title: Blocking Tumor-Educated MSC Paracrine Activity Halts Osteosarcoma Progression.
abstract: Purpose:Human osteosarcoma is a genetically heterogeneous bone malignancy with poor prognosis despite the employment of aggressive chemotherapy regimens. Because druggable driver mutations have not been established, dissecting the interactions between osteosarcoma cells and supporting stroma may provide insights into novel therapeutic targets.Experimental Design:By using a bioluminescent orthotopic xenograft mouse model of osteosarcoma, we evaluated the effect of tumor extracellular vesicle (EV)-educated mesenchymal stem cells (TEMSC) on osteosarcoma progression. Characterization and functional studies were designed to assess the mechanisms underlying MSC education. Independent series of tissue specimens were analyzed to corroborate the preclinical findings, and the composition of patient serum EVs was analyzed after isolation with size-exclusion chromatography.Results:We show that EVs secreted by highly malignant osteosarcoma cells selectively incorporate a membrane-associated form of TGFβ, which induces proinflammatory IL6 production by MSCs. TEMSCs promote tumor growth, accompanied with intratumor STAT3 activation and lung metastasis formation, which was not observed with control MSCs. Importantly, intravenous administration of the anti-IL6 receptor antibody tocilizumab abrogated the tumor-promoting effects of TEMSCs. RNA-seq analysis of human osteosarcoma tissues revealed a distinct TGFβ-induced prometastatic gene signature. Tissue microarray immunostaining indicated active STAT3 signaling in human osteosarcoma, consistent with the observations in TEMSC-treated mice. Finally, we isolated pure populations of EVs from serum and demonstrated that circulating levels of EV-associated TGFβ are increased in osteosarcoma patients.Conclusions:Collectively, our findings suggest that TEMSCs promote osteosarcoma progression and provide the basis for testing IL6- and TGFβ-blocking agents as new therapeutic options for osteosarcoma patients.Clin Cancer Res; 23(14); 3721-33. ©2017 AACR.
authors.affiliation: Department of Pathology, Cancer Center Amsterdam, VU University Medical Center, Amsterdam, the Netherlands. s.baglio@vumc.nl nicola.baldini@ior.it d.pegtel@vumc.nl.
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authors.lastname: Baglio
authors.affiliation: Department of Neurosurgery, Cancer Center Amsterdam, VU University Medical Center, Amsterdam, the Netherlands.
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authors.lastname: Lagerweij
authors.affiliation: Department of Pathology, Cancer Center Amsterdam, VU University Medical Center, Amsterdam, the Netherlands.
authors.forename: Maria
authors.initials: M
authors.lastname: Pérez-Lanzón
authors.affiliation: Department of Pathophysiology, University of Tartu, Tartu, Estonia.|Department of Traumatology and Orthopedics, University of Tartu, Tartu, Estonia.|Department of Oncology, Hue College of Medicine and Pharmacy, Hue University, Hue, Vietnam.
authors.forename: Xuan Dung
authors.initials: XD
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authors.affiliation: Laboratory for Experimental Oncology and Radiobiology, Center for Experimental Molecular Medicine (CEMM), Academic Medical Center (AMC), Amsterdam, the Netherlands.
authors.forename: Nicolas
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authors.lastname: Léveillé
authors.affiliation: Instituto de Investigação e Inovação em Saúde, Universidade do Porto (i3S) and Institute of Molecular Pathology and Immunology of the University of Porto (IPATIMUP), 4200 Porto, Portugal.
authors.forename: Sonia A
authors.initials: SA
authors.lastname: Melo
authors.affiliation: Department of Pathology, Leiden University Medical Center, Leiden, the Netherlands.
authors.forename: Anne-Marie
authors.initials: AM
authors.lastname: Cleton-Jansen
authors.affiliation: Department of Obstetrics and Gynecology, Center for Gynecological Oncology Amsterdam, VU University Medical Center, Amsterdam, the Netherlands.
authors.forename: Ekaterina S
authors.initials: ES
authors.lastname: Jordanova
authors.affiliation: Orthopaedic Pathophysiology and Regenerative Medicine Unit, Istituto Ortopedico Rizzoli, Bologna, Italy.
authors.forename: Laura
authors.initials: L
authors.lastname: Roncuzzi
authors.affiliation: Orthopaedic Pathophysiology and Regenerative Medicine Unit, Istituto Ortopedico Rizzoli, Bologna, Italy.
authors.forename: Michelina
authors.initials: M
authors.lastname: Greco
authors.affiliation: Department of Pathology, Cancer Center Amsterdam, VU University Medical Center, Amsterdam, the Netherlands.
authors.forename: Monique A J
authors.initials: MAJ
authors.lastname: van Eijndhoven
authors.affiliation: Division of Oncology, Department of Medical and Surgical Sciences for Children and Adults, University-Hospital of Modena and Reggio Emilia, Modena, Italy.
authors.forename: Giulia
authors.initials: G
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authors.affiliation: Division of Oncology, Department of Medical and Surgical Sciences for Children and Adults, University-Hospital of Modena and Reggio Emilia, Modena, Italy.
authors.forename: Massimo
authors.initials: M
authors.lastname: Dominici
authors.affiliation: Department of Pathology, Cancer Center Amsterdam, VU University Medical Center, Amsterdam, the Netherlands.|Department of Neurosciences, Biomedicine and Movement Sciences. University of Verona, Verona, Italy.
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authors.initials: R
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authors.affiliation: Department of Medical Oncology, Cancer Center Amsterdam, VU University Medical Center, Amsterdam, the Netherlands.
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authors.initials: SM
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authors.affiliation: Department of Medical Oncology, Cancer Center Amsterdam, VU University Medical Center, Amsterdam, the Netherlands.
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authors.initials: TD
authors.lastname: de Gruijl
authors.affiliation: CNR-National Research Council of Italy, Institute of Molecular Genetics, Bologna, Italy.|Laboratory of Musculoskeletal Cell Biology, Rizzoli Orthopaedic Institute, Bologna, Italy.
authors.forename: Nicoletta
authors.initials: N
authors.lastname: Zini
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authors.affiliation: CRO-Biobank, CRO Aviano National Cancer Institute, Aviano, Italy.|Clinical Cancer Pathology, CRO Aviano National Cancer Institute, Aviano, Italy.
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authors.affiliation: Orthopaedic Pathophysiology and Regenerative Medicine Unit, Istituto Ortopedico Rizzoli, Bologna, Italy. s.baglio@vumc.nl nicola.baldini@ior.it d.pegtel@vumc.nl.|Department of Biomedical and Neuromotor Sciences, University of Bologna, Bologna, Italy.
authors.forename: Nicola
authors.initials: N
authors.lastname: Baldini
authors.affiliation: Department of Pathology, Cancer Center Amsterdam, VU University Medical Center, Amsterdam, the Netherlands. s.baglio@vumc.nl nicola.baldini@ior.it d.pegtel@vumc.nl.
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mesh_terms: D000818:Animals; D061067:Antibodies, Monoclonal, Humanized / Q000008:administration & dosage; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000235:genetics; D004195:Disease Models, Animal; D000067128:Extracellular Vesicles / Q000378:metabolism; D005260:Female; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006801:Humans; D015850:Interleukin-6 / Q000235:genetics*; D008175:Lung Neoplasms / Q000188:drug therapy / Q000235:genetics* / Q000473:pathology / Q000556:secondary; D008297:Male; D059630:Mesenchymal Stem Cells / Q000378:metabolism; D051379:Mice; D012516:Osteosarcoma / Q000188:drug therapy / Q000235:genetics* / Q000473:pathology; D050796:STAT3 Transcription Factor / Q000235:genetics*; D015398:Signal Transduction / Q000235:genetics; D046888:Tissue Array Analysis; D016212:Transforming Growth Factor beta / Q000235:genetics*
chemical_list: D061067:Antibodies, Monoclonal, Humanized; C508600:IL6 protein, human; D015850:Interleukin-6; D050796:STAT3 Transcription Factor; C494086:STAT3 protein, human; D016212:Transforming Growth Factor beta; C502936:tocilizumab