Healthy pediatric platelets are moderately hyporeactive in comparison with adults’ platelets

Пономаренко, Е А; Ignatova, Anastasia A.; Polokhov, Dmitry M.; Khismatullina, Rimma; Курило, Д. С.; Shcherbina, Anna; Zharkov, P. A.; Maschan, Alexey; Novichkova, Galina; Panteleev, Mikhail A.

doi:10.1080/09537104.2021.1981848

Cited by 5 publications

(9 citation statements)

References 23 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…These differences were observed in response to collagen or Thr-receptor agonists like CRP, SFLLRN and AYPGKF, but not in response to ADP. 6 In line with this, we have previously described an impaired CLEC-2 and GPVI activation in neonatal platelets. 8 Interestingly, dense granule content was described to be reduced in neonatal platelets, when observed by electron microscopy, but this was not corroborated by other research group using different experimental techniques.…”

supporting

confidence: 62%

“…Additionally, incubation of platelets with the specific PAR-4 agonist, AYPGKF, failed to induce significant changes in cytosolic Ca 2+ concentration, which is in agreement with previous studies. 6,8,39 Discrepancies in the PAR expression and function may explain the observed differences in Ca 2+ mobilization in response to Thr in neonatal platelets, which has been previously reported. 26 Interestingly, reduced Thr-evoked Ca 2+ mobilization was found in maternal platelets compared with platelets from control women, which may be explained by alteration in PAR-1-agonist, as evidence using SFLLRN; nonetheless, PAR-1-mediated activation of Ca 2+ entry appears not to be altered in maternal platelets, in contrast to the alteration found in neonatal platelets.…”

Section: Discussionmentioning

confidence: 75%

“…Neonatal platelets show alterations downstream of several platelet receptors (CLEC-2, GPVI and PAR-4 6,8 ), leading to reduced platelet activation. In fact, aggregation in neonatal platelets is altered in response to collagen, ATP and Thr, as shown in this and previous studies performed by others.…”

Section: Discussionmentioning

confidence: 99%

“…Among others, neonatal platelets present a reduced response to ADP, epinephrine, collagen, thrombin (Thr) and thromboxane analogues (TXA 2 ). [5][6][7] A very recent study has reported differences between neonatal and maternal…”

mentioning

confidence: 99%

See 3 more Smart Citations

SARAF overexpression impairs thrombin‐induced Ca²⁺ homeostasis in neonatal platelets

Berna‐Erro,

Granados,

Teruel‐Montoya

et al. 2023

Br J Haematol

View full text Add to dashboard Cite

SummaryNeonatal platelets present a reduced response to the platelet agonist, thrombin (Thr), thus resulting in a deficient Thr‐induced aggregation. These alterations are more pronounced in premature newborns. Here, our aim was to uncover the causes underneath the impaired Ca2+ homeostasis described in neonatal platelets. Both Ca2+ mobilization and Ca2+ influx in response to Thr are decreased in neonatal platelets compared to maternal and control woman platelets. In neonatal platelets, we observed impaired Ca2+ mobilization in response to the PAR‐1 agonist (SFLLRN) or by blocking SERCA3 function with tert‐butylhydroquinone. Regarding SOCE, the STIM1 regulatory protein, SARAF, was found overexpressed in neonatal platelets, promoting an increase in STIM1/SARAF interaction even under resting conditions. Additionally, higher interaction between SARAF and PDCD61/ALG2 was also observed, reducing SARAF ubiquitination and prolonging its half‐life. These results were reproduced by overexpressing SARAF in MEG01 and DAMI cells. Finally, we also observed that pannexin 1 permeability is enhanced in response to Thr in control woman and maternal platelets, but not in neonatal platelets, hence, leading to the deregulation of the Ca2+ entry found in neonatal platelets. Summarizing, we show that in neonatal platelets both Ca2+ accumulation in the intracellular stores and Thr‐evoked Ca2+ entry through either capacitative channels or non‐selective channels are altered in neonatal platelets, contributing to deregulated Ca2+ homeostasis in neonatal platelets and leading to the altered aggregation observed in these subjects.

show abstract

supporting

confidence: 62%

Section: Discussionmentioning

confidence: 75%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

See 2 more Smart Citations

SARAF overexpression impairs thrombin‐induced Ca²⁺ homeostasis in neonatal platelets

Berna‐Erro,

Granados,

Teruel‐Montoya

et al. 2023

Br J Haematol

View full text Add to dashboard Cite

show abstract

“…[1][2][3][4] Specific agonists of protease activated receptors 1 (PAR1) and 4 (PAR4) are often added to mimic the physiological activation by thrombin. [5][6][7][8][9][10] These compounds are utilized most often to assess platelet activity in whole blood, platelet-rich plasma or isolated platelet suspensions. 1 The results obtained under different experimental conditions are often extrapolated across models.…”

Section: Introductionmentioning

confidence: 99%

C1-inhibitor influence on platelet activation by thrombin receptors agonists

Tarandovskiy

Buehler

Karnaukhova

2022

Clin Appl Thromb Hemost

View full text Add to dashboard Cite

Introduction Protease activated receptors 1 (PAR1) and 4 (PAR4) agonists are used to study platelet activation. Data on platelet activation are extrapolated across experimental settings. C1-inhibitor (C1INH) is a protease inhibitor present in plasma but not in isolated platelet suspensions. Here we show that C1INH affects platelet activation through PAR1 and PAR4 agonists. Methods Platelets were isolated from healthy donor whole blood and then labeled with anti-CD62P and PAC1 antibodies. The platelet suspensions were exposed to PAR1 agonists SFLLRN, TFLLR and TFLLRN; PAR4 agonists AYPGKF and GYPGQV; ADP and thrombin. Flow-cytometric measurements were performed in 5, 10 and 15 min after activation. Results 0.25 mg/ml C1INH addition made platelets to faster expose CD62P and glycoprotein IIb/IIIa complex after activation with PAR1 agonists. Conversely, C1INH addition led to inhibition of platelet activation with PAR4 agonists and thrombin. Activation with ADP was not affected by C1INH. Conclusions Our results suggest that C1INH can modify platelet activation in the presence of synthetic PAR agonists used in platelet research. These observations may be relevant to the development of new methods to assess platelet function.

show abstract

Flow cytometry for comprehensive assessment of platelet functional activity in response to ADP stimulation

Ponomarenko,

Ignatova,

Polokhov

et al. 2023

European J of Haematology

Self Cite

View full text Add to dashboard Cite

ObjectivesFlow cytometry with adenosine diphosphate (ADP) allows to characterize molecular changes of platelet function caused by this physiologically important activation, but the methodology has not been thoroughly investigated, standardized and characterized yet. We analyzed the influence of several major variables and chose optimal conditions for platelet function assessment.MethodsFor activation, 2.5 μM CaCl2, 5 μM ADP and antibodies were added to diluted blood and incubated for 15 min. We analyzed kinetics of antibody binding and effects of their addition sequence, agonist concentration, blood dilution, exogenous calcium addition and platelet fixation.ResultsWe tested our protocol on 11 healthy children, 22 healthy adult volunteers, 9 patients after a month on dual antiplatelet therapy after percutaneous coronary intervention (PCI), 7 adult patients and 14 children with immune thrombocytopenia (ITP). We found that our protocol is highly sensitive to ADP stimulation with low percentage of aggregates formation. The assay is also sensitive to platelet function inhibition in post‐PCI patients. Finally, platelet preactivation with ITP plasma was stronger and caused increase in activation response to ADP stimulation compared to preactivation with low dose of ADP.ConclusionsOur assay is sensitive to antiplatelet therapy and platelet preactivation in ITP patients under physiological conditions with minimal percentage of aggregates formation.

show abstract

Healthy pediatric platelets are moderately hyporeactive in comparison with adults’ platelets

Cited by 5 publications

References 23 publications

SARAF overexpression impairs thrombin‐induced Ca²⁺ homeostasis in neonatal platelets

SARAF overexpression impairs thrombin‐induced Ca²⁺ homeostasis in neonatal platelets

C1-inhibitor influence on platelet activation by thrombin receptors agonists

Flow cytometry for comprehensive assessment of platelet functional activity in response to ADP stimulation

Contact Info

Product

Resources

About

Healthy pediatric platelets are moderately hyporeactive in comparison with adults’ platelets

Cited by 5 publications

References 23 publications

SARAF overexpression impairs thrombin‐induced Ca2+ homeostasis in neonatal platelets

SARAF overexpression impairs thrombin‐induced Ca2+ homeostasis in neonatal platelets

C1-inhibitor influence on platelet activation by thrombin receptors agonists

Flow cytometry for comprehensive assessment of platelet functional activity in response to ADP stimulation

Contact Info

Product

Resources

About

SARAF overexpression impairs thrombin‐induced Ca²⁺ homeostasis in neonatal platelets

SARAF overexpression impairs thrombin‐induced Ca²⁺ homeostasis in neonatal platelets