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

Nature Medicine volume 18, pages 100110 (2012) | Download Citation

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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References

  1. 1.

    Molecular mechanisms and therapeutic development of angiogenesis inhibitors. Adv. Cancer Res. 100, 113–131 (2008).

  2. 2.

    Angiogenesis in life, disease and medicine. Nature 438, 932–936 (2005).

  3. 3.

    Angiogenesis: an organizing principle for drug discovery? Nat. Rev. Drug Discov. 6, 273–286 (2007).

  4. 4.

    Tumor angiogenesis. N. Engl. J. Med. 358, 2039–2049 (2008).

  5. 5.

    , & Endothelial and nonendothelial sources of PDGF-B regulate pericyte recruitment and influence vascular pattern formation in tumors. J. Clin. Invest. 112, 1142–1151 (2003).

  6. 6.

    & Angiogenesis as a therapeutic target. Nature 438, 967–974 (2005).

  7. 7.

    , , & High interstitial fluid pressure—an obstacle in cancer therapy. Nat. Rev. Cancer 4, 806–813 (2004).

  8. 8.

    , , & Basic fibroblast growth factor secreted by an animal tumor is detectable in urine. Cancer Res. 53, 5297–5299 (1993).

  9. 9.

    et al. Angiopoietin-1 protects the adult vasculature against plasma leakage. Nat. Med. 6, 460–463 (2000).

  10. 10.

    , & Growth factors and oncogenes in human malignant glioma. Neurol. Clin. 3, 785–799 (1985).

  11. 11.

    Positive and negative modulation of angiogenesis by VEGFR1 ligands. Sci. Signal. 2, re1 (2009).

  12. 12.

    et al. Anti-VEGF agents confer survival advantages to tumor-bearing mice by improving cancer-associated systemic syndrome. Proc. Natl. Acad. Sci. USA 105, 18513–18518 (2008).

  13. 13.

    et al. Angiogenic factors FGF2 and PDGF-BB synergistically promote murine tumor neovascularization and metastasis. J. Clin. Invest. 117, 2766–2777 (2007).

  14. 14.

    & Modes of resistance to anti-angiogenic therapy. Nat. Rev. Cancer 8, 592–603 (2008).

  15. 15.

    et al. PDGF-C mediates the angiogenic and tumorigenic properties of fibroblasts associated with tumors refractory to anti-VEGF treatment. Cancer Cell 15, 21–34 (2009).

  16. 16.

    & Tumor-stroma interactions. Curr. Opin. Genet. Dev. 15, 97–101 (2005).

  17. 17.

    & Fibroblasts in cancer. Nat. Rev. Cancer 6, 392–401 (2006).

  18. 18.

    , , , & PDGF receptors as cancer drug targets. Cancer Cell 3, 439–443 (2003).

  19. 19.

    et al. Tie2 identifies a hematopoietic lineage of proangiogenic monocytes required for tumor vessel formation and a mesenchymal population of pericyte progenitors. Cancer Cell 8, 211–226 (2005).

  20. 20.

    & Structure and function of platelet-derived growth factor. Acta Med. Scand. Suppl. 715, 19–23 (1987).

  21. 21.

    et al. Angiogenesis stimulated by PDGF-CC, a novel member in the PDGF family, involves activation of PDGFR-αα and -αβ receptors. FASEB J. 16, 1575–1583 (2002).

  22. 22.

    et al. Angiogenic synergism, vascular stability and improvement of hind-limb ischemia by a combination of PDGF-BB and FGF-2. Nat. Med. 9, 604–613 (2003).

  23. 23.

    et al. Differential roles of PDGFR-α and PDGFR-β in angiogenesis and vessel stability. FASEB J. 23, 153–163 (2009).

  24. 24.

    , , & Pericyte loss and microaneurysm formation in PDGF-B–deficient mice. Science 277, 242–245 (1997).

  25. 25.

    et al. Paracrine PDGF-B/PDGF-Rβ signaling controls mesangial cell development in kidney glomeruli. Development 125, 3313–3322 (1998).

  26. 26.

    Abnormal kidney development and hematological disorders in PDGF β-receptor mutant mice. Genes Dev. 8, 1888–1896 (1994).

  27. 27.

    , & Developmental regulation of erythropoietin and erythropoiesis. Am. J. Physiol. 273, R1829–R1844 (1997).

  28. 28.

    Pleiotropic functions of erythropoietin. Intern. Med. 42, 142–149 (2003).

  29. 29.

    , , , & Erythropoietin as an angiogenic factor. Eur. J. Clin. Invest. 33, 891–896 (2003).

  30. 30.

    et al. The relevance of extramedullary hemopoiesis to the staging of chronic myeloid leukemia. Boll. Ist. Sieroter. Milan. 57, 257–270 (1978).

  31. 31.

    , , & Idiopathic refractory sideroblastic anemia: clinical and laboratory investigation of 17 patients and review of the literature. Medicine (Baltimore) 50, 139–159 (1971).

  32. 32.

    et al. Erythropoietin and erythropoietin receptor coexpression is associated with poor survival in stage I non-small cell lung cancer. Clin. Cancer Res. 13, 4825–4831 (2007).

  33. 33.

    , , & Extramedullary erythropoiesis in the adult liver requires BMP-4/Smad5-dependent signaling. Exp. Hematol. 37, 549–558 (2009).

  34. 34.

    , & Effect of fibroblast growth factor 2 on stromal cell-derived factor 1 production by bone marrow stromal cells and hematopoiesis. J. Natl. Cancer Inst. 99, 223–235 (2007).

  35. 35.

    et al. Neuronal injury-inducible gene is synergistically regulated by ATF3, c-Jun, and STAT3 through the interaction with Sp1 in damaged neurons. J. Biol. Chem. 283, 6988–6996 (2008).

  36. 36.

    et al. PDGF receptor β is a potent regulator of mesenchymal stromal cell function. J. Bone Miner. Res. 23, 1519–1528 (2008).

  37. 37.

    et al. HIF-1–induced erythropoietin in the hypoxic retina protects against light-induced retinal degeneration. Nat. Med. 8, 718–724 (2002).

  38. 38.

    et al. Erythropoietin blockade inhibits the induction of tumor angiogenesis and progression. PLoS ONE 2, e549 (2007).

  39. 39.

    et al. Vascular endothelial growth factor-A and platelet-derived growth factor-B combination gene therapy prolongs angiogenic effects via recruitment of interstitial mononuclear cells and paracrine effects rather than improved pericyte coverage of angiogenic vessels. Circ. Res. 103, 1092–1099 (2008).

  40. 40.

    , , , & AMPKα1 deletion shortens erythrocyte life span in mice: role of oxidative stress. J. Biol. Chem. 285, 19976–19985 (2010).

  41. 41.

    et al. Erythropoietin stimulates spleen BMP4-dependent stress erythropoiesis and partially corrects anemia in a mouse model of generalized inflammation. Blood 116, 6072–6081 (2010).

  42. 42.

    , , , & Murine erythroid short-term radioprotection requires a BMP4-dependent, self-renewing population of stress erythroid progenitors. J. Clin. Invest. 120, 4507–4519 (2010).

  43. 43.

    et al. Induction of activating transcription factor 3 by anoxia is independent of p53 and the hypoxic HIF signalling pathway. Oncogene 26, 284–289 (2007).

  44. 44.

    , , & Erythropoietin deficiency decreases vascular stability in mice. J. Clin. Invest. 118, 526–533 (2008).

  45. 45.

    et al. Erythropoietin regulates endothelial progenitor cells. Blood 103, 921–926 (2004).

  46. 46.

    , , , & Plasma platelet-derived growth factor-B chain is elevated in patients with extensively large brain tumour. Acta Neurochir. (Wien) 137, 182–187 (1995).

  47. 47.

    et al. Elevated plasma platelet-derived growth factor B-chain levels in cancer patients. Cancer Res. 51, 4149–4154 (1991).

  48. 48.

    et al. Renal cell carcinoma in a patient with Beckwith-Wiedemann syndrome. Pediatr. Radiol. 26, 312–314 (1996).

  49. 49.

    et al. Soluble receptor-mediated selective inhibition of VEGFR and PDGFR-β signaling during physiologic and tumor angiogenesis. Proc. Natl. Acad. Sci. USA 105, 10185–10190 (2008).

  50. 50.

    , & The absence of pericytes does not increase the sensitivity of tumor vasculature to vascular endothelial growth factor-A blockade. Cancer Res. 70, 5109–5115 (2010).

  51. 51.

    & Erythropoietin promotes resistance against the Abl tyrosine kinase inhibitor imatinib (STI571) in K562 human leukemia cells. Mol. Cancer Res. 1, 970–980 (2003).

  52. 52.

    et al. Hypoxia-independent angiogenesis in adipose tissues during cold acclimation. Cell Metab. 9, 99–109 (2009).

  53. 53.

    et al. PDGF-BB induces intratumoral lymphangiogenesis and promotes lymphatic metastasis. Cancer Cell 6, 333–345 (2004).

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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).

Author information

Affiliations

  1. Department of Microbiology, Tumor and Cell Biology, Karolinska Institute, Stockholm, Sweden.

    • Yuan Xue
    • , Sharon Lim
    • , Yunlong Yang
    • , Zongwei Wang
    • , Lasse Dahl Ejby Jensen
    • , Eva-Maria Hedlund
    • , Patrik Andersson
    • , Renhai Cao
    • , Kayoko Hosaka
    •  & Yihai Cao
  2. Department of Pathology, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, Toyama, Japan.

    • Masakiyo Sasahara
  3. Department of Oncology-Pathology, Karolinska Institute, Stockholm, Sweden.

    • Ola Larsson
  4. Department of Neuroscience, Karolinska Institute, Stockholm, Sweden.

    • Dagmar Galter
  5. Department of Medicine and Health Sciences, Linköping University, Linköping, Sweden.

    • Yihai Cao

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Contributions

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.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Yihai Cao.

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https://doi.org/10.1038/nm.2575

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