Desialylation is a mechanism of Fc-independent platelet clearance and a therapeutic target in immune thrombocytopenia
June Li, Dianne E van der Wal, Guangheng Zhu, Miao Xu, Issaka Yougbare, Li Ma, Brian Vadasz, Naadiya Carrim, Renata Grozovsky, Min Ruan, Lingyan Zhu, Qingshu Zeng, Lili Tao, Zhi-min Zhai, Jun Peng, Ming Hou, Valery Leytin, John Freedman, Karin M Hoffmeister, Heyu Ni, June Li, Dianne E van der Wal, Guangheng Zhu, Miao Xu, Issaka Yougbare, Li Ma, Brian Vadasz, Naadiya Carrim, Renata Grozovsky, Min Ruan, Lingyan Zhu, Qingshu Zeng, Lili Tao, Zhi-min Zhai, Jun Peng, Ming Hou, Valery Leytin, John Freedman, Karin M Hoffmeister, Heyu Ni
Abstract
Immune thrombocytopenia (ITP) is a common bleeding disorder caused primarily by autoantibodies against platelet GPIIbIIIa and/or the GPIb complex. Current theory suggests that antibody-mediated platelet destruction occurs in the spleen, via macrophages through Fc-FcγR interactions. However, we and others have demonstrated that anti-GPIbα (but not GPIIbIIIa)-mediated ITP is often refractory to therapies targeting FcγR pathways. Here, we generate mouse anti-mouse monoclonal antibodies (mAbs) that recognize GPIbα and GPIIbIIIa of different species. Utilizing these unique mAbs and human ITP plasma, we find that anti-GPIbα, but not anti-GPIIbIIIa antibodies, induces Fc-independent platelet activation, sialidase neuraminidase-1 translocation and desialylation. This leads to platelet clearance in the liver via hepatocyte Ashwell-Morell receptors, which is fundamentally different from the classical Fc-FcγR-dependent macrophage phagocytosis. Importantly, sialidase inhibitors ameliorate anti-GPIbα-mediated thrombocytopenia in mice. These findings shed light on Fc-independent cytopenias, designating desialylation as a potential diagnostic biomarker and therapeutic target in the treatment of refractory ITP.
Figures
References
- Rodeghiero F. et al.. Standardization of terminology, definitions and outcome criteria in immune thrombocytopenic purpura of adults and children: report from an international working group. Blood 113, 2386–2393 (2009).
- McMillan R. The pathogenesis of chronic immune thrombocytopenic purpura. Semin. Hematol. 44, S3–S11 (2007).
- Cines D. B., Bussel J. B., Liebman H. A. & Luning Prak E. T. The ITP syndrome: pathogenic and clinical diversity. Blood 113, 6511–6521 (2009).
- Mathias S. D. et al.. Impact of chronic Immune Thrombocytopenic Purpura (ITP) on health-related quality of life: a conceptual model starting with the patient perspective. Health Qual. Life Outcomes 6, 13 (2008).
- Provan D. et al.. International consensus report on the investigation and management of primary immune thrombocytopenia. Blood 115, 168–186 (2010).
- Vianelli N. et al.. Efficacy and safety of splenectomy in immune thrombocytopenic purpura: long-term results of 402 cases. Haematologica 90, 72–77 (2005).
- Neunert C. et al.. The American Society of Hematology 2011 evidence-based practice guideline for immune thrombocytopenia. Blood 117, 4190–4207 (2011).
- Harrington W. J., Minnich V., Hollingsworth J. W. & Moore C. V. Demonstration of a thrombocytopenic factor in the blood of patients with thrombocytopenic purpura. J. Lab. Clin. Med. 38, 1–10 (1951).
- Beardsley D. S. & Ertem M. Platelet autoantibodies in immune thrombocytopenic purpura. Transfus. Sci. 19, 237–244 (1998).
- Bussel J. B. Fc receptor blockade and immune thrombocytopenic purpura. Semin. Hematol. 37, 261–266 (2000).
- Nieswandt B., Bergmeier W., Rackebrandt K., Gessner J. E. & Zirngibl H. Identification of critical antigen-specific mechanisms in the development of immune thrombocytopenic purpura in mice. Blood 96, 2520–2527 (2000).
- Webster M. L. et al.. Relative efficacy of intravenous immunoglobulin G in ameliorating thrombocytopenia induced by antiplatelet GPIIbIIIa versus GPIbalpha antibodies. Blood 108, 943–946 (2006).
- Go R. S., Johnston K. L. & Bruden K. C. The association between platelet autoantibody specificity and response to intravenous immunoglobulin G in the treatment of patients with immune thrombocytopenia. Haematologica 92, 283–284 (2007).
- Peng J. et al.. Association of autoantibody specificity and response to intravenous immunoglobulin G therapy in immune thrombocytopenia: a multicenter cohort study. J. Thromb. Haemost. 12, 497–504 (2014).
- Chang M. et al.. Immune thrombocytopenic purpura (ITP) plasma and purified ITP monoclonal autoantibodies inhibit megakaryocytopoiesis in vitro. Blood 102, 887–895 (2003).
- Zeng Q. et al.. Relative efficacy of steroid therapy in immune thrombocytopenia mediated by anti-platelet GPIIbIIIa versus GPIbalpha antibodies. Am. J. Hematol. 87, 206–208 (2012).
- Ozaki Y., Asazuma N., Suzuki-Inoue K. & Berndt M. C. Platelet GPIb-IX-V-dependent signaling. J. Thromb. Haemost. 3, 1745–1751 (2005).
- Shattil S. J. & Newman P. J. Integrins: dynamic scaffolds for adhesion and signaling in platelets. Blood 104, 1606–1615 (2004).
- Du X. Signaling and regulation of the platelet glycoprotein Ib-IX-V complex. Curr. Opin. Hematol. 14, 262–269 (2007).
- Okumura I., Lombart C. & Jamieson G. A. Platelet glycocalicin. II. Purification and characterization. J. Biol. Chem. 251, 5950–5955 (1976).
- Solum N. O., Hagen I., Filion-Myklebust C. & Stabaek T. Platelet glycocalicin. Its membrane association and solubilization in aqueous media. Biochim. Biophys. Acta 597, 235–246 (1980).
- Rumjantseva V. et al.. Dual roles for hepatic lectin receptors in the clearance of chilled platelets. Nat. Med. 15, 1273–1280 (2009).
- Wandall H. H. et al.. Galactosylation does not prevent the rapid clearance of long-term, 4 degrees C-stored platelets. Blood 111, 3249–3256 (2008).
- Sorensen A. L. et al.. Role of sialic acid for platelet life span: exposure of beta-galactose results in the rapid clearance of platelets from the circulation by asialoglycoprotein receptor-expressing liver macrophages and hepatocytes. Blood 114, 1645–1654 (2009).
- Li C. et al.. The maternal immune response to fetal platelet GPIbalpha causes frequent miscarriage in mice that can be prevented by intravenous IgG and anti-FcRn therapies. J. Clin. Invest. 121, 4537–4547 (2011).
- Lei X. et al.. Anfibatide, a novel GPIb complex antagonist, inhibits platelet adhesion and thrombus formation in vitro and in vivo in murine models of thrombosis. Thromb. Haemost. 111, 279–289 (2014).
- Yang H. et al.. Fibrinogen and von Willebrand factor-independent platelet aggregation in vitro and in vivo. J. Thromb. Haemost. 4, 2230–2237 (2006).
- Reheman A. et al.. Plasma fibronectin depletion enhances platelet aggregation and thrombus formation in mice lacking fibrinogen and von Willebrand factor. Blood 113, 1809–1817 (2009).
- Wang Y. et al.. Plasma fibronectin supports hemostasis and regulates thrombosis. J. Clin. Invest. 124, 4281–4293 (2014).
- Yanabu M. et al.. Tyrosine phosphorylation and p72syk activation by an anti-glycoprotein Ib monoclonal antibody. Blood 89, 1590–1598 (1997).
- Cauwenberghs N. et al.. Fc-receptor dependent platelet aggregation induced by monoclonal antibodies against platelet glycoprotein Ib or von Willebrand factor. Thromb. Haemost. 85, 679–685 (2001).
- Shattil S. J., Hoxie J. A., Cunningham M. & Brass L. F. Changes in the platelet membrane glycoprotein IIb.IIIa complex during platelet activation. J. Biol. Chem. 260, 11107–11114 (1985).
- Bergmeier W. et al.. Flow cytometric detection of activated mouse integrin alphaIIbbeta3 with a novel monoclonal antibody. Cytometry 48, 80–86 (2002).
- King M., McDermott P. & Schreiber A. D. Characterization of the Fc gamma receptor on human platelets. Cell. Immunol. 128, 462–479 (1990).
- Pfueller S. L. & Luscher E. F. The effects of aggregated immunoglobulins on human blood platelets in relation to their complement-fixing abilities. II. Structural requirements of the immunoglobulin. J. Immunol. 109, 526–533 (1972).
- Hoffmeister K. M. et al.. Glycosylation restores survival of chilled blood platelets. Science 301, 1531–1534 (2003).
- Zhang Y. et al.. Identification of selective inhibitors for human neuraminidase isoenzymes using C4,C7-modified 2-deoxy-2,3-didehydro-N-acetylneuraminic acid (DANA) analogues. J. Med. Chem. 56, 2948–2958 (2013).
- Judson P. A., Anstee D. J. & Clamp J. R. Isolation and characterization of the major oligosaccharide of human platelet membrane glycoprotein GPIb. Biochem. J. 205, 81–90 (1982).
- Kinlough-Rathbone R. L., Perry D. W., Rand M. L. & Packham M. A. Responses to aggregating agents after cleavage of GPIb of human platelets by the O-sialoglycoprotein endoprotease from Pasteurella haemolytica- potential surrogates for Bernard-Soulier platelets? Thromb. Res. 99, 165–172 (2000).
- Jansen A. J. et al.. Desialylation accelerates platelet clearance after refrigeration and initiates GPIbalpha metalloproteinase-mediated cleavage in mice. Blood 119, 1263–1273 (2012).
- van der Wal D. E. et al.. Role of glycoprotein Ibalpha mobility in platelet function. Thromb. Haemost. 103, 1033–1043 (2010).
- Gitz E. et al.. Improved platelet survival after cold storage by prevention of glycoprotein Ibalpha clustering in lipid rafts. Haematologica 97, 1873–1881 (2012).
- van der Wal D. E. et al.. Platelet apoptosis by cold-induced glycoprotein Ibalpha clustering. J. Thromb. Haemost. 8, 2554–2562 (2010).
- Kasirer-Friede A. et al.. Lateral clustering of platelet GP Ib-IX complexes leads to up-regulation of the adhesive function of integrin alpha IIbbeta 3. J. Biol. Chem. 277, 11949–11956 (2002).
- Marshall J. S. et al.. Measurement of circulating desialylated glycoproteins and correlation with hepatocellular damage. J. Clin. Invest. 54, 555–562 (1974).
- Grozovsky R. et al.. The Ashwell-Morell receptor regulates hepatic thrombopoietin production via JAK2-STAT3 signaling. Nat. Med. 21, 47–54 (2015).
- Alimardani G., Guichard J., Fichelson S. & Cramer E. M. Pathogenic effects of anti-glycoprotein Ib antibodies on megakaryocytes and platelets. Thromb. Haemost. 88, 1039–1046 (2002).
- Cadroy Y. et al.. Relative antithrombotic effects of monoclonal antibodies targeting different platelet glycoprotein-adhesive molecule interactions in nonhuman primates. Blood 83, 3218–3224 (1994).
- Cauwenberghs N. et al.. Antithrombotic effect of platelet glycoprotein Ib-blocking monoclonal antibody Fab fragments in nonhuman primates. Arterioscler. Thromb. Vasc. Biol. 20, 1347–1353 (2000).
- Rubinstein E., Kouns W. C., Jennings L. K., Boucheix C. & Carroll R. C. Interaction of two GPIIb/IIIa monoclonal antibodies with platelet Fc receptor (Fc gamma RII). Br. J. Haematol. 78, 80–86 (1991).
- Sullam P. M. et al.. Physical proximity and functional interplay of the glycoprotein Ib-IX-V complex and the Fc receptor FcgammaRIIA on the platelet plasma membrane. J. Biol. Chem. 273, 5331–5336 (1998).
- Shrimpton C. N. et al.. Localization of the adhesion receptor glycoprotein Ib-IX-V complex to lipid rafts is required for platelet adhesion and activation. J. Exp. Med. 196, 1057–1066 (2002).
- Urbanus R. T. et al.. Patient autoantibodies induce platelet destruction signals via raft-associated glycoprotein Ibalpha and FcgammaRIIa in immune thrombocytopenia. Haematologica 98, e70–e72 (2013).
- Sarpatwari A. et al.. Autologous 111 In-labelled platelet sequestration studies in patients with primary immune thrombocytopenia (ITP) prior to splenectomy: a report from the United Kingdom ITP Registry. Br. J. Haematol. 151, 477–487 (2010).
- Ghanima W., Godeau B., Cines D. B. & Bussel J. B. How I treat immune thrombocytopenia: the choice between splenectomy or a medical therapy as a second-line treatment. Blood 120, 960–969 (2012).
- Enger C. et al.. Hepatobiliary laboratory abnormalities among patients with chronic or persistent immune thrombocytopenia (ITP). Ann. Hepatol. 10, 188–195 (2011).
- Davidson C. J., Tuddenham E. G. & McVey J. H. 450 million years of hemostasis. J. Thromb. Haemost. 1, 1487–1494 (2003).
- Li J. et al.. Severe platelet desialylation in a patient with glycoprotein Ib/IX antibody-mediated immune thrombocytopenia and fatal pulmonary hemorrhage. Haematologica 99, e61–e63 (2014).
- Jansen A. J., Peng J., Zhao H. G., Hou M. & Ni H. Sialidase inhibition to increase platelet counts: a new treatment option for thrombocytopenia. Am. J. Hematol. 90, E94–E95 (2015).
- Shao L. et al.. Successful treatment with oseltamivir phosphate in a patient with chronic immune thrombocytopenia positive for anti-GPIb/IX autoantibody. Platelets 26, 495–497 (2014).
- Nan X., Carubelli I. & Stamatos N. M. Sialidase expression in activated human T lymphocytes influences production of IFN-gamma. J. Leukoc. Biol. 81, 284–296 (2007).
- Chen X. P., Enioutina E. Y. & Daynes R. A. The control of IL-4 gene expression in activated murine T lymphocytes: a novel role for neu-1 sialidase. J. Immunol. 158, 3070–3080 (1997).
- Seyrantepe V. et al.. Regulation of phagocytosis in macrophages by neuraminidase 1. J. Biol. Chem. 285, 206–215 (2010).
- Grewal P. K. et al.. The Ashwell receptor mitigates the lethal coagulopathy of sepsis. Nature medicine 14, 648–655 (2008).
- Tribulatti M. V., Mucci J., Van Rooijen N., Leguizamon M. S. & Campetella O. The trans-sialidase from Trypanosoma cruzi induces thrombocytopenia during acute Chagas' disease by reducing the platelet sialic acid contents. Infect. Immun. 73, 201–207 (2005).
- Alioglu B., Tasar A., Ozen C., Selver B. & Dallar Y. An experience of oseltamivir phosphate (tamiflu) in a pediatric patient with chronic idiopathic thrombocytopenic purpura: a case report. Pathophysiol. Haemost. Thromb. 37, 55–58 (2010).
- Mandic R., Opper C., Krappe J. & Wesemann W. Platelet sialic acid as a potential pathogenic factor in coronary heart disease. Thromb. Res. 106, 137–141 (2002).
- Butun I. I. et al.. Preliminary study showing the relationship between platelet fibronectin, sialic acid, and ADP-induced aggregation levels in coronary heart disease. Clin. Appl. Thromb. Hemost. 13, 308–312 (2007).
- McGill D. A. & Ardlie N. G. Abnormal platelet reactivity in men with premature coronary heart disease. Coron. Artery Dis. 5, 889–900 (1994).
- Wang Y. et al.. Tyrosine phosphatase MEG2 modulates murine development and platelet and lymphocyte activation through secretory vesicle function. J. Exp. Med. 202, 1587–1597 (2005).
Source: PubMed