Advertisement

Molecular Medicine

, Volume 19, Issue 1, pp 223–229 | Cite as

Erythropoietin (EPO) Increases Myelin Gene Expression in CG4 Oligodendrocyte Cells through the Classical EPO Receptor

  • Ilaria Cervellini
  • Alexander Annenkov
  • Thomas Brenton
  • Yuti Chernajovsky
  • Pietro Ghezzi
  • Manuela Mengozzi
Research Article

Abstract

Erythropoietin (EPO) has protective effects in neurodegenerative and neuroinflammatory diseases, including in animal models of multiple sclerosis, where EPO decreases disease severity. EPO also promotes neurogenesis and is protective in models of toxic demyelination. In this study, we asked whether EPO could promote neurorepair by also inducing remyelination. In addition, we investigated whether the effect of EPO could be mediated by the classical erythropoietic EPO receptor (EPOR), since it is still questioned if EPOR is functional in nonhematopoietic cells. Using CG4 cells, a line of rat oligodendrocyte precursor cells, we found that EPO increases the expression of myelin genes (myelin oligodendrocyte glycoprotein (MOG) and myelin basic protein (MBP)). EPO had no effect in wild-type CG4 cells, which do not express EPOR, whereas it increased MOG and MBP expression in cells engineered to overexpress EPOR (CG4-EPOR). This was reflected in a marked increase in MOG protein levels, as detected by Western blot. In these cells, EPO induced by 10-fold the early growth response gene 2 (Egr2), which is required for peripheral myelination. However, Egr2 silencing with a siRNA did not reverse the effect of EPO, indicating that EPO acts through other pathways. In conclusion, EPO induces the expression of myelin genes in oligodendrocytes and this effect requires the presence of EPOR. This study demonstrates that EPOR can mediate neuroreparative effects.

Notes

Acknowledgments

PG is supported by the RM Phillips Charitable Trust and the European Regional Development Fund, TC2N. YC and AA are supported by NMSS-USA (Promise 2010).

References

  1. 1.
    Sargin D, Friedrichs H, El-Kordi A, Ehrenreich H. (2010) Erythropoietin as neuroprotective and neuroregenerative treatment strategy: comprehensive overview of 12 years of preclinical and clinical research. Best. Pract. Res. Clin. Anaesthesiol. 24:573–94.CrossRefPubMedGoogle Scholar
  2. 2.
    Bartels C, Spate K, Krampe H, Ehrenreich H. (2008) Recombinant human erythropoietin: novel strategies for neuroprotective/neuro-regenerative treatment of multiple sclerosis. Ther. Adv. Neurol. Disord. 1:193–206.CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Ghezzi P, Brines M. (2004) Erythropoietin as an antiapoptotic, tissue-protective cytokine. Cell Death Differ. 11 Suppl 1:S37–44.CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Wang L, Zhang Z, Wang Y, Zhang R, Chopp M. (2004) Treatment of stroke with erythropoietin enhances neurogenesis and angiogenesis and improves neurological function in rats. Stroke. 35:1732–7.CrossRefPubMedGoogle Scholar
  5. 5.
    Zhang L, et al. (2010) Erythropoietin amplifies stroke-induced oligodendrogenesis in the rat. PLoS One 5:e11016.CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Mengozzi M, et al. (2012) Erythropoietin-induced changes in brain gene expression reveal induction of synaptic plasticity genes in experimental stroke. Proc. Natl. Acad. Sci. U. S. A. 109:9617–22.CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Agnello D, et al. (2002) Erythropoietin exerts an anti-inflammatory effect on the CNS in a model of experimental autoimmune encephalomyelitis. Brain Res. 952:128–34.CrossRefPubMedGoogle Scholar
  8. 8.
    Savino C, et al. (2006) Delayed administration of erythropoietin and its non-erythropoietic derivatives ameliorates chronic murine autoimmune encephalomyelitis. J. Neuroimmunol. 172:27–37.CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Yuan R, et al. (2008) Erythropoietin: a potent inducer of peripheral immuno/inflammatory modulation in autoimmune EAE. PLoS ONE 3:e1924.CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Chen SJ, et al. (2010) Erythropoietin enhances endogenous haem oxygenase-1 and represses immune responses to ameliorate experimental autoimmune encephalomyelitis. Clin Exp Immunol 162:210–23.CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Li W, et al. (2004) Beneficial effect of erythropoietin on experimental allergic encephalomyelitis. Ann. Neurol. 56:767–77.CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Zhang J, et al. (2005) Erythropoietin treatment improves neurological functional recovery in EAE mice. Brain Res. 1034:34–9.CrossRefPubMedGoogle Scholar
  13. 13.
    Hagemeyer N, et al. (2012) Erythropoietin attenuates neurological and histological consequences of toxic demyelination in mice. Mol. Med. 18:628–35.CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Cho YK, et al. (2012) Erythropoietin promotes oligodendrogenesis and myelin repair following lysolecithin-induced injury in spinal cord slice culture. Biochem. Biophys. Res. Commun. 417:753–9.CrossRefPubMedGoogle Scholar
  15. 15.
    Sugawa M, Sakurai Y, Ishikawa-Ieda Y, Suzuki H, Asou H. (2002) Effects of erythropoietin on glial cell development; oligodendrocyte maturation and astrocyte proliferation. Neurosci. Res. 44:391–403.CrossRefPubMedGoogle Scholar
  16. 16.
    Ghezzi P, et al. (2010) Erythropoietin: not just about erythropoiesis. Lancet. 375:2142.CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Watowich SS, et al. (1992) Homodimerization and constitutive activation of the erythropoietin receptor. Proc. Natl. Acad. Sci. U. S. A. 89:2140–4.CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    Leist M, et al. (2004) Derivatives of erythropoietin that are tissue protective but not erythropoietic. Science. 305:239–42.CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Brines M, et al. (2008) Nonerythropoietic, tissue-protective peptides derived from the tertiary structure of erythropoietin. Proc. Natl. Acad. Sci. U. S. A. 105:10925–30.CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Xiong Y, et al. (2010) Erythropoietin improves histological and functional outcomes after traumatic brain injury in mice in the absence of the neural erythropoietin receptor. J. Neurotrauma. 27:205–15.CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Yu X, et al. (2002) Erythropoietin receptor signalling is required for normal brain development. Development. 129:505–16.PubMedGoogle Scholar
  22. 22.
    Um M, Gross AW, Lodish HF. (2007) A “classical” homodimeric erythropoietin receptor is essential for the antiapoptotic effects of erythropoietin on differentiated neuroblastoma SH-SY5Y and pheochromocytoma PC-12 cells. Cell Signal. 19:634–45.CrossRefPubMedGoogle Scholar
  23. 23.
    Siren AL, et al. (2001) Erythropoietin and erythropoietin receptor in human ischemic/hypoxic brain. Acta. Neuropathol. 101:271–6.PubMedGoogle Scholar
  24. 24.
    Brines M, Cerami A. (2012) The receptor that tames the innate immune response. Mol. Med. 18:486–96.CrossRefPubMedGoogle Scholar
  25. 25.
    Annenkov A, et al. (2011) A chimeric receptor of the insulin-like growth factor receptor type 1 (IGFR1) and a single chain antibody specific to myelin oligodendrocyte glycoprotein activates the IGF1R signalling cascade in CG4 oligoden-drocyte progenitors. Biochim. Biophys. Acta. 1813:1428–37.CrossRefPubMedGoogle Scholar
  26. 26.
    Wang M, Doucette JR, Nazarali AJ. (2011) Conditional Tet-regulated over-expression of Hoxa2 in CG4 cells increases their proliferation and delays their differentiation into oligodendrocyte-like cells expressing myelin basic protein. Cell Mol. Neurobiol. 31:875–86.CrossRefPubMedGoogle Scholar
  27. 27.
    Topilko P, et al. (1994) Krox-20 controls myelination in the peripheral nervous system. Nature. 371:796–9.CrossRefPubMedGoogle Scholar
  28. 28.
    Demaison C, et al. (2002) High-level transduction and gene expression in hematopoietic repopulating cells using a human imunodeficiency [immunodeficiency] virus type 1-based lentiviral vector containing an internal spleen focus forming virus promoter. Hum. Gene Ther. 13:803–13.CrossRefPubMedGoogle Scholar
  29. 29.
    Chen ZY, Wang L, Asavaritkrai P, Noguchi CT. (2010) Up-regulation of erythropoietin receptor by nitric oxide mediates hypoxia preconditioning. J. Neurosci. Res 88:3180–8.CrossRefPubMedGoogle Scholar
  30. 30.
    Zhang PL, et al. (2006) Increased myelinating capacity of embryonic stem cell derived oligodendrocyte precursors after treatment by interleukin-6/ soluble interleukin-6 receptor fusion protein. Mol. Cell. Neurosci. 31:387–98.CrossRefPubMedGoogle Scholar
  31. 31.
    Zhang PL, et al. (2007) Induction of neuronal and myelin-related gene expression by IL-6-receptor/IL-6: a study on embryonic dorsal root ganglia cells and isolated Schwann cells. Exp. Neurol. 208:285–96.CrossRefPubMedGoogle Scholar
  32. 32.
    Siren AL, et al. (2001) Erythropoietin prevents neuronal apoptosis after cerebral ischemia and metabolic stress. Proc. Natl. Acad. Sci. U. S. A. 98:4044–9.CrossRefPubMedPubMedCentralGoogle Scholar
  33. 33.
    Flores AI, et al. (2008) Constitutively active Akt induces enhanced myelination in the CNS. J. Neurosci. 28:7174–83.CrossRefPubMedPubMedCentralGoogle Scholar
  34. 34.
    Genc K, Genc S, Baskin H, Semin I. (2006) Erythropoietin decreases cytotoxicity and nitric oxide formation induced by inflammatory stimuli in rat oligodendrocytes. Physiol. Res. 55:33–8.PubMedGoogle Scholar
  35. 35.
    Kato S, et al. (2011) Endogenous erythropoietin from astrocyte protects the oligodendrocyte precursor cell against hypoxic and reoxygenation injury. J. Neurosci. Res 89:1566–74.CrossRefPubMedGoogle Scholar
  36. 36.
    Ehrenreich H, et al. (2007) Exploring recombinant human erythropoietin in chronic progressive multiple sclerosis. Brain. 130:2577–88.CrossRefPubMedPubMedCentralGoogle Scholar
  37. 37.
    Iwai M, et al. (2010) Enhanced oligodendrogenesis and recovery of neurological function by erythropoietin after neonatal hypoxic/ischemic brain injury. Stroke. 41:1032–7.CrossRefPubMedPubMedCentralGoogle Scholar
  38. 38.
    Yamada M, Burke C, Colditz P, Johnson DW, Gobe GC. (2011) Erythropoietin protects against apoptosis and increases expression of nonneuronal cell markers in the hypoxia-injured developing brain. J. Pathol. 224:101–9.CrossRefPubMedGoogle Scholar
  39. 39.
    McIver SR, et al. (2010) Oligodendrocyte degeneration and recovery after focal cerebral ischemia. Neuroscience. 169:1364–75.CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© The Author(s) 2013

Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, and provide a link to the Creative Commons license. You do not have permission under this license to share adapted material derived from this article or parts of it.

The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder.

To view a copy of this license, visit (https://doi.org/creativecommons.org/licenses/by-nc-nd/4.0/)

Authors and Affiliations

  • Ilaria Cervellini
    • 1
  • Alexander Annenkov
    • 2
  • Thomas Brenton
    • 1
  • Yuti Chernajovsky
    • 2
  • Pietro Ghezzi
    • 1
  • Manuela Mengozzi
    • 1
  1. 1.Trafford Centre for Medical ResearchBrighton and Sussex Medical SchoolBrightonUK
  2. 2.Bone and Joint Research Unit, William Harvey Research Institute, Bart’s and the London School of MedicineQueen Mary University of LondonLondonUK

Personalised recommendations