Subscribe to RSS
DOI: 10.1055/a-2398-9532
Shear Stress Promotes Remodeling of Platelet Glycosylation via Upregulation of Platelet Glycosidase Activity: One More Thing
Funding This research is supported by grants from the American Heart Association (Career Development Award 935890 to Yana Roka-Moiia), the University of Arizona Sarver Heart Center (Jack and Mildred Michelson Cardiovascular Research Award and John H. Midkiff Cardiovascular Research Award to Yana Roka-Moiia), and the Arizona Center for Accelerated Biomedical Innovation (ACABI) of the University of Arizona (an unrestricted grant to Marvin J. Slepian).
Abstract
Background Mechanical circulatory support (MCS) is a mainstay of therapy for advanced and end-stage heart failure. Accompanied by systemic anticoagulation, contemporary MCS has become less thrombogenic, with bleeding complications emerging as a major cause of readmission and 1-year mortality. Shear-mediated platelet dysfunction and thrombocytopenia of undefined etiology are primary drivers of MCS-related bleeding. Recently, it has been demonstrated that deprivation of platelet surface glycosylation is associated with the decline of hemostatic function, microvesiculation, and premature apoptosis. We test the hypothesis that shear stress induces remodeling of platelet surface glycosylation via upregulation of glycosidase activity, thus facilitating platelet count decline and intense microvesiculation.
Methods Human gel-filtered platelets were exposed to continuous shear stress in vitro. Platelets and platelet-derived microparticles (PDMPs) were quantified via flow cytometry using size standard fluorescent nanobeads. Platelet surface glycosylation and NEU1 expression were evaluated using lectin- or immune-staining and multicolor flow cytometry; lectin blotting was utilized to verify glycosylation of individual glycoproteins. Platelet neuraminidase, galactosidase, hexosaminidase, and mannosidase activities were quantified using 4-methylumbelliferone-based fluorogenic substrates.
Results We demonstrate that shear stress promotes selective remodeling of platelet glycosylation via downregulation of 2,6-sialylation, terminal galactose, and mannose, while 2,3-sialylation remains largely unchanged. Shear-mediated deglycosylation is partially attenuated by neuraminidase inhibitors, strongly suggesting the involvement of platelet neuraminidase in observed phenomena. Shear stress increases platelet NEU1 surface expression and potentiates generation of numerous NEU1+ PDMPs. Platelets exhibit high basal hexosaminidase and mannosidase activities; basal activities of platelet neuraminidase and galactosidase are rather low and are significantly upregulated by shear stress. Shear stress of increased magnitude and duration promotes an incremental decline of platelet count and immense microvesiculation, both being further exacerbated by neuraminidase and partially attenuated by neuraminidase inhibition.
Conclusion Our data indicate that shear stress accumulation, consistent with supraphysiologic conditions of device-supported circulation, promotes remodeling of platelet glycosylation via selective upregulation of platelet glycosidase activity. Shear-mediated platelet deglycosylation is associated with platelet count drop and increased microvesiculation, thus offering a direct link between deglycosylation and thrombocytopenia observed in device-supported patients. Based on our findings, we propose a panel of molecular markers to be used for reliable detection of shear-mediated platelet deglycosylation in MCS.
Keywords
platelet deglycosylation - shear stress - neuraminidase - mechanical circulatory support - thrombocytopeniaAuthors' Contribution
Y. Roka-Moiia designed the study, performed experiments, analyzed and interpreted data, wrote and edited the article, and acquired funding; S. Lewis and E. Cleveland performed experiments; J. E. Italiano participated in discussions and edited the article; M. J. Slepian participated in discussions, edited the article, and acquired funding.
Publication History
Received: 27 March 2024
Accepted: 20 August 2024
Accepted Manuscript online:
21 August 2024
Article published online:
12 September 2024
© 2024. Thieme. All rights reserved.
Georg Thieme Verlag KG
Stuttgart · New York
-
References
- 1 Starling RC, Moazami N, Silvestry SC. et al. Unexpected abrupt increase in left ventricular assist device thrombosis. N Engl J Med 2014; 370 (01) 33-40
- 2 Koliopoulou A, McKellar SH, Rondina M, Selzman CH. Bleeding and thrombosis in chronic ventricular assist device therapy: focus on platelets. Curr Opin Cardiol 2016; 31 (03) 299-307
- 3 Goldstein DJ, John R, Salerno C. et al. Algorithm for the diagnosis and management of suspected pump thrombus. J Heart Lung Transplant 2013; 32 (07) 667-670
- 4 Rogers JG. Managing VAD complications. J Am Coll Cardiol 2016; 67 (23) 2769-2771
- 5 Subramaniam AV, Barsness GW, Vallabhajosyula S, Vallabhajosyula S. Complications of temporary percutaneous mechanical circulatory support for cardiogenic shock: an appraisal of contemporary literature. Cardiol Ther 2019; 8 (02) 211-228
- 6 Lakshmanan S, Cuker A. Thrombocytopenia in the intensive care unit and after solid organ transplantation. In: The Coagulation Consult. New York, NY: Springer; 2014: 115-132
- 7 Houry EA, Gengler BE, Alberts JL, Van Tuyl JS. Evaluation of thrombocytopenia in patients receiving percutaneous mechanical circulatory support with an Impella device. Crit Care Explor 2022; 4 (10) e0772
- 8 Dwaah H, Jain N, Kapur NK. et al. The impact of temporary mechanical circulatory support strategies on thrombocytopenia. J Crit Care 2023; 73: 154216
- 9 Wang L, Shao J, Shao C, Wang H, Jia M, Hou X. The relative early decrease in platelet count is associated with mortality in post-cardiotomy patients undergoing venoarterial extracorporeal membrane oxygenation. Front Med (Lausanne) 2021; 8 (November): 733946
- 10 Koupenova M, Clancy L, Corkrey HA, Freedman JE. Circulating platelets as mediators of immunity, inflammation, and thrombosis. Circ Res 2018; 122 (02) 337-351
- 11 Slepian MJ, Sheriff J, Hutchinson M. et al. Shear-mediated platelet activation in the free flow: perspectives on the emerging spectrum of cell mechanobiological mechanisms mediating cardiovascular implant thrombosis. J Biomech 2017; 50: 20-25
- 12 Roka-Moiia Y, Walk R, Palomares DE. et al. Platelet activation via shear stress exposure induces a differing pattern of biomarkers of activation versus biochemical agonists. Thromb Haemost 2020; 120 (05) 776-792
- 13 Roka-Moiia Y, Miller-Gutierrez S, Palomares DE. et al. Platelet dysfunction during mechanical circulatory support: elevated shear stress promotes downregulation of αIIbβ3 and GPIb via microparticle shedding decreasing platelet aggregability. Arterioscler Thromb Vasc Biol 2021; 41 (04) 1319-1336
- 14 Leytin V, Allen DJ, Mykhaylov S. et al. Pathologic high shear stress induces apoptosis events in human platelets. Biochem Biophys Res Commun 2004; 320 (02) 303-310
- 15 Roka-Moiia Y, Ammann KR, Miller-Gutierrez S. et al. Shear-mediated platelet microparticles demonstrate phenotypic heterogeneity as to morphology, receptor distribution, and hemostatic function. Int J Mol Sci 2023; 24 (08) 7386
- 16 Roka-Moiia Y, Li M, Ivich A, Muslmani S, Kern KB, Slepian MJ. Impella 5.5 versus Centrimag: a head-to-head comparison of device hemocompatibility. ASAIO J 2020; 66 (10) 1142-1151
- 17 Quach ME, Chen W, Li R. Mechanisms of platelet clearance and translation to improve platelet storage. Blood 2018; 131 (14) 1512-1521
- 18 Cai X, Thinn AMM, Wang Z, Shan H, Zhu J. The importance of N-glycosylation on β3 integrin ligand binding and conformational regulation. Sci Rep 2017; 7 (01) 4656
- 19 Lee MM, Nasirikenari M, Manhardt CT. et al. Platelets support extracellular sialylation by supplying the sugar donor substrate. J Biol Chem 2014; 289 (13) 8742-8748
- 20 Lee-Sundlov MM, Ashline DJ, Hanneman AJ. et al. Circulating blood and platelets supply glycosyltransferases that enable extrinsic extracellular glycosylation. Glycobiology 2017; 27 (02) 188-198
- 21 Li R, Hoffmeister KM, Falet H. Glycans and the platelet life cycle. Platelets 2016; 27 (06) 505-511
- 22 Hollenhorst MA, Tiemeyer KH, Mahoney KE. et al. Comprehensive analysis of platelet glycoprotein Ibα ectodomain glycosylation. J Thromb Haemost 2023; 21 (04) 995-1009
- 23 Andrews RK, Gardiner EE. Metalloproteolytic receptor shedding…platelets “acting their age”. Platelets 2016; 27 (06) 512-518
- 24 Li Y, Fu J, Ling Y. et al. Sialylation on O-glycans protects platelets from clearance by liver Kupffer cells. Proc Natl Acad Sci U S A 2017; 114 (31) 8360-8365
- 25 Giannini S, Falet H, Hoffmeister K. Platelet glycobiology and the control of platelet function and lifespan. In: Platelets 2019; 79-97
- 26 Tiemeyer KH, Kuter DJ, Cairo CW, Hollenhorst MA. New insights into the glycobiology of immune thrombocytopenia. Curr Opin Hematol 2023; 30 (06) 210-218
- 27 Pikoula M, Tessier MB, Woods RJ, Ventikos Y. Oligosaccharide model of the vascular endothelial glycocalyx in physiological flow. Microfluid Nanofluidics 2018; 22 (02) 21
- 28 Nobili M, Sheriff J, Morbiducci U, Redaelli A, Bluestein D. Platelet activation due to hemodynamic shear stresses: damage accumulation model and comparison to in vitro measurements. ASAIO J 2008; 54 (01) 64-72
- 29 Sheriff J, Soares JS, Xenos M, Jesty J, Slepian MJ, Bluestein D. Evaluation of shear-induced platelet activation models under constant and dynamic shear stress loading conditions relevant to devices. Ann Biomed Eng 2013; 41 (06) 1279-1296
- 30 Sheriff J, Tran PL, Hutchinson M. et al. Repetitive hypershear activates and sensitizes platelets in a dose-dependent manner. Artif Organs 2016; 40 (06) 586-595
- 31 Bojar D, Meche L, Meng G. et al. A useful guide to lectin binding: machine-learning directed annotation of 57 unique lectin specificities. ACS Chem Biol 2022; 17 (11) 2993-3012
- 32 Schmitz G, Rothe G, Ruf A. et al. European Working Group on Clinical Cell Analysis: consensus protocol for the flow cytometric characterisation of platelet function. Thromb Haemost 1998; 79 (05) 885-896
- 33 Josefsson EC, Ramström S, Thaler J, Lordkipanidzé M. COAGAPO study group. Consensus report on markers to distinguish procoagulant platelets from apoptotic platelets: communication from the Scientific and Standardization Committee of the ISTH. J Thromb Haemost 2023; 21 (08) 2291-2299
- 34 Ramírez-López A, Álvarez Román MT, Monzón Manzano E. et al. The importance of platelet glycoside residues in the haemostasis of patients with immune thrombocytopaenia. J Clin Med 2021; 10 (08) 1661
- 35 Lombardo A, Caimi L, Marchesini S, Goi GC, Tettamanti G. Enzymes of lysosomal origin in human plasma and serum: assay conditions and parameters influencing the assay. Clin Chim Acta 1980; 108 (03) 337-346
- 36 Li J, van der Wal DE, Zhu G. et al. Desialylation is a mechanism of Fc-independent platelet clearance and a therapeutic target in immune thrombocytopenia. Nat Commun 2015; 6 (01) 7737
- 37 Jansen AJG, Josefsson EC, Rumjantseva V. et al. Desialylation accelerates platelet clearance after refrigeration and initiates GPIbα metalloproteinase-mediated cleavage in mice. Blood 2012; 119 (05) 1263-1273
- 38 van der Wal DE, Davis AM, Mach M, Marks DC. The role of neuraminidase 1 and 2 in glycoprotein Ibα-mediated integrin αIIbβ3 activation. Haematologica 2020; 105 (04) 1081-1094
- 39 Burkhart JM, Vaudel M, Gambaryan S. et al. The first comprehensive and quantitative analysis of human platelet protein composition allows the comparative analysis of structural and functional pathways. Blood 2012; 120 (15) e73-e82
- 40 Kullaya V, de Jonge MI, Langereis JD. et al. Desialylation of platelets by pneumococcal neuraminidase a induces ADP-dependent platelet hyperreactivity. Infect Immun 2018; 86 (10) e00213-18
- 41 Li MF, Li XL, Fan KL. et al. Platelet desialylation is a novel mechanism and a therapeutic target in thrombocytopenia during sepsis: an open-label, multicenter, randomized controlled trial. J Hematol Oncol 2017; 10 (01) 104
- 42 Nigam PK, Narain VS, Kumar A, Nigam PK. Sialic acid in cardiovascular diseases. Indian J Clin Biochem 2006; 21 (01) 54-61
- 43 Keil JM, Rafn GR, Turan IM, Aljohani MA, Sahebjam-Atabaki R, Sun XL. Sialidase inhibitors with different mechanisms. J Med Chem 2022; 65 (20) 13574-13593
- 44 Almahayni K, Spiekermann M, Fiore A, Yu G, Pedram K, Möckl L. Small molecule inhibitors of mammalian glycosylation. Matrix Biol Plus 2022; 16: 100108
- 45 Shi D, Yang J, Yang D. et al. Anti-influenza prodrug oseltamivir is activated by carboxylesterase human carboxylesterase 1, and the activation is inhibited by antiplatelet agent clopidogrel. J Pharmacol Exp Ther 2006; 319 (03) 1477-1484
- 46 Rudakova EV, Boltneva NP, Makhaeva GF. Comparative analysis of esterase activities of human, mouse, and rat blood. Bull Exp Biol Med 2011; 152 (01) 73-75
- 47 Goth CK, Halim A, Khetarpal SA, Rader DJ, Clausen H, Schjoldager KT-BG. A systematic study of modulation of ADAM-mediated ectodomain shedding by site-specific O-glycosylation. Proc Natl Acad Sci U S A 2015; 112 (47) 14623-14628
- 48 Karabasheva D, Cole NB, Donaldson JG. Roles for trafficking and O-linked glycosylation in the turnover of model cell surface proteins. J Biol Chem 2014; 289 (28) 19477-19490
- 49 Wang Y, Chen W, Zhang W. et al. Desialylation of O-glycans on glycoprotein Ibα drives receptor signaling and platelet clearance. Haematologica 2020; 105 (05) 220
- 50 Sumida M, Hane M, Yabe U. et al. Rapid trimming of cell surface polysialic acid (PolySia) by exovesicular sialidase triggers release of preexisting surface neurotrophin. Chem 2015; 290 (21) 13202-13214
- 51 Delaveris CS, Wang CL, Riley NM, Li S, Kulkarni RU, Bertozzi CR. Microglia mediate contact-independent neuronal network remodeling via secreted neuraminidase-3 associated with extracellular vesicles. ACS Cent Sci 2023; 9 (11) 2108-2114
- 52 Holmsen H, Setkowsky CA, Lages B, Day HJ, Weiss HJ, Scrutton MC. Content and thrombin-induced release of acid hydrolases in gel-filtered platelets from patients with storage pool disease. Blood 1975; 46 (01) 131-142
- 53 Holmsen H, Weiss HJ. Secretable storage pools in platelets. Annu Rev Med 1979; 30 (01) 119-134
- 54 Haslund-Gourley BS, Aziz PV, Heithoff DM. et al. Establishment of blood glycosidase activities and their excursions in sepsis. PNAS Nexus 2022; 1 (03) pgac113
- 55 Ostrowska H, Krukowska K, Kalinowska J, Orłowska M, Lengiewicz I. Lysosomal high molecular weight multienzyme complex. Cell Mol Biol Lett 2003; 8 (01) 19-24
- 56 Maurice P, Baud S, Bocharova OV. et al. New insights into molecular organization of human neuraminidase-1: transmembrane topology and dimerization ability. Sci Rep 2016; 6 (November): 38363
- 57 Gorelik A, Illes K, Mazhab-Jafari MT, Nagar B. Structure of the immunoregulatory sialidase NEU1. Sci Adv 2023; 9 (20) eadf8169
- 58 Hata K, Koseki K, Yamaguchi K. et al. Limited inhibitory effects of oseltamivir and zanamivir on human sialidases. Antimicrob Agents Chemother 2008; 52 (10) 3484-3491
- 59 Glanz VY, Myasoedova VA, Grechko AV, Orekhov AN. Inhibition of sialidase activity as a therapeutic approach. Drug Des Devel Ther 2018; 12: 3431-3437
- 60 Guo Z, Fan D, Liu FY. et al. NEU1 regulates mitochondrial energy metabolism and oxidative stress post-myocardial infarction in mice via the SIRT1/PGC-1 alpha axis. Front Cardiovasc Med 2022; 9 (April): 821317
- 61 Heimerl M, Sieve I, Ricke-Hoch M. et al. Neuraminidase-1 promotes heart failure after ischemia/reperfusion injury by affecting cardiomyocytes and invading monocytes/macrophages. Basic Res Cardiol 2020; 115 (06) 62
- 62 Rauch A, Dupont A, Rosa M. et al. Shear Forces Induced Platelet Clearance Is a New Mechanism of Thrombocytopenia. Circ Res 2023; 133 (10) 826-841
- 63 van der Pol E, Hoekstra AG, Sturk A, Otto C, van Leeuwen TG, Nieuwland R. Optical and non-optical methods for detection and characterization of microparticles and exosomes. J Thromb Haemost 2010; 8 (12) 2596-2607
- 64 Brinkman-Van der Linden ECM, Sonnenburg JL, Varki A. Effects of sialic acid substitutions on recognition by Sambucus nigra agglutinin and Maackia amurensis hemagglutinin. Anal Biochem 2002; 303 (01) 98-104