Department of Laboratory Medicine (S.X.G.), Yale University School of Medicine, New Haven, CT.
Department of Pediatrics (B.S.M., E.V.S.F.), Yale University School of Medicine, New Haven, CT.
Arterioscler Thromb Vasc Biol. 2024 Dec;44(12):2530-2539. doi: 10.1161/ATVBAHA.124.321131. Epub 2024 Aug 22.
Congenital heart disease (CHD) is a group of complex heart defects associated with hematologic abnormalities, including increased risk of thrombotic and bleeding events. Past studies have observed evidence of platelet hyperreactivity, while other studies showed decreased platelet activation in patients with CHD. The goal of this study was to develop a mass spectrometry approach to characterize single platelets in infants with CHD and identify potential etiology for such discrepant results.
We enrolled 19 infants with CHD along with 21 non-CHD controls at Yale New Haven Children's Heart Center. A single-cell high-dimensional mass cytometry method was developed to quantitatively interrogate platelet surface markers in whole blood. Additionally, plasma cytokine analysis was performed through a multiplexed panel of 52 vascular and inflammatory markers to assess for platelet releasates.
We found that infants with CHD had significant differences in platelet activation and functional markers by mass cytometry compared with non-CHD controls. Based on cell surface markers, we classified the platelets into 8 subpopulations (P0 to P7). Distinct subpopulations of platelets (P1, P4, and P5) exhibiting decreased aggregatory phenotype but altered secretory phenotypes were also identified and found to be more abundant in the blood of infants with CHD. Electron microscopy identified increased proportion of hypogranular platelets in CHD. Moreover, cytokine analysis demonstrated an overall increase in plasma cytokines and biomarkers in CHD, including IL (interleukin)-6, IL-8, IL-27, RANTES (regulated upon activation, normal T cell expressed and secreted), and VWF (von Willebrand factor), which are expressed in platelet granules and can be released upon activation.
We developed a robust mass cytometry approach to identify platelet phenotypic heterogeneity. Infants with CHD had alterations in distinct subpopulations of platelets with overall reduced aggregatory phenotype and secretory dysfunction. These findings suggest that platelets in infants with CHD may be exhausted due to persistent stimulation and may explain both bleeding and thrombotic vascular complications associated with CHD.
先天性心脏病（CHD）是一组与血液异常相关的复杂心脏缺陷，包括血栓形成和出血事件的风险增加。过去的研究已经观察到血小板高反应性的证据，而其他研究则显示 CHD 患者的血小板活化减少。本研究的目的是开发一种质谱分析方法来描述 CHD 婴儿的单个血小板，并确定导致这种不一致结果的潜在病因。
我们在耶鲁纽黑文儿童心脏中心招募了 19 名患有 CHD 的婴儿和 21 名非 CHD 对照。开发了一种单细胞高维质谱细胞术方法来定量检测全血中的血小板表面标志物。此外，通过 52 种血管和炎症标志物的多重分析面板进行血浆细胞因子分析，以评估血小板释放物。
与非 CHD 对照组相比，我们发现 CHD 婴儿的血小板活化和功能标志物通过质谱细胞术有显著差异。基于细胞表面标志物，我们将血小板分为 8 个亚群（P0 至 P7）。还鉴定了具有降低聚集表型但改变分泌表型的血小板独特亚群（P1、P4 和 P5），并且在 CHD 婴儿的血液中发现它们更为丰富。电子显微镜鉴定出 CHD 中颗粒较少的血小板比例增加。此外，细胞因子分析表明 CHD 中血浆细胞因子和生物标志物总体增加，包括白细胞介素（IL）-6、IL-8、IL-27、RANTES（激活正常 T 细胞表达和分泌）和 VWF（血管性血友病因子），它们在血小板颗粒中表达并可在活化时释放。
我们开发了一种强大的质谱细胞术方法来鉴定血小板表型异质性。CHD 婴儿的血小板存在不同亚群的改变，总体聚集表型降低和分泌功能障碍。这些发现表明，CHD 婴儿的血小板可能由于持续刺激而耗尽，并可能解释与 CHD 相关的出血和血栓性血管并发症。
