Platelet WDR1 suppresses platelet activity and is associated with cardiovascular disease
Platelet WDR1 suppresses platelet activity and is associated with cardiovascular disease
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DOI：
10.1182/blood-2016-03-703157
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发表时间：
2016-10-20
期刊：
影响因子：
20.3
通讯作者：
Berger, Jeffrey S.
中科院分区：
文献类型：
--
作者：
Montenont, Emilie;Echagarruga, Christina;Berger, Jeffrey S.
血小板活性在止血中起主要作用，血小板活性升高可能导致动脉粥样硬化血栓形成。我们试图在健康对照中确定血小板活性变异性和血小板相关基因之间的关联。血小板转录谱分析显示，高反应性血小板表型受试者血小板信使RNA （mRNA）中的WD-40重复结构域1 （WDR1）下调，WDR1是肌动蛋白解聚合因子活性的增强子。我们使用人巨核母细胞系MEG-01作为人巨核细胞和血小板的体外模型。凝血酶刺激MEG-01可降低WDR1转录本和蛋白水平。在MEG-01细胞中，WDR1敲低（KD）增加了基础状态和激活状态下的粘附和扩散，增加了F-actin含量，增加了基础细胞内钙浓度。WDR1 KD细胞产生的血小板样颗粒（PLPs）比未修饰的mg -01细胞产生的PLPs数量少，但体积大，并且在基础状态和凝血酶激活时具有显著增加的粘附性。相反，WDR1过表达逆转了巨核细胞和PLPs的WDR1 KD表型。为了解释这些发现的临床意义，我们测量了患有心血管疾病的受试者（n = 27）和年龄和性别匹配的对照组（n = 10）的血小板RNA中WDR1的表达。心血管疾病患者的WDR1 mRNA和蛋白水平明显降低。这些数据表明，WDR1在抑制血小板活性中起重要作用，它改变肌动蛋白细胞骨架动力学，WDR1的下调可能有助于血小板介导的心血管疾病发病机制。
Platelet activity plays a major role in hemostasis with increased platelet activity likely contributing to the pathogenesis of atherothrombosis. We sought to identify associations between platelet activity variability and platelet-related genes in healthy controls. Transcriptional profiling of platelets revealed that WD-40 repeat domain 1 (WDR1), an enhancer of actin-depolymerizing factor activity, is downregulated in platelet messenger RNA (mRNA) from subjects with a hyperreactive platelet phenotype. We used the human megakaryoblastic cell line MEG-01 as an in vitro model for human megakaryocytes and platelets. Stimulation of MEG-01 with thrombin reduced levels of WDR1 transcripts and protein. WDR1 knockdown (KD) in MEG-01 cells increased adhesion and spreading in both the basal and activated states, increased F-actin content, and increased the basal intracellular calcium concentration. Platelet-like particles (PLPs) produced by WDR1 KD cells were fewer in number but larger than PLPs produced from unmodified MEG-01 cells, and had significantly increased adhesion in the basal state and upon thrombin activation. In contrast, WDR1 overexpression reversed the WDR1 KD phenotype of megakaryocytes and PLPs. To translate the clinical significance of these findings, WDR1 expression was measured in platelet RNA from subjects with established cardiovascular disease (n = 27) and age-and sex-matched controls (n = 10). The WDR1 mRNA and protein level was significantly lower in subjects with cardiovascular disease. These data suggest that WDR1 plays an important role in suppressing platelet activity, where it alters the actin cytoskeleton dynamics, and downregulation of WDR1 may contribute to the platelet-mediated pathogenesis of cardiovascular disease.