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  • Basic Science Article
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Comparison of platelet proteomic profiles between children and adults reveals origins of functional differences

Pediatric Research volume 95, pages 966–973 (2024)Cite this article

Abstract

Background

Platelets are blood cells responsible for the prevention of blood loss upon vessel wall disruption. It has been demonstrated that platelet functioning differs significantly between adult and pediatric donors. This study aimed to identify potential differences between the protein composition of platelets of pediatric, adolescent, and adult donors.

Methods

Platelet functional testing was conducted with live cell flow cytometry. Using a straightforward approach to platelet washing based on the sequential platelets centrifugation-resuspension, we were able to obtain stable and robust proteomics results, which corresponded to previously published data.

Results

We have identified that pediatric donors’ platelets have increased amounts of proteins, responsible for mitochondrial activity, proteasome activity, and vesicle transport. Flow cytometry analysis of platelet intracellular signaling and functional responses revealed that platelets of the pediatric donors have diminished granule secretion and increased quiescent platelet calcium concentration and decreased calcium mobilization in response to ADP. We could explain the observed changes in calcium responses by the increased mitochondria protein content, and the changes in granule secretion could be explained by the differences in vesicle transport protein content.

Conclusions

Therefore, we can conclude that the age-dependence of platelet functional responses originates from the difference in platelet protein content.

Impact

  • Platelets of infants are known to functionally differ from the platelet of adult donors, although the longevity and persistivity of these differences are debatable.

  • Pediatric donor platelets have enhanced amounts of mitochondrial, proteasomal, and vesicle transport proteins.

  • Platelets of the pediatric donors had increased cytosolic calcium in the resting state, what is explained by the increased numbers of mitochondrial proteins. Infants had decreased platelet granule release, which resolved upon adolescence. Thus, platelets of the infants should be assessed differently from adult platelets.

  • Differences in platelet proteomic contents persisted in adolescent groups, yet, no significant differences in platelet function were observed.

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Fig. 1: Global proteomic analysis of platelets.
Fig. 2: Comparison between children, adolescent and adult platelet proteomes.
Fig. 3: Clustering analysis between differentially presented proteins.
Fig. 4: Analysis of the platelet intracellular signaling.
Fig. 5: End-point flow cytometry.

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Data availability

The datasets generated during and/or analyzed during the current study are available from the corresponding author upon reasonable request.

References

  1. Sveshnikova, A. et al. Platelet functional responses and signalling: the molecular relationship. Part 1: responses. Syst. Biol. Physiol. Rep. 1, 20 (2021).

    Article  Google Scholar 

  2. Panteleev, M. A. & Sveshnikova, A. N. Platelets and hemostasis. Oncohematology 9, 65–73 (2014).

    Google Scholar 

  3. Cuenca-Zamora, E. J., Ferrer-Marín, F., Rivera, J., Teruel-Montoya, R. Tubulin in platelets: when the shape matters. Int. J. Mol. Sci. 20. https://doi.org/10.3390/ijms20143484 (2019).

  4. Gao, J. et al. Kindlin supports platelet integrin αIIbβ3 activation by interacting with paxillin. J. Cell Sci. 130, 3764–3775 (2017).

    CAS  PubMed  PubMed Central  Google Scholar 

  5. Deppermann, C. et al. Platelet secretion is crucial to prevent bleeding in the ischemic brain but not in the inflamed skin or lung in mice. Blood 129, 1702–1706 (2017).

    Article  CAS  PubMed  Google Scholar 

  6. Podoplelova, N. A., Nechipurenko, D. Y., Ignatova, A. A., Sveshnikova, A. N. & Panteleev, M. A. Procoagulant platelets: mechanisms of generation and action. Hämostaseologie 41, 146–153 (2021).

    Article  CAS  PubMed  Google Scholar 

  7. Dale, G. L. Coated-platelets: an emerging component of the procoagulant response. J. Thromb. Haemost. 3, 2185–2192 (2005).

    Article  CAS  PubMed  Google Scholar 

  8. Hézard, N. et al. Unexpected persistence of platelet hyporeactivity beyond the neonatal period: a flow cytometric study in neonates, infants and older children. Thromb. Haemost. 90, 116–123 (2003).

    Article  PubMed  Google Scholar 

  9. Herken, K. et al. Age-dependent control of collagen-dependent platelet responses by thrombospondin-1-comparative analysis of platelets from neonates, children, adolescents, and adults. Int J. Mol. Sci. 22, 4883 (2021).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Yip, C., Linden, M. D., Attard, C., Monagle, P. & Ignjatovic, V. Platelets from children are hyper-responsive to activation by thrombin receptor activator peptide and adenosine diphosphate compared to platelets from adults. Br. J. Haematol. 168, 526–532 (2015).

    Article  CAS  PubMed  Google Scholar 

  11. Ignatova, A. A. et al. Flow cytometry for pediatric platelets. Platelets 30, 428–437 (2019).

    Article  CAS  PubMed  Google Scholar 

  12. Blair, P. & Flaumenhaft, R. Platelet α–granules: basic biology and clinical correlates. Blood Rev. 23, 177–189 (2009).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. McNicol, A. & Israels, S. J. Platelet dense granules: structure, function and implications for haemostasis. Thromb. Res. 95, 1–18 (1999).

    Article  CAS  PubMed  Google Scholar 

  14. Tesfamariam, B. Distinct characteristics of neonatal platelet reactivity. Pharmacol. Res. 123, 1–9 (2017).

    Article  CAS  PubMed  Google Scholar 

  15. Davenport, P. & Sola‐Visner, M. Platelets in the neonate: not just a small adult. Res Pr. Thromb. Haemost. 6, e12719 (2022).

    Article  CAS  Google Scholar 

  16. Urban, D. et al. Decreased numbers of dense granules in fetal and neonatal platelets. Haematologica 102, e36–e38 (2017).

    Article  PubMed  PubMed Central  Google Scholar 

  17. Hvas, A.-M. & Favaloro, E. J. Platelet function testing in pediatric patients. Expert Rev. Hematol. 10, 281–288 (2017).

    Article  CAS  PubMed  Google Scholar 

  18. Yip, C. & García, Á. Exploring the potential of platelet proteomics in children. Prot. Clin. Appl. 8, 807–812 (2014).

    Article  CAS  Google Scholar 

  19. Stokhuijzen, E. et al. Differences between platelets derived from neonatal cord blood and adult peripheral blood assessed by mass spectrometry. J. Proteome Res. 16, 3567–3575 (2017).

    Article  CAS  PubMed  Google Scholar 

  20. Sveshnikova, A. N. et al. Systems biology insights into the meaning of the platelet’s dual-receptor thrombin signaling. J. Thromb. Haemost. 14, 2045–2057 (2016).

    Article  CAS  PubMed  Google Scholar 

  21. Best, M. G. et al. RNA-Seq of tumor-educated platelets enables blood-based pan-cancer, multiclass, and molecular pathway cancer diagnostics. Cancer Cell 28, 666–676 (2015).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Schwanhäusser, B. et al. Global quantification of mammalian gene expression control. Nature 473, 337–342 (2011).

    Article  ADS  PubMed  Google Scholar 

  23. Tyanova, S., Temu, T. & Cox, J. The MaxQuant computational platform for mass spectrometry-based shotgun proteomics. Nat. Protoc. 11, 2301–2319 (2016).

    Article  CAS  PubMed  Google Scholar 

  24. Farrah, T. et al. A high-confidence human plasma proteome reference set with estimated concentrations in PeptideAtlas. Mol. Cell Proteom. 10, M110.006353 (2011).

    Article  Google Scholar 

  25. Bryk, A. H. & Wiśniewski, J. R. Quantitative analysis of human red blood cell proteome. J. Proteome Res 16, 2752–2761 (2017).

    Article  CAS  PubMed  Google Scholar 

  26. Martyanov, A. A. et al. Heterogeneity of Integrin αIIbβ3 function in pediatric immune thrombocytopenia revealed by continuous flow cytometry analysis. Int J. Mol. Sci. 21, E3035 (2020).

    Article  MathSciNet  Google Scholar 

  27. Grynkiewicz, G., Poenie, M. & Tsien, R. Y. A new generation of Ca2+ indicators with greatly improved fluorescence properties. J. Biol. Chem. 260, 3440–3450 (1985).

    Article  CAS  PubMed  Google Scholar 

  28. Martyanov, A. A. et al. Longitudinal multiparametric characterization of platelet dysfunction in COVID-19: effects of disease severity, anticoagulation therapy and inflammatory status. Thromb. Res 211, 27–37 (2022).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Burkhart, J. M. et al. The first comprehensive and quantitative analysis of human platelet protein composition allows the comparative analysis of structural and functional pathways. Blood 120, e73–e82 (2012).

    Article  CAS  PubMed  Google Scholar 

  30. Stalker, T. J. Platelet activation gradients during thrombus formation. Blood 126, SCI-13 (2015).

    Article  Google Scholar 

  31. Colberg, L., Cammann, C., Greinacher, A. & Seifert, U. Structure and function of the ubiquitin‐proteasome system in platelets. J. Thromb. Haemost. 18, 771–780 (2020).

    Article  CAS  PubMed  Google Scholar 

  32. El-Kadiry, A. E.-H. & Merhi, Y. The role of the proteasome in platelet function. Int. J. Mol. Sci. 22, 3999 (2021).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Montague, S. J., Andrews, R. K. & Gardiner, E. E. Mechanisms of receptor shedding in platelets. Blood 132, 2535–2545 (2018).

    Article  CAS  PubMed  Google Scholar 

  34. Huang, J. et al. Molecular proteomics and signalling of human platelets in health and disease. IJMS 22, 9860 (2021).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Tassi Yunga, S. et al. Effects of ex vivo blood anticoagulation and preanalytical processing time on the proteome content of platelets. J. Thromb. Haemost. 20, 1437–1450 (2022).

    Article  CAS  PubMed  Google Scholar 

  36. Lehallier, B. et al. Undulating changes in human plasma proteome profiles across the lifespan. Nat. Med. 25, 1843–1850 (2019).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. Eshel-Green, T., Berny, M., Conley, R. & McCarty, O. Effect of sex difference on platelet adhesion, spreading and aggregate formation under flow. Thromb. Haemost. 102, 958–965 (2009).

    Article  CAS  PubMed  Google Scholar 

  38. Soo Kim, B., et al. Sex-specific platelet activation through protease-activated receptors reverses in myocardial infarction. ATVB https://doi.org/10.1161/ATVBAHA.120.315033 (2020).

  39. Jayachandran, M. et al. Sex-specific changes in platelet aggregation and secretion with sexual maturity in pigs. J. Appl Physiol. (1985) 97, 1445–1452 (2004).

    Article  CAS  PubMed  Google Scholar 

  40. Cini, C. et al. Differences in the resting platelet proteome and platelet releasate between healthy children and adults. J. Proteom. 123, 78–88 (2015).

    Article  CAS  Google Scholar 

  41. Luo, S.-Z., Mo, X., López, J. A. & Li, R. Role of the transmembrane domain of glycoprotein IX in assembly of the glycoprotein Ib-IX complex. J. Thromb. Haemost. 5, 2494–2502 (2007).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  42. Mo, X., Liu, L., López, J. A. & Li, R. Transmembrane domains are critical to the interaction between platelet glycoprotein V and glycoprotein Ib-IX complex: GPV and GPIb-IX interaction. J. Thromb. Haemost. 10, 1875–1886 (2012).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  43. Modderman, P. W., Admiraal, L. G., Sonnenberg, A., von dem Borne, A. E. Glycoproteins V and Ib-IX form a noncovalent complex in the platelet membrane. J. Biol. Chem. 267, 364–369 (1992).

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Funding

Platelet proteomics assays (Figs. 1, 2, Supplementary materials) were funded by Russian Science Foundation, grant number 21-74-20087; proteomics data analysis (Fig. 3) was funded by Russian Science Foundation, grant number 23-74-00057; platelet functional and signaling analysis (Figs. 4, 5) was funded by the endowment foundation “Science for Children”.

Author information

Authors and Affiliations

  1. Center for Theoretical Problems of Physico-Сhemical Pharmacology, Russian Academy of Sciences, 30 Srednyaya Kalitnikovskaya str., Moscow, 109029, Russia

    Andrei K. Garzon Dasgupta, Alexey A. Martyanov, Anastasia A. Ignatova, Mikhail A. Panteleev & Anastasia N. Sveshnikova

  2. National Medical Research Centеr of Pediatric Hematology, Oncology and Immunology named after Dmitry Rogachev, 1 Samory Mashela St, Moscow, 117198, Russia

    Alexey A. Martyanov, Anastasia A. Ignatova, Galina A. Novichkova, Mikhail A. Panteleev & Anastasia N. Sveshnikova

  3. Institute of Biomedical Chemistry, 10 bld. 8, Pogodinskaya str., 119121, Moscow, Russia

    Victor G. Zgoda

  4. Lomonosov Moscow State University, 1/2 Leninskie gory, Moscow, 119991, Russia

    Mikhail A. Panteleev & Anastasia N. Sveshnikova

Authors
  1. Andrei K. Garzon Dasgupta
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  2. Alexey A. Martyanov
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  3. Anastasia A. Ignatova
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  7. Anastasia N. Sveshnikova
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Contributions

A.K.G.D. analyzed data, prepared samples for proteomics analysis, and wrote text; A.A.M. performed intracellular signaling assays, prepared samples for proteomics, analyzed flow cytometry data, wrote the text and edited the paper; A.A.I. performed platelet functional testing; V.G.Z. performed proteomics assays and analyzed proteomics data; G.A.N. analysed hematological status of the donors and edited the paper; M.A.P. analyzed proteomics and platelet functional data; A.N.S. recruited the resources for the study, conceptualized the project, analyzed the data, edited the paper and supervised the study.

Corresponding author

Correspondence to Anastasia N. Sveshnikova.

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The authors declare no competing interests.

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The study was conducted in accordance with the Declaration of Helsinki, and approved by the Institutional Review Board (or Ethics Committee) of CTP PCP RAS (protocol code 2 06.04.2021)

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Informed consent was obtained from each of the included donors or their legal representatives.

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Garzon Dasgupta, A.K., Martyanov, A.A., Ignatova, A.A. et al. Comparison of platelet proteomic profiles between children and adults reveals origins of functional differences. Pediatr Res 95, 966–973 (2024). https://doi.org/10.1038/s41390-023-02865-y

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