Abstract
Purpose
Better understanding of the molecular mechanisms of metastasis—the major cause of death in osteosarcoma (OS)—is a key for the development of more effective metastasis-suppressive therapy. Here, we investigated the biological relevance of the CXCL12/CXCR4 axis in OS.
Methods
We interfered with CXCL12/CXCR4 signaling in CXCR4-expressing human 143-B OS cells through stable expression of CXCL12, of its competitive antagonist P2G, or of CXCL12-KDEL, designed to retain CXCR4 within the cell. Intratibial OS xenograft mouse model metastasizing to the lung was used to assess tumorigenic and metastatic potential of the manipulated cell lines.
Results
Constitutive expression of native CXCL12 promoted lung metastasis without affecting tumor growth. Stable expression of P2G or CXCL12-KDEL significantly accelerated tumor growth but diminished lung metastasis. Tumors grown from P2G- or CXCL12-KDEL-expressing cells contained higher levels of CXCR4-encoding mRNA going along with a higher percentage of CXCR4-expressing tumor cells. Lung metastases of all groups were predominantly enriched with CXCR4-expressing tumor cells.
Conclusion
Higher abundance of CXCR4 possibly contributed to increased local retention of tumor cells by bone marrow-derived CXCL12, reflected in the increased primary tumor growth and decreased number of lung metastases in P2G and CXCL12-KDEL groups. Higher percentage of CXCR4-expressing lung metastatic cells compared to the corresponding primary tumors point to important functions of the CXCL12/CXCR4 axis in late steps of metastasis. In conclusion, based on the here reported results, local treatment of lung metastases with novel CXCR4-targeting therapeutics might be considered and favored over anti-CXCR4 systemic therapy.
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Acknowledgments
We thank Christopher Bühler for the excellent technical assistance and Dr. Malgorzata Kisielow (ETH Zurich Flow Cytometry Facility, Zurich, Switzerland) and Dr. David Jarrossay (Università Della Svizzera Italiana, Bellinzona, Switzerland) for their help with flow cytometry analysis. We thank Prof. Kitamura and Dr. Ueda for providing us with the CXCL12-KDEL construct.
Authors’ roles
ON, MJEA, MT, and AG designed the study; ON and PB conducted the study; ON, MJEA, AK, BR, and SB were involved in the animal work; MT contributed the material; ON, MJEA, MT, and WB analyzed and interpreted the data; ON, MJEA, PB, WB, MT, and BF prepared the manuscript.
Financial support
Our work is supported by the University of Zurich, the Schweizerischer Verein Balgrist (Zurich, Switzerland), the Walter L. & Johanna Wolf Foundation (Zurich, Switzerland), the Highly Specialized Medicine for Musculoskeletal Oncology program of the Canton of Zurich, the Zurcher Krebsliga (Zurich, Switzerland), the “Kind und Krebs” fund (Zollikerberg, Switzerland), and the Swiss National Science Foundation SNF Nr. 310030_149649.
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Research involving animals: All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. Research Involving humans participants: This article does not contain any studies with human participants performed by any of the authors.
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Neklyudova, O., Arlt, M.J.E., Brennecke, P. et al. Altered CXCL12 expression reveals a dual role of CXCR4 in osteosarcoma primary tumor growth and metastasis. J Cancer Res Clin Oncol 142, 1739–1750 (2016). https://doi.org/10.1007/s00432-016-2185-5
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DOI: https://doi.org/10.1007/s00432-016-2185-5