Mathematical Model of Platelet Intracellular Signaling After Activation by Fucoidan
Blood platelets are the cells responsible for prevention of the blood loss. Fucoidan is a brown algae extract that is known to activate platelets via C-type lectin receptor of the second type. On the other hand, different fucoidans are now considered as perspective immunomodulators. Thus, application of fucoidan as a medicinal drug seems to be contradictory. In this work we studied activation of platelets by fucoidan in silico and in vitro. The computational model describes the behavior of the participants of the fucoidan receptor signaling cascade. The model was validated with available experimental data published earlier. In order to confirm the model predictions, the fucoidan-induced activation of platelets was assessed in flow cytometry and aggregometry experiments. The resultant model describes changes in the activity of tyrosine kinases of Syk and Sarc family and subsequent activation of phospholipase Cγ2. One of the main model prediction is a significant increase in the platelet cytosolic calcium level after the activation by fucoidan. This prediction was confirmed in the experiments. Thus, fucoidan, as a true platelet activator, cannot be applied in therapy.
Keywords:intracellular signaling computer modeling flow cytometry
Authors are grateful to Prof. F.I. Ataullakhanov for valuable discussions and advise during the research process, as well as to Dr. N.E. Ustyuzhanina for discussions concerning fucoidan.
The work was supported by the Russian Foundation for Basic Research (project no. 17-54-04009), Belorussian Foundation for Basic Research (project no. B17RM-006), and by the President’s grants for young investigators (project nos. MK-5879.2016.4 and MD-229.2017.4).
COMPLIANCE WITH ETHICAL STANDARDS
Conflict of interests. The authors declare that they have no conflict of interest.
Statement of compliance with standards of research involving humans as subjects. All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards. Informed consent was obtained from all individual participants involved in the study.
- 1.Cumashi A., Ushakova N.A., Preobrazhenskaya M.E., D’Incecco A., Piccoli A., Totani L., Tinari N., Morozevich G.E., Berman A.E., Bilan M.I., Usov A.I., Ustyuzhanina N.E., Grachev A.A., Sanderson C.J., Kelly M., Rabinovich G.A., Iacobelli S., Nifantiev N.E. 2007. A comparative study of the anti-inflammatory, anticoagulant, antiangiogenic, and antiadhesive activities of nine different fucoidans from brown seaweeds. Glycobiology. 17 (5), 541–552.CrossRefGoogle Scholar
- 4.Shirai T., Inoue O., Tamura S., Tsukiji N., Sasaki T., Endo H., Satoh K., Osada M., Sato-Uchida H., Fujii H., Ozaki Y., Suzuki-Inoue K. 2017. C-type lectin-like receptor 2 promotes hematogenous tumor metastasis and prothrombotic state in tumor-bearing mice. J. Thromb. Haemost. 15 (3), 513–525.CrossRefGoogle Scholar
- 6.Suzuki-Inoue K., Inoue O., Ding G., Nishimura S., Hokamura K., Eto K., Kashiwagi H., Tomiyama Y., Yatomi Y., Umemura K., Shin Y., Hirashima M., Ozaki Y. 2010. Essential in vivo roles of the C-type lectin receptor CLEC-2: Embryonic/neonatal lethality of CLEC-2-deficient mice by blood/lymphatic misconnections and impaired thrombus formation of CLEC-2-deficient platelets. J. Biol. Chem. 285 (32), 24494–24507.CrossRefGoogle Scholar
- 7.Sveshnikova A.N., Balatskiy A.V., Demianova A.S., Shepelyuk T.O., Shakhidzhanov S.S., Balatskaya M.N., Pichugin A.V., Ataullakhanov F.I., Panteleev M.A. 2016. Systems biology insights into the meaning of the platelet’s dual-receptor thrombin signaling. J. Thromb. Haemost. 14 (10), 2045–2057.CrossRefGoogle Scholar
- 8.Gibbins J.M., Mahaut-Smith M.P. 2004. Platelets and megakaryocytes. Vol. 1. Functional assays. New Jersy: Humana Press, Inc.Google Scholar
- 14.Pollitt A.Y., Poulter N.S., Gitz E., Navarro-Nuñez L., Wang Y.J., Hughes C.E., Thomas S.G., Nieswandt B., Douglas M.R., Owen D.M., Jackson D.G., Dustin M.L., Watson S.P. 2014. Syk and src family kinases regulate c-type lectin receptor 2 (clec-2)-mediated clustering of podoplanin and platelet adhesion to lymphatic endothelial cells. J. Biol. Chem. 289 (52), 35695–35710.CrossRefGoogle Scholar
- 15.Hughes C.E., Sinha U., Pandey A., Eble J.A., O’Callaghan C.A., Watson S.P. 2013. Critical role for an acidic amino acid region in platelet signaling by the HemITAM (hemi-immunoreceptor tyrosine-based activation motif) containing receptor CLEC-2 (C-type lectin receptor-2). J. Biol. Chem. 288 (7), 5127–5135.CrossRefGoogle Scholar
- 20.Pasquet J.M., Gross B., Quek L., Asazuma N., Zhang W., Sommers C.L., Schweighoffer E., Tybulewicz V., Judd B., Lee J.R., Koretzky G., Love P.E., Samelson L.E., Watson S.P. 1999. LAT is required for tyrosine phosphorylation of phospholipase cgamma2 and platelet activation by the collagen receptor GPVI. Mol. Cell. Biol. 19 (12), 8326–8334.CrossRefGoogle Scholar
- 21.Gibbins J.M., Briddon S., Shutes A., Van Vugt M.J., Van De Winkel J.G.J., Saito T., Watson S.P. 1998. The p85 subunit of phosphatidylinositol 3-kinase associates with the Fc receptor γ-chain and linker for activitor of T cells (LAT) in platelets stimulated by collagen and convulxin. J. Biol. Chem. 273 (51), 34437–34443.CrossRefGoogle Scholar
- 26.Musumeci L., Kuijpers M.J., Gilio K., Hego A., Theatre E., Maurissen L., Vandereyken M., Diogo C.V., Lecut C., Guilmain W., Bobkova E.V., Eble J.A., Dahl R., Drion P., Rascon J., Mostofi Y., Yuan H., Sergienko E., Chung T.D.Y., Thiry M., Senis Y., Moutschen M., Mustelin T., Lancellotti P., Heemskerk J.W.M., Tautz L., Oury C., Rahmouni S. 2015. Dual-specificity phosphatase 3 deficiency or inhibition limits platelet activation and arterial thrombosis. Circulation. 131 (7), 656–668.CrossRefGoogle Scholar
- 31.Sklar L.A., Jesaitis A.J., Painter R.G., Cochrane C.G. 1981. The kinetics of neutrophil activation. The response to chemotactic peptides depends upon whether ligand–receptor interaction is rate-limiting. J. Biol. Chem. 256 (19), 9909–9914.Google Scholar
- 32.Michelson A.D. 2013. Platelets. London: Elsevier.Google Scholar
- 39.Burkhart J.M., Vaudel M., Gambaryan S., Radau S., Walter U., Martens L., Geiger J., Sickmann A., Zahedi R.P. 2012. The first comprehensive and quantitative analysis of human platelet protein composition allows the comparative analysis of structural and functional pathways. Blood. 120 (15), e73–e82.CrossRefGoogle Scholar