PDGF-BB modulates hematopoiesis and tumor angiogenesis by inducing erythropoietin production in stromal cells

Abstract

The platelet-derived growth factor (PDGF) signaling system contributes to tumor angiogenesis and vascular remodeling. Here we show in mouse tumor models that PDGF-BB induces erythropoietin (EPO) mRNA and protein expression by targeting stromal and perivascular cells that express PDGF receptor-β (PDGFR-β). Tumor-derived PDGF-BB promoted tumor growth, angiogenesis and extramedullary hematopoiesis at least in part through modulation of EPO expression. Moreover, adenoviral delivery of PDGF-BB to tumor-free mice increased both EPO production and erythropoiesis, as well as protecting from irradiation-induced anemia. At the molecular level, we show that the PDGF-BB–PDGFR-bβ signaling system activates the EPO promoter, acting in part through transcriptional regulation by the transcription factor Atf3, possibly through its association with two additional transcription factors, c-Jun and Sp1. Our findings suggest that PDGF-BB–induced EPO promotes tumor growth through two mechanisms: first, paracrine stimulation of tumor angiogenesis by direct induction of endothelial cell proliferation, migration, sprouting and tube formation, and second, endocrine stimulation of extramedullary hematopoiesis leading to increased oxygen perfusion and protection against tumor-associated anemia.

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Figure 1: PDGF-BB in stromal expansion, angiogenesis and tumor growth.
Figure 2: Extramedullary hematopoiesis and expression of PDGFRs in the stromal compartment.
Figure 3: Elevation of plasma EPO concentrations and transcriptional regulation of EPO expression by PDGF-BB.
Figure 4: In vivo upregulation of Epo mRNA by PDGF-BB, co-localization of EPO protein with PDGFR-β+ structures and maintenance of EPO production in spleen by PDGFR-β.
Figure 5: Anti-tumor and antiangiogenic activity, systemic impact of EPO or PDGFR antagonism and the direct effects of EPO on endothelial cells.
Figure 6: AdPDGF-BB induces EPO expression, extramedullary hematopoiesis and increased hematocrit and improves irradiation-induced anemia.

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Acknowledgements

We thank J. Nissen and Z. Peng for their technical support. We thank Z. Zhu at ImClone for providing us the antibodies specific to mouse PDGFR-α and PDGFR-β. The MS-5 and S17 cell lines were provided by A. Berardi (Ospedale Bambin Gesu, Italy) and K. Dorshkind (University of California, Los Angeles, California, USA), and the adenoviruses were provided by S. Ylä-Herttuala (University of Kuopio, Kuopio, Finland). This work was supported by the laboratory of Y.C. through research grants from the Swedish Research Council, the Swedish Cancer Foundation, the Karolinska Institute Foundation, the Karolinska Institute distinguished professor award Torsten och Ragnar Söderbergs Stiftelser, a grant from ImClone, the European Union Integrated Project of Metoxia (project number 222741) and the European Research Council advanced grant ANGIOFAT (project number 250021).

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Y.C. designed the study and wrote the manuscript. Y.X., S.L., K.H., Z.W., L.D.E.J., R.C. and E.-M.H. performed mouse experiments, as well as histological and immunohistological analyses. Y.X. and S.L. measured the plasma EPO concentrations by ELISA, measured the luciferase activity, performed hematological analyses and performed adenoviral analyses. S.L. performed radiation experiments. Y.X., Z.W. and S.L. cultured stromal cells for in vitro assays. Y.Y. performed qRT-PCR, EMSA and ChIP assays. K.H. and Y.Y. performed FACS analyses. K.H. performed colony-forming cell assays and in vitro endothelial cell assays. P.A. performed the western blot analysis. S.L. prepared samples for in situ hybridization, and D.G. performed in situ hybridization assays. Y.X. prepared samples for the microarray assay. Y.X. and O.L. analyzed the microarray data. M.S. provided the PDGFR-β knockout mice for this study.

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Correspondence to Yihai Cao.

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Supplementary Methods, Supplementary Figures 1–7 and Supplementary Table 1 (PDF 1056 kb)

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Xue, Y., Lim, S., Yang, Y. et al. PDGF-BB modulates hematopoiesis and tumor angiogenesis by inducing erythropoietin production in stromal cells. Nat Med 18, 100–110 (2012). https://doi.org/10.1038/nm.2575

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