A signaling pathway contributing to platelet storage lesion development: targeting PI3-kinase-dependent Rap1 activation slows storage-induced platelet deterioration.
A signaling pathway contributing to platelet storage lesion development: targeting PI3-kinase-dependent Rap1 activation slows storage-induced platelet deterioration.
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DOI：
10.1111/j.1537-2995.2009.02224.x
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发表时间：
2009-09
期刊：
影响因子：
2.9
通讯作者：
Kast J
中科院分区：
文献类型：
--
作者：
Schubert P;Thon JN;Walsh GM;Chen CH;Moore ED;Devine DV;Kast J
术语血小板贮存损伤（PSL）描述了血小板（PLT）在贮存期间的结构和生化变化。这些以形态特征和PLT代谢的改变为代表，导致功能降低，从而降低输血的活力。虽然表现的存储病变的特点，生化途径参与启动这一进程是未知的。最近，一种互补的蛋白质组学方法已被应用于分析储存过程中PLT蛋白质组的变化。通过采用严格的蛋白质组学标准，12种蛋白质被鉴定为在7天的储存期内相对浓度显著且一致地变化。使用显微镜、Western印迹分析、流式细胞术和PLT功能分析来阐明这12种蛋白质的亚组的参与，这些蛋白质通过整合素信号传导在一种潜在的储存损伤发展的信号传导途径中连接。显微镜分析显示，糖蛋白IIIa，Rap1，和talin在存储过程中的本地化的变化。观察到Rap1活化与PLT活化标志物CD62 P的表达相关。与PI 3激酶抑制剂LY294002孵育7天的PLT显示Rap1活化减少，整合素αIIbβ3活化和α颗粒释放中度减少。此外，这种抑制剂似乎在储存期间改善了PLT的完整性和质量，因为几种体外探针显示出PLT活化的减速。这些结果首次为PSL的信号通路介导提供了证据，其中PI3激酶依赖性Rap1激活导致整合素αIIbβ3激活和PLT脱粒。
The term platelet storage lesion (PSL) describes the structural and biochemical changes in platelets (PLTs) during storage. These are typified by alterations of morphologic features and PLT metabolism leading to reduced functionality and hence reduced viability for transfusion. While the manifestations of the storage lesion are well characterized, the biochemical pathways involved in the initiation of this process are unknown. A complementary proteomic approach has recently been applied to analyze changes in the PLT proteome during storage. By employing stringent proteomic criteria, 12 proteins were identified as significantly and consistently changing in relative concentration over a 7-day storage period. Microscopy, Western blot analysis, flow cytometry, and PLT functionality analyses were used to unravel the involvement of a subset of these 12 proteins, which are connected through integrin signaling in one potential signaling pathway underlying storage lesion development. Microscopic analysis revealed changes in localization of glycoprotein IIIa, Rap1, and talin during storage. Rap1 activation was observed to correlate with expression of the PLT activation marker CD62P. PLTs incubated for 7 days with the PI3-kinase inhibitor LY294002 showed diminished Rap1 activation as well as a moderate reduction in integrin αIIbβ3 activation and release of α-granules. Furthermore, this inhibitor seemed to improve PLT integrity and quality during storage as several in vitro probes showed a deceleration of PLT activation. These results provide the first evidence for a signaling pathway mediating PSL in which PI3-kinase–dependent Rap1 activation leads to integrin αIIbβ3 activation and PLT degranulation.