Epo's erythropoietic capacity is ascribed largely to its antiapoptotic actions. In part via gene profiling of bone marrow erythroblasts, Epo is now shown to selectively down-modulate the adhesion/migration factors chemokine receptor-4 (Cxcr4) and integrin alpha-4 (Itga4) and to up-modulate growth differentiation factor-3 (Gdf3), oncostatin-M (OncoM), and podocalyxin like-1 (PODXL). For PODXL, Epo dose–dependent expression of this CD34-related sialomucin was discovered in Kit^CD71^{proerythroblasts and was sustained at subsequent Kit}^CD71^{and Ter119}^{stages. In vivo, Epo markedly induced PODXL expression in these progenitors and in marrow-resident reticulocytes. This was further associated with a rapid release of PODXL}^{reticulocytes to blood. As studied in erythroblasts expressing minimal Epo receptor (EpoR) alleles, efficient PODXL induction proved dependence on an EpoR-PY343 Stat5 binding site. Moreover, in mice expressing an EpoR-HM F343 allele, compromised Epo-induced PODXL expression correlated with abnormal anucleated red cell representation in marrow. By modulating this select set of cell-surface adhesion molecules and chemokines, Epo is proposed to mobilize erythroblasts from a hypothesized stromal niche and possibly promote reticulocyte egress to blood.}

Abstract

Values are percentages.

Click here to view.

Acknowledgments

The authors thank A. Fiser and K. Miller for expert assistance in manuscript assembly.

This work was supported by National Institutes of Health (NIH) R01 HL044491 and core facilities sponsored by NIH NCRR COBRE grants P20-RR18789 and P20-RR15555.

Acknowledgments

Footnotes

The online version of this article contains a data supplement.

The publication costs of this article were defrayed in part by page charge payment. Therefore, and solely to indicate this fact, this article is hereby marked “advertisement” in accordance with 18 USC section 1734.

Footnotes

References

1. Baumann R, Dragon SErythropoiesis and red cell function in vertebrate embryos. Eur J Clin Invest. 2005;35(suppl 3):2–12.[PubMed][Google Scholar]
2. Cantor AB, Orkin SHCoregulation of GATA factors by the Friend of GATA (FOG) family of multitype zinc finger proteins. Semin Cell Dev Biol. 2005;16:117–128.[PubMed][Google Scholar]
3. Richmond TD, Chohan M, Barber DLTurning cells red: signal transduction mediated by erythropoietin. Trends Cell Biol. 2005;15:146–155.[PubMed][Google Scholar]
4. Koury MJErythropoietin: the story of hypoxia and a finely regulated hematopoietic hormone. Exp Hematol. 2005;33:1263–1270.[PubMed][Google Scholar]
5. Koury MJ, Bondurant MCErythropoietin retards DNA breakdown and prevents programmed death in erythroid progenitor cells. Science. 1990;248:378–381.[PubMed][Google Scholar]
6. Wu H, Liu X, Jaenisch R, Lodish HFGeneration of committed erythroid BFU-E and CFU-E progenitors does not require erythropoietin or the erythropoietin receptor. Cell. 1995;83:59–67.[PubMed][Google Scholar]
7. Socolovsky M, Fallon AE, Wang S, Brugnara C, Lodish HFFetal anemia and apoptosis of red cell progenitors in Stat5a−/−5b−/− mice: a direct role for Stat5 in Bcl-X(L) induction. Cell. 1999;98:181–191.[PubMed][Google Scholar]
8. Ghaffari S, Kitidis C, Zhao W, et al AKT induces erythroid-cell maturation of JAK2-deficient fetal liver progenitor cells and is required for Epo regulation of erythroid-cell differentiation. Blood. 2006;107:1888–1891.[Google Scholar]
9. Levine AJ, Feng Z, Mak TW, You H, Jin SCoordination and communication between the p53 and IGF-1-AKT-TOR signal transduction pathways. Genes Dev. 2006;20:267–275.[PubMed][Google Scholar]
10. Hammerman PS, Fox CJ, Birnbaum MJ, Thompson CBPim and Akt oncogenes are independent regulators of hematopoietic cell growth and survival. Blood. 2005;105:4477–4483.[Google Scholar]
11. Maiese K, Li F, Chong ZZNew avenues of exploration for erythropoietin. JAMA. 2005;293:90–95.[Google Scholar]
12. Menon MP, Karur V, Bogacheva O, Bogachev O, Cuetara B, Wojchowski DMSignals for stress erythropoiesis are integrated via an erythropoietin receptor-phosphotyrosine-343-Stat5 axis. J Clin Invest. 2006;116:683–694.[Google Scholar]
13. Levine AJ, Brivanlou AHGDF3 at the crossroads of TGF-beta signaling. Cell Cycle. 2006;5:1069–1073.[PubMed][Google Scholar]
14. Tanaka M, Miyajima AOncostatin M, a multifunctional cytokine. Rev Physiol Biochem Pharmacol. 2003;149:39–52.[PubMed][Google Scholar]
15. Kucia M, Reca R, Miekus K, et al Trafficking of normal stem cells and metastasis of cancer stem cells involve similar mechanisms: pivotal role of the SDF-1-CXCR4 axis. Stem Cells. 2005;23:879–894.[PubMed][Google Scholar]
16. Scott LM, Priestley GV, Papayannopoulou TDeletion of alpha4 integrins from adult hematopoietic cells reveals roles in homeostasis, regeneration, and homing. Mol Cell Biol. 2003;23:9349–9360.[Google Scholar]
17. Arroyo AG, Yang JT, Rayburn H, Hynes ROAlpha4 integrins regulate the proliferation/differentiation balance of multilineage hematopoietic progenitors in vivo. Immunity. 1999;11:555–566.[PubMed][Google Scholar]
18. Barton Furness SG, McNagny KBeyond mere markers: functions for CD34 family of sialomucins in hematopoiesis. Immunol Res. 2006;34:13–32.[PubMed][Google Scholar]
19. Tanaka M, Hirabayashi Y, Sekiguchi T, Inoue T, Katsuki M, Miyajima ATargeted disruption of oncostatin M receptor results in altered hematopoiesis. Blood. 2003;102:3154–3162.[PubMed][Google Scholar]
20. Jorcyk CL, Holzer RG, Ryan REOncostatin M induces cell detachment and enhances the metastatic capacity of T-47D human breast carcinoma cells. Cytokine. 2006;33:323–336.[PubMed][Google Scholar]
21. McPherron AC, Lee SJGDF-3 and GDF-9: two new members of the transforming growth factor-beta superfamily containing a novel pattern of cysteines. J Biol Chem. 1993;268:3444–3449.[PubMed][Google Scholar]
22. Elisseeva EL, Slupsky CM, Crump MP, Clark-Lewis I, Sykes BDNMR studies of active N-terminal peptides of stromal cell-derived factor-1: structural basis for receptor binding. J Biol Chem. 2000;275:26799–26805.[PubMed][Google Scholar]
23. Peterson JA, Sheibani N, David G, Garcia-Pardo A, Peters DMHeparin II domain of fibronectin uses alpha4beta1 integrin to control focal adhesion and stress fiber formation, independent of syndecan-4. J Biol Chem. 2005;280:6915–6922.[PubMed][Google Scholar]
24. Rose DMThe role of the alpha4 integrin-paxillin interaction in regulating leukocyte trafficking. Exp Mol Med. 2006;38:191–195.[PubMed][Google Scholar]
25. Dekan G, Gabel C, Farquhar MGSulfate contributes to the negative charge of podocalyxin, the major sialoglycoprotein of the glomerular filtration slits. Proc Natl Acad Sci U S A. 1991;88:5398–5402.[Google Scholar]
26. Doyonnas R, Nielsen JS, Chelliah S, et al Podocalyxin is a CD34-related marker of murine hematopoietic stem cells and embryonic erythroid cells. Blood. 2005;105:4170–4178.[PubMed][Google Scholar]
27. Wilson A, Trumpp ABone-marrow haematopoietic-stem-cell niches. Nat Rev Immunol. 2006;6:93–106.[PubMed][Google Scholar]
28. Nagasawa TMicroenvironmental niches in the bone marrow required for B-cell development. Nat Rev Immunol. 2006;6:107–116.[PubMed][Google Scholar]
29. Chasis JAErythroblastic islands: specialized microenvironmental niches for erythropoiesis. Curr Opin Hematol. 2006;13:137–141.[PubMed][Google Scholar]
30. Lee G, Lo A, Short SA, et al Targeted gene deletion demonstrates that cell adhesion molecule ICAM-4 is critical for erythroblastic island formation. Blood. 2006;108:2064–2071.[Google Scholar]
31. Zang H, Sato K, Nakajima H, McKay C, Ney PA, Ihle JNThe distal region and receptor tyrosines of the Epo receptor are nonessential for in vivo erythropoiesis. EMBO J. 2001;20:3156–3166.[Google Scholar]
32. Wojchowski DM, Caslake LBiotinylated recombinant human erythropoietins: bioactivity and utility as receptor ligand. Blood. 1989;74:952–958.[PubMed][Google Scholar]
33. Menon MP, Fang J, Wojchowski DMCore erythropoietin receptor signals for late erythroblast development. Blood. 2006;107:2662–2672.[Google Scholar]
34. Matsumoto A, Masuhara M, Mitsui K, et al CIS, a cytokine inducible SH2 protein, is a target of the JAK-STAT5 pathway and modulates STAT5 activation. Blood. 1997;89:3148–3154.[PubMed][Google Scholar]
35. Son BR, Marquez-Curtis LA, Kucia M, et al Migration of bone marrow and cord blood mesenchymal stem cells in vitro is regulated by stromal-derived factor-1-CXCR4 and hepatocyte growth factor-c-met axes and involves matrix metalloproteinases. Stem Cells. 2006;24:1254–1264.[PubMed][Google Scholar]
36. Karur VG, Lowell CA, Besmer P, Agosti V, Wojchowski DMLyn kinase promotes erythroblast expansion and late-stage development. Blood. 2006;108:1524–1532.[Google Scholar]
37. Wijelath ES, Carlsen B, Cole T, Chen J, Kothari S, Hammond WPOncostatin M induces basic fibroblast growth factor expression in endothelial cells and promotes endothelial cell proliferation migration and spindle morphology. J Cell Sci. 1997;110(pt 7):871–879.[PubMed][Google Scholar]
38. Strand K, Murray J, Aziz S, et al Induction of the urokinase plasminogen activator system by oncostatin M promotes endothelial migration. J Cell Biochem. 2000;79:239–248.[PubMed][Google Scholar]
39. Modur V, Feldhaus MJ, Weyrich AS, et al Oncostatin M is a proinflammatory mediator: in vivo effects correlate with endothelial cell expression of inflammatory cytokines and adhesion molecules. J Clin Invest. 1997;100:158–168.[Google Scholar]
40. Zermati Y, Fichelson S, Valensi F, et al Transforming growth factor inhibits erythropoiesis by blocking proliferation and accelerating differentiation of erythroid progenitors. Exp Hematol. 2000;28:885–894.[PubMed][Google Scholar]
41. Lenox LE, Perry JM, Paulson RFBMP4 and Madh5 regulate the erythroid response to acute anemia. Blood. 2005;105:2741–2748.[PubMed][Google Scholar]
42. Ghio M, Ottonello L, Contini P, et al Transforming growth factor-beta1 in supernatants from stored red blood cells inhibits neutrophil locomotion. Blood. 2003;102:1100–1107.[PubMed][Google Scholar]
43. Imitola J, Raddassi K, Park KI, et al Directed migration of neural stem cells to sites of CNS injury by the stromal cell-derived factor 1alpha/CXC chemokine receptor 4 pathway. Proc Natl Acad Sci U S A. 2004;101:18117–18122.[Google Scholar]
44. Zernecke A, Schober A, Bot I, et al SDF-1alpha/CXCR4 axis is instrumental in neointimal hyperplasia and recruitment of smooth muscle progenitor cells. Circ Res. 2005;96:784–791.[PubMed][Google Scholar]
45. Belmadani A, Tran PB, Ren D, Assimacopoulos S, Grove EA, Miller RJThe chemokine stromal cell-derived factor-1 regulates the migration of sensory neuron progenitors. J Neurosci. 2005;25:3995–4003.[Google Scholar]
46. Kucia M, Jankowski K, Reca R, et al CXCR4-SDF-1 signalling, locomotion, chemotaxis and adhesion. J Mol Histol. 2004;35:233–245.[PubMed][Google Scholar]
47. Simmons PJ, Leavesley DI, Levesque JP, et al The mobilization of primitive hemopoietic progenitors into the peripheral blood. Stem Cells. 1994;2(suppl 1):187–201. discussion 201–182. [[PubMed][Google Scholar]
48. Vitelli L, Condorelli G, Lulli V, et al A pentamer transcriptional complex including tal-1 and retinoblastoma protein downmodulates c-kit expression in normal erythroblasts. Mol Cell Biol. 2000;20:5330–5342.[Google Scholar]
49. Felli N, Fontana L, Pelosi E, et al MicroRNAs 221 and 222 inhibit normal erythropoiesis and erythroleukemic cell growth via kit receptor down-modulation. Proc Natl Acad Sci U S A. 2005;102:18081–18086.[Google Scholar]
50. Kim HK, De La Luz Sierra M, Williams CK, Gulino AV, Tosato GG-CSF down-regulation of CXCR4 expression identified as a mechanism for mobilization of myeloid cells. Blood. 2006;108:812–820.[Google Scholar]
51. Oostendorp RA, Dormer PVLA-4-mediated interactions between normal human hematopoietic progenitors and stromal cells. Leuk Lymphoma. 1997;24:423–435.[PubMed][Google Scholar]
52. Barber DL, Beattie BK, Mason JM, et al A common epitope is shared by activated signal transducer and activator of transcription-5 (STAT5) and the phosphorylated erythropoietin receptor: implications for the docking model of STAT activation. Blood. 2001;97:2230–2237.[PubMed][Google Scholar]
53. Gobert S, Chretien S, Gouilleux F, et al Identification of tyrosine residues within the intracellular domain of the erythropoietin receptor crucial for STAT5 activation. EMBO J. 1996;15:2434–2441.[Google Scholar]
54. Palmer RE, Kotsianti A, Cadman B, Boyd T, Gerald W, Haber DAWT1 regulates the expression of the major glomerular podocyte membrane protein Podocalyxin. Curr Biol. 2001;11:1805–1809.[PubMed][Google Scholar]
55. Stanhope-Baker P, Kessler PM, Li W, Agarwal ML, Williams BRThe Wilms tumor suppressor-1 target gene podocalyxin is transcriptionally repressed by p53. J Biol Chem. 2004;279:33575–33585.[PubMed][Google Scholar]
56. Birkenkamp KU, Geugien M, Lemmink HH, Kruijer W, Vellenga ERegulation of constitutive STAT5 phosphorylation in acute myeloid leukemia blasts. Leukemia. 2001;15:1923–1931.[PubMed][Google Scholar]
57. Spiekermann K, Pau M, Schwab R, Schmieja K, Franzrahe S, Hiddemann WConstitutive activation of STAT3 and STAT5 is induced by leukemic fusion proteins with protein tyrosine kinase activity and is sufficient for transformation of hematopoietic precursor cells. Exp Hematol. 2002;30:262–271.[PubMed][Google Scholar]
58. Barash IStat5 in the mammary gland: controlling normal development and cancer. J Cell Physiol. 2006;209:305–313.[PubMed][Google Scholar]
59. Nevalainen MT, Xie J, Torhorst J, et al Signal transducer and activator of transcription-5 activation and breast cancer prognosis. J Clin Oncol. 2004;22:2053–2060.[PubMed][Google Scholar]
60. Kazansky AV, Spencer DM, Greenberg NMActivation of signal transducer and activator of transcription 5 is required for progression of autochthonous prostate cancer: evidence from the transgenic adenocarcinoma of the mouse prostate system. Cancer Res. 2003;63:8757–8762.[PubMed][Google Scholar]
61. Li H, Ahonen TJ, Alanen K, et al Activation of signal transducer and activator of transcription 5 in human prostate cancer is associated with high histologic grade. Cancer Res. 2004;64:4774–4782.[PubMed][Google Scholar]
62. Kelley TW, Huntsman D, McNagny KM, Roskelley CD, Hsi EDPodocalyxin: a marker of blasts in acute leukemia. Am J Clin Pathol. 2005;124:134–142.[PubMed][Google Scholar]
63. Somasiri A, Nielsen JS, Makretsov N, et al Overexpression of the antiadhesin podocalyxin is an independent predictor of breast cancer progression. Cancer Res. 2004;64:5068–5073.[PubMed][Google Scholar]
64. Casey G, Neville PJ, Liu X, et al Podocalyxin variants and risk of prostate cancer and tumor aggressiveness. Hum Mol Genet. 2006;15:735–741.[PubMed][Google Scholar]
65. Leyland-Jones BBreast cancer trial with erythropoietin terminated unexpectedly. Lancet Oncol. 2003;4:459–460.[PubMed][Google Scholar]
66. Henke M, Laszig R, Rube C, et al Erythropoietin to treat head and neck cancer patients with anaemia undergoing radiotherapy: randomised, double-blind, placebo-controlled trial. Lancet. 2003;362:1255–1260.[PubMed][Google Scholar]
67. Feldman L, Wang Y, Rhim JS, Bhattacharya N, Loda M, Sytkowski AJErythropoietin stimulates growth and STAT5 phosphorylation in human prostate epithelial and prostate cancer cells. Prostate. 2006;66:135–145.[PubMed][Google Scholar]
68. Li K, Menon MP, Karur VG, Hegde S, Wojchowski DMAttenuated signaling by a phosphotyrosine-null Epo receptor form in primary erythroid progenitor cells. Blood. 2003;102:3147–3153.[PubMed][Google Scholar]
69. Tusher VG, Tibshirani R, Chu GSignificance analysis of microarrays applied to the ionizing radiation response. Proc Natl Acad Sci U S A. 2001;98:5116–5121.[Google Scholar]
70. Storey JD, Tibshirani RStatistical methods for identifying differentially expressed genes in DNA microarrays. Methods Mol Biol. 2003;224:149–157.[PubMed][Google Scholar]