Abstract
The pathophysiology of ineffective erythropoiesis in β-thalassemia is poorly understood. We report that RAP-011, an activin receptor IIA (ActRIIA) ligand trap, improved ineffective erythropoiesis, corrected anemia and limited iron overload in a mouse model of β-thalassemia intermedia. Expression of growth differentiation factor 11 (GDF11), an ActRIIA ligand, was increased in splenic erythroblasts from thalassemic mice and in erythroblasts and sera from subjects with β-thalassemia. Inactivation of GDF11 decreased oxidative stress and the amount of α-globin membrane precipitates, resulting in increased terminal erythroid differentiation. Abnormal GDF11 expression was dependent on reactive oxygen species, suggesting the existence of an autocrine amplification loop in β-thalassemia. GDF11 inactivation also corrected the abnormal ratio of immature/mature erythroblasts by inducing apoptosis of immature erythroblasts through the Fas–Fas ligand pathway. Taken together, these observations suggest that ActRIIA ligand traps may have therapeutic relevance in β-thalassemia by suppressing the deleterious effects of GDF11, a cytokine which blocks terminal erythroid maturation through an autocrine amplification loop involving oxidative stress and α-globin precipitation.
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References
Higgs, D.R., Engel, J.D. & Stamatoyannopoulos, G. Thalassaemia. Lancet 379, 373–383 (2012).
Weiss, M.J. & dos Santos, C.O. Chaperoning erythropoiesis. Blood 113, 2136–2144 (2009).
Kihm, A.J. et al. An abundant erythroid protein that stabilizes free α-haemoglobin. Nature 417, 758–763 (2002).
Ribeil, J.A. et al. Ineffective erythropoiesis in β-thalassemia. ScientificWorldJournal 2013, 1–11 (2013).
Sorensen, S., Rubin, E., Polster, H., Mohandas, N. & Schrier, S. The role of membrane skeletal-associated α-globin in the pathophysiology of β-thalassemia. Blood 75, 1333–1336 (1990).
Ginzburg, Y. & Rivella, S. β-thalassemia: a model for elucidating the dynamic regulation of ineffective erythropoiesis and iron metabolism. Blood 118, 4321–4330 (2011).
Zermati, Y. et al. Transforming growth factor inhibits erythropoiesis by blocking proliferation and accelerating differentiation of erythroid progenitors. Exp. Hematol. 28, 885–894 (2000).
Chadwick, K. et al. Cytokines and BMP-4 promote hematopoietic differentiation of human embryonic stem cells. Blood 102, 906–915 (2003).
Hegde, S. et al. An intronic sequence mutated in flexed-tail mice regulates splicing of Smad5. Mamm. Genome 18, 852–860 (2007).
Paulson, R.F., Shi, L. & Wu, D.C. Stress erythropoiesis: new signals and new stress progenitor cells. Curr. Opin. Hematol. 18, 139–145 (2011).
Tanno, T. et al. High levels of GDF15 in thalassemia suppress expression of the iron regulatory protein hepcidin. Nat. Med. 13, 1096–1101 (2007).
Broxmeyer, H.E. et al. Selective and indirect modulation of human multipotential and erythroid hematopoietic progenitor cell proliferation by recombinant human activin and inhibin. Proc. Natl. Acad. Sci. USA 85, 9052–9056 (1988).
Nakao, K., Kosaka, M. & Saito, S. Effects of erythroid differentiation factor (EDF) on proliferation and differentiation of human hematopoietic progenitors. Exp. Hematol. 19, 1090–1095 (1991).
Mizuguchi, T., Kosaka, M. & Saito, S. Activin A suppresses proliferation of interleukin-3-responsive granulocyte-macrophage colony-forming progenitors and stimulates proliferation and differentiation of interleukin-3-responsive erythroid burst-forming progenitors in the peripheral blood. Blood 81, 2891–2897 (1993).
Shiozaki, M., Kosaka, M. & Eto, Y. Activin A: a commitment factor in erythroid differentiation. Biochem. Biophys. Res. Commun. 242, 631–635 (1998).
Yu, J. et al. Importance of FSH-releasing protein and inhibin in erythrodifferentiation. Nature 330, 765–767 (1987).
Ruckle, J. et al. Single-dose, randomized, double-blind, placebo-controlled study of ACE-011 (ActRIIA-IgG1) in postmenopausal women. J. Bone Miner. Res. 24, 744–752 (2009).
Skow, L.C. et al. A mouse model for β-thalassemia. Cell 34, 1043–1052 (1983).
Ramos, P. et al. Iron metabolism and ineffective erythropoiesis in β-thalassemia mouse models. Ann. NY Acad. Sci. 1202, 24–30 (2010).
Liu, Y. et al. Suppression of Fas-FasL coexpression by erythropoietin mediates erythroblast expansion during the erythropoietic stress response in vivo. Blood 108, 123–133 (2006).
Mathias, L.A. et al. Ineffective erythropoiesis in β-thalassemia major is due to apoptosis at the polychromatophilic normoblast stage. Exp. Hematol. 28, 1343–1353 (2000).
Rivella, S. The role of ineffective erythropoiesis in non-transfusion-dependent thalassemia. Blood Rev. 26 (suppl. 1), S12–S15 (2012).
Tanno, T., Noel, P. & Miller, J.L. Growth differentiation factor 15 in erythroid health and disease. Curr. Opin. Hematol. 17, 184–190 (2010).
Wakefield, L.M. & Hill, C.S. Beyond TGFβ: roles of other TGFβ superfamily members in cancer. Nat. Rev. Cancer 13, 328–341 (2013).
Fainsod, A. et al. The dorsalizing and neural inducing gene follistatin is an antagonist of BMP-4. Mech. Dev. 63, 39–50 (1997).
Re'em-Kalma, Y., Lamb, T. & Frank, D. Competition between noggin and bone morphogenetic protein 4 activities may regulate dorsalization during Xenopus development. Proc. Natl. Acad. Sci. USA 92, 12141–12145 (1995).
Coulon, S. et al. Polymeric IgA1 controls erythroblast proliferation and accelerates erythropoiesis recovery in anemia. Nat. Med. 17, 1456–1465 (2011).
Iancu-Rubin, C. et al. Stromal cell-mediated inhibition of erythropoiesis can be attenuated by sotatercept (ACE-011), an activin receptor type II ligand trap. Exp. Hematol. 41, 155–166 (2013).
Yu, X. et al. An erythroid chaperone that facilitates folding of α-globin subunits for hemoglobin synthesis. J. Clin. Invest. 117, 1856–1865 (2007).
Marinkovic, D. et al. Foxo3 is required for the regulation of oxidative stress in erythropoiesis. J. Clin. Invest. 117, 2133–2144 (2007).
Suragani, R.N. et al. Heme-regulated eIF2α kinase activated Atf4 signaling pathway in oxidative stress and erythropoiesis. Blood 119, 5276–5284 (2012).
Nathan, D.G. & Gunn, R.B. Thalassemia: the consequences of unbalanced hemoglobin synthesis. Am. J. Med. 41, 815–830 (1966).
Schrier, S.L. Pathophysiology of thalassemia. Curr. Opin. Hematol. 9, 123–126 (2002).
Kong, Y. et al. Loss of α-hemoglobin-stabilizing protein impairs erythropoiesis and exacerbates β-thalassemia. J. Clin. Invest. 114, 1457–1466 (2004).
Utsugisawa, T. et al. A road map toward defining the role of Smad signaling in hematopoietic stem cells. Stem Cells 24, 1128–1136 (2006).
Perry, J.M., Harandi, O.F. & Paulson, R.F. BMP4, SCF, and hypoxia cooperatively regulate the expansion of murine stress erythroid progenitors. Blood 109, 4494–4502 (2007).
Socolovsky, M. Molecular insights into stress erythropoiesis. Curr. Opin. Hematol. 14, 215–224 (2007).
De Maria, R. et al. Negative regulation of erythropoiesis by caspase-mediated cleavage of GATA-1. Nature 401, 489–493 (1999).
Bader-Meunier, B. et al. Dyserythropoiesis associated with a Fas-deficient condition in childhood. Br. J. Haematol. 108, 300–304 (2000).
Devadas, S., Zaritskaya, L., Rhee, S.G., Oberley, L. & Williams, M.S. Discrete generation of superoxide and hydrogen peroxide by T cell receptor stimulation: selective regulation of mitogen-activated protein kinase activation and Fas ligand expression. J. Exp. Med. 195, 59–70 (2002).
Dolznig, H. et al. Establishment of normal, terminally differentiating mouse erythroid progenitors: molecular characterization by cDNA arrays. FASEB J. 15, 1442–1444 (2001).
Khandros, E., Thom, C.S., D'Souza, J. & Weiss, M.J. Integrated protein quality-control pathways regulate free α-globin in murine β-thalassemia. Blood 119, 5265–5275 (2012).
Acknowledgements
This work was supported by Agence Nationale de la Recherche (grants ANR-10-JCJC-1108, ANR-12-BSV1-0039, ANR-11-LABX-0051 and ANR-10-BLAN-1109), Assistance Publique Hôpitaux de Paris–CNRS Contrats Hospitaliers de Recherche Translationnelle, Institut National du Cancer, Cancéropôle Île-de-France, Fondation pour la Recherche Médicale, Fondation de France, Association Laurette Fugain and Association pour la Recherche sur le Cancer. The Imagine Institute and the Laboratory of Excellence GR-Ex are funded by the program 'Investissements d'avenir' of the French National Research Agency (ANR-10-IAHU-01 and ANR-11-IDEX-0005-02, respectively). We thank S. Nelson and J. Bex for technical assistance, C. Brouzes for cytological advice, M.G. Traore and N. Goudin for assistance with confocal microscopy and O. Thibaudeau, F. Watier and N. Gadessaud for assistance in histological sample processing. 34-3C IgG2a antibody was provided by S. Izui (University Medical Center, University of Geneva).
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M.D., T.T.M. and A.F. designed and performed all experiments, analyzed the data and helped write the manuscript. J.V., C.C., F.C., D.G., O.N., E. Paubelle and G.C. performed experiments and analyzed data. E. Payen, P.L. and Y.B. provided thalassemic mice, intellectual input and technical expertise for the hemoglobin analysis. J.-A.R. and J.-B.A. provided human samples. T.O.D., R.C. and V.S. participated in project planning, provided RAP-011 and ACE-011, actively contributed to the development of the project and contributed to the writing and editing of the manuscript. Y.Z.G. contributed to the writing and editing of the manuscript. O.H. and I.C.M. supervised the overall project, performed the experiments, analyzed the data and wrote the manuscript.
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T.O.D., R.C. and V.S. are employees of Celgene. This study was partially supported by a grant from Celgene (to O.H.).
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Dussiot, M., Maciel, T., Fricot, A. et al. An activin receptor IIA ligand trap corrects ineffective erythropoiesis in β-thalassemia. Nat Med 20, 398–407 (2014). https://doi.org/10.1038/nm.3468
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DOI: https://doi.org/10.1038/nm.3468
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