Tantiwong Chukiat, Cheung Hilaire Yam Fung, Dunster Joanne L, Gibbins Jonathan M, Heemskerk Johan W M, Cavill Rachel
Institute for Cardiovascular and Metabolic Research (ICMR), School of Biological Sciences, University of Reading, Whiteknights, Reading RG6 6AS, UK.
Department of Biochemistry, Maastricht University, P.O. Box 616, 6200 MD Maastricht, The Netherlands.
Int J Mol Sci. 2025 Jul 16;26(14):6820. doi: 10.3390/ijms26146820.
Platelets play critical roles in haemostasis and thrombosis. The platelet activation process is driven by agonist-induced rises in cytosolic [Ca], where the patterns of Ca responses are still incompletely understood. In this study, we developed a number of techniques to model the [Ca] curves of platelets from a single blood donor. Fura-2-loaded platelets were quasi-simultaneously stimulated with various agonists, i.e., thrombin, collagen, or CRP, in the presence or absence of extracellular Ca entry, secondary mediator effects, or Ca reuptake into intracellular stores. To understand the calibrated time curves of [Ca] rises, we developed two non-linear models, a multilayer perceptron (MLP) network and an autoregressive network with exogenous inputs (NARX). The trained networks accurately predicted the [Ca] curves for combinations of agonists and inhibitors, with the NARX model achieving an R of 0.64 for the trend prediction of unforeseen data. In addition, we used the same dataset for the construction of a partial least square (PLS) linear regression model, which estimated the explained variance of each input. The NARX model demonstrated that good fits could be obtained for the nanomolar [Ca] curves modelled, whereas the PLS model gave useful interpretable information on the importance of each variable. These modelling results can be used for the development of novel platelet [Ca]-inhibiting drugs, such as the drug 2-aminomethyl diphenylborinate, blocking Ca entry in platelets, or for the evaluation of general platelet signalling defects in patients with a bleeding disorder.
血小板在止血和血栓形成过程中发挥着关键作用。血小板激活过程由激动剂诱导的胞质[Ca]升高驱动，而Ca反应的模式仍未完全了解。在本研究中，我们开发了多种技术来模拟来自单一献血者的血小板的[Ca]曲线。用Fura-2负载的血小板在存在或不存在细胞外Ca内流、二级介质效应或Ca再摄取到细胞内储存的情况下，用各种激动剂（即凝血酶、胶原蛋白或CRP）进行准同时刺激。为了理解[Ca]升高的校准时间曲线，我们开发了两个非线性模型，一个多层感知器（MLP）网络和一个带有外部输入的自回归网络（NARX）。经过训练的网络准确地预测了激动剂和抑制剂组合的[Ca]曲线，NARX模型对意外数据的趋势预测的R值达到0.64。此外，我们使用相同的数据集构建了一个偏最小二乘（PLS）线性回归模型，该模型估计了每个输入的解释方差。NARX模型表明，对于所模拟的纳摩尔[Ca]曲线可以获得良好的拟合，而PLS模型给出了关于每个变量重要性的有用的可解释信息。这些建模结果可用于开发新型血小板[Ca]抑制药物，如药物2-氨基甲基二苯基硼酸酯，其可阻断血小板中的Ca内流，或用于评估出血性疾病患者的一般血小板信号缺陷。
