vomastek,et al.  Cell polarity signaling in the plasticity of cancer cell invasiveness , 2016, Oncotarget.

[21]  J. Seoane,et al.  Glioblastoma Multiforme: A Look Inside Its Heterogeneous Nature , 2014, Cancers.

[22]  E. Kohn,et al.  Role of exosomes released by chronic myelogenous leukemia cells in angiogenesis , 2012, International journal of cancer.

[23]  Yaoyang Zhang,et al.  SWATH enables precise label‐free quantification on proteome scale , 2015, Proteomics.

[24]  C. Dong,et al.  RacGAP1-driven focal adhesion formation promotes melanoma transendothelial migration through mediating adherens junction disassembly. , 2015, Biochemical and biophysical research communications.

[25]  R. Muschel,et al.  Recruitment of a myeloid cell subset (CD11b/Gr1mid) via CCL2/CCR2 promotes the development of colorectal cancer liver metastasis * , 2013, Hepatology.

[26]  G. Stamp,et al.  Validation of a model of colon cancer progression , 2000, The Journal of pathology.

[27]  J. Sleeman,et al.  Do all roads lead to Rome? Routes to metastasis development , 2011, International journal of cancer.

[28]  Daehee Hwang,et al.  Colorectal cancer cell-derived microvesicles are enriched in cell cycle-related mRNAs that promote proliferation of endothelial cells , 2009, BMC Genomics.

[29]  A. Hall,et al.  Rho GTPases and the control of cell behaviour. , 2005, Biochemical Society transactions.

[30]  Y. Gho,et al.  Quantitative proteomics of extracellular vesicles derived from human primary and metastatic colorectal cancer cells , 2012, Journal of extracellular vesicles.

[31]  Peter Friedl,et al.  Compensation mechanism in tumor cell migration , 2003, The Journal of cell biology.

[32]  Valentina R Minciacchi,et al.  Large oncosomes contain distinct protein cargo and represent a separate functional class of tumor-derived extracellular vesicles , 2015, Oncotarget.

[33]  F. Miller,et al.  Cellular interactions in metastasis , 1990, Cancer and Metastasis Reviews.

[34]  A. von Kriegsheim,et al.  RCP-driven α5β1 recycling suppresses Rac and promotes RhoA activity via the RacGAP1–IQGAP1 complex , 2013, The Journal of cell biology.

[35]  Stephanie Alexander,et al.  Cancer Invasion and the Microenvironment: Plasticity and Reciprocity , 2011, Cell.

[36]  R. Shen,et al.  Exosomes mediated pentose phosphate pathway in ovarian cancer metastasis: a proteomics analysis. , 2015, International journal of clinical and experimental pathology.

[37]  Erik Sahai,et al.  Differing modes of tumour cell invasion have distinct requirements for Rho/ROCK signalling and extracellular proteolysis , 2003, Nature Cell Biology.

[38]  A. Guha,et al.  Intercellular transfer of the oncogenic receptor EGFRvIII by microvesicles derived from tumour cells , 2008, Nature Cell Biology.

[39]  P. Tassone,et al.  Involvement of multiple myeloma cell-derived exosomes in osteoclast differentiation , 2015, Oncotarget.

[40]  J. Gavard.  Endothelial permeability and VE-cadherin , 2013, Cell adhesion & migration.

[41]  Pedro Fonseca,et al.  A novel community driven software for functional enrichment analysis of extracellular vesicles data , 2017, Journal of extracellular vesicles.

[42]  J. van Rheenen,et al.  Implications of Extracellular Vesicle Transfer on Cellular Heterogeneity in Cancer: What Are the Potential Clinical Ramifications? , 2016, Cancer research.

[43]  H. Ford,et al.  Clonal cooperativity in heterogenous cancers. , 2017, Seminars in cell & developmental biology.

[44]  D. Geerts,et al.  Rho GAPs and GEFs , 2014, Cell adhesion & migration.

[45]  C. Marshall,et al.  The plasticity of cytoskeletal dynamics underlying neoplastic cell migration. , 2010, Current opinion in cell biology.

[46]  D. Tarin,et al.  Studies on relationships between metastatic and non-metastatic tumor cell populations using lineages labeled with dominant selectable genetic markers. , 1993, The International journal of developmental biology.

[47]  S. Baylin,et al.  Cancer epigenetics: tumor heterogeneity, plasticity of stem-like states, and drug resistance. , 2014, Molecular cell.

[48]  G Ciasca,et al.  Mechanical and structural comparison between primary tumor and lymph node metastasis cells in colorectal cancer. , 2015, Soft matter.

[49]  Jacco van Rheenen,et al.  Imaging hallmarks of cancer in living mice , 2014, Nature Reviews Cancer.

[50]  Gema Moreno-Bueno,et al.  Melanoma exosomes educate bone marrow progenitor cells toward a pro-metastatic phenotype through MET , 2012, Nature Medicine.

[51]  J. G. Lyons,et al.  Cellular interactions determining the production of collagenase by a rat mammary carcinoma cell line , 1989, International journal of cancer.

[52]  K. Polyak,et al.  Tumor heterogeneity: causes and consequences. , 2010, Biochimica et biophysica acta.

[53]  I. Fidler,et al.  The seed and soil hypothesis revisited—The role of tumor‐stroma interactions in metastasis to different organs , 2011, International journal of cancer.

[54]  E. Van Obberghen-Schilling,et al.  Distinct signals via Rho GTPases and Src drive shape changes by thrombin and sphingosine-1-phosphate in endothelial cells. , 2002, Journal of cell science.

[55]  E. Kohn,et al.  Angiogenesis: role of calcium-mediated signal transduction. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[56]  L. Vermeulen,et al.  Cancer heterogeneity—a multifaceted view , 2013, EMBO reports.

[57]  S. Klarenbach,et al.  Differential Actions of PAR2 and PAR1 in Stimulating Human Endothelial Cell Exocytosis and Permeability: The Role of Rho-GTPases , 2003, Circulation research.

[58]  Qindong Tan,et al.  Sevoflurane prevents lipopolysaccharide-induced barrier dysfunction in human lung microvascular endothelial cells: Rho-mediated alterations of VE-cadherin. , 2015, Biochemical and biophysical research communications.

[59]  Jacco van Rheenen,et al.  In Vivo Imaging Reveals Extracellular Vesicle-Mediated Phenocopying of Metastatic Behavior , 2015, Cell.

[60]  G. Berx,et al.  Concepts of metastasis in flux: the stromal progression model. , 2012, Seminars in cancer biology.

[61]  Gerhard Christofori,et al.  Molecular networks that regulate cancer metastasis. , 2012, Seminars in cancer biology.

[62]  Patrick Soon-Shiong,et al.  Molecular heterogeneity in breast cancer: State of the science and implications for patient care. , 2017, Seminars in cell & developmental biology.

[63]  John G. Collard,et al.  Activation of RhoA by Thrombin in Endothelial Hyperpermeability: Role of Rho Kinase and Protein Tyrosine Kinases , 2000, Circulation research.

[64]  G. Lorusso,et al.  New insights into the mechanisms of organ-specific breast cancer metastasis. , 2012, Seminars in cancer biology.

[65]  R. García-Mata,et al.  Analysis of RhoA and Rho GEF activity in whole cells and the cell nucleus , 2011, Nature Protocols.

[66]  M. D. Wetzel,et al.  Modulation of GEF‐H1 Induced Signaling by Heparanase in Brain Metastatic Melanoma Cells , 2010, Journal of cellular biochemistry.

[67]  P. Roux,et al.  Cooperative Anti-Invasive Effect of Cdc42/Rac1 Activation and ROCK Inhibition in SW620 Colorectal Cancer Cells with Elevated Blebbing Activity , 2012, PloS one.

[68]  R. Vessella,et al.  DIAPH3 governs the cellular transition to the amoeboid tumour phenotype , 2012, EMBO molecular medicine.