title: Fibronectin on the Surface of Myeloma Cell-derived Exosomes Mediates Exosome-Cell Interactions.
abstract: Exosomes regulate cell behavior by binding to and delivering their cargo to target cells; however, the mechanisms mediating exosome-cell interactions are poorly understood. Heparan sulfates on target cell surfaces can act as receptors for exosome uptake, but the ligand for heparan sulfate on exosomes has not been identified. Using exosomes isolated from myeloma cell lines and from myeloma patients, we identify exosomal fibronectin as a key heparan sulfate-binding ligand and mediator of exosome-cell interactions. We discovered that heparan sulfate plays a dual role in exosome-cell interaction; heparan sulfate on exosomes captures fibronectin, and on target cells it acts as a receptor for fibronectin. Removal of heparan sulfate from the exosome surface releases fibronectin and dramatically inhibits exosome-target cell interaction. Antibody specific for the Hep-II heparin-binding domain of fibronectin blocks exosome interaction with tumor cells or with marrow stromal cells. Regarding exosome function, fibronectin-mediated binding of exosomes to myeloma cells activated p38 and pERK signaling and expression of downstream target genes DKK1 and MMP-9, two molecules that promote myeloma progression. Antibody against fibronectin inhibited the ability of myeloma-derived exosomes to stimulate endothelial cell invasion. Heparin or heparin mimetics including Roneparstat, a modified heparin in phase I trials in myeloma patients, significantly inhibited exosome-cell interactions. These studies provide the first evidence that fibronectin binding to heparan sulfate mediates exosome-cell interactions, revealing a fundamental mechanism important for exosome-mediated cross-talk within tumor microenvironments. Moreover, these results imply that therapeutic disruption of fibronectin-heparan sulfate interactions will negatively impact myeloma tumor growth and progression.
authors.affiliation: From the Departments of Pathology and; University of Alabama at Birmingham Comprehensive Cancer Center, University of Alabama at Birmingham, Birmingham, Alabama 35294 and. Electronic address: anuragp@uab.edu.
authors.forename: Anurag
authors.initials: A
authors.lastname: Purushothaman
authors.affiliation: From the Departments of Pathology and.
authors.forename: Shyam Kumar
authors.initials: SK
authors.lastname: Bandari
authors.affiliation: the Eshelman School of Pharmacy, University of North Carolina, Chapel Hill, North Carolina 27599.
authors.forename: Jian
authors.initials: J
authors.lastname: Liu
authors.affiliation: Surgery and.
authors.forename: James A
authors.initials: JA
authors.lastname: Mobley
authors.affiliation: From the Departments of Pathology and; University of Alabama at Birmingham Comprehensive Cancer Center, University of Alabama at Birmingham, Birmingham, Alabama 35294 and.
authors.forename: Elizabeth E
authors.initials: EE
authors.lastname: Brown
authors.affiliation: From the Departments of Pathology and; University of Alabama at Birmingham Comprehensive Cancer Center, University of Alabama at Birmingham, Birmingham, Alabama 35294 and. Electronic address: sanderson@uab.edu.
authors.forename: Ralph D
authors.initials: RD
authors.lastname: Sanderson
mesh_terms: D002450:Cell Communication; D045744:Cell Line, Tumor; D042783:Endothelial Cells / Q000166:cytology / Q000378:metabolism; D055354:Exosomes / Q000235:genetics / Q000378:metabolism*; D005353:Fibronectins / Q000235:genetics / Q000378:metabolism*; D006493:Heparin / Q000378:metabolism; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism; D020780:Matrix Metalloproteinase 9 / Q000235:genetics / Q000378:metabolism; D009101:Multiple Myeloma / Q000235:genetics / Q000378:metabolism* / Q000503:physiopathology; D015398:Signal Transduction; D048051:p38 Mitogen-Activated Protein Kinases / Q000235:genetics / Q000378:metabolism
chemical_list: C501126:DKK1 protein, human; D005353:Fibronectins; D036341:Intercellular Signaling Peptides and Proteins; D006493:Heparin; D048051:p38 Mitogen-Activated Protein Kinases; D020780:Matrix Metalloproteinase 9
keywords: exosome (vesicle); fibronectin; heparan sulfate; multiple myeloma; tumor