Trpc6 gain-of-function disease mutation enhances phosphatidylserine exposure in murine platelets.
Trpc6 gain-of-function disease mutation enhances phosphatidylserine exposure in murine platelets.
复制标题
DOI：
10.1371/journal.pone.0270431
复制
发表时间：
2022
期刊：
影响因子：
3.7
通讯作者：
Schlondorff, Johannes S.
中科院分区：
文献类型：
--
作者：
Boekell, Kimber L.;Brown, Brittney J.;Talbot, Brianna E.;Schlondorff, Johannes S.
血小板通过暴露其细胞表面上的磷脂酰丝氨酸（PS）以响应强激动剂活化来增强凝血。瞬时受体电位通道，包括TRPC 6，已被牵连的钙内流中央到这个过程。在这里，我们描述了Trpc 6功能获得性（GOF）疾病相关和显性阴性（DN）突变对小鼠血小板活化的影响。对来自携带Trpc 6 E896 K/E896 K（GOF）和Trpc 6DN/DN突变的小鼠的血小板进行体外分析。在加入TRPC 3/6激动剂GSK 1702934 A（GSK）后，Trpc 6 E896 K/E896 K和Trpc 6DN/DN突变体血小板分别显示增强的和缺乏的钙内流。GSK足以诱导Trpc 6 E896 K/E896 K中整合素αIIbβ3活化、P-选择和PS暴露、talin裂解和MLC 2磷酸化，但在野生型血小板中不诱导。凝血酶诱导的钙内流和PS暴露增强，凝块收缩延迟，由GOF TRPC 6，而野生型和Trpc 6DN/DN血小板之间没有差异。相反，与野生型相比，GSK处理后的Erk活化在Trpc 6DN/DN中不存在，并且在Trpc 6 E896 K/E896 K血小板中增强。正变构调节剂TRPC 6-PAM-C20和氟西汀分别维持其增强和抑制Trpc 6 E896 K/E896 K血小板中GSK介导的钙内流的能力。数据表明，功能获得性突变体TRPC 6通道可以增强血小板活化，包括PS暴露，同时证实TRPC 6对于该过程不是必需的。此外，结果表明，Trpc 6 GOF疾病突变体并不简单地增加野生型TRPC 6反应，但可以影响通常不受TRPC 6通道活性调节的途径，显示出真正的功能获得性表型。
Platelets enhance coagulation by exposing phosphatidylserine (PS) on their cell surface in response to strong agonist activation. Transient receptor potential channels, including TRPC6, have been implicated in the calcium influx central to this process. Here, we characterize the effect of a Trpc6 gain-of-function (GOF) disease-associated, and a dominant negative (DN), mutation on murine platelet activation. Platelets from mice harboring Trpc6E896K/E896K (GOF) and Trpc6DN/DN mutations were subject to in vitro analysis. Trpc6E896K/E896K and Trpc6DN/DN mutant platelets show enhanced and absent calcium influx, respectively, upon addition of the TRPC3/6 agonist GSK1702934A (GSK). GSK was sufficient to induce integrin αIIbβ3 activation, P-selection and PS exposure, talin cleavage, and MLC2 phosphorylation in Trpc6E896K/E896K, but not in wild-type, platelets. Thrombin-induced calcium influx and PS exposure were enhanced, and clot retraction delayed, by GOF TRPC6, while no differences were noted between wild-type and Trpc6DN/DN platelets. In contrast, Erk activation upon GSK treatment was absent in Trpc6DN/DN, and enhanced in Trpc6E896K/E896K, platelets, compared to wild-type. The positive allosteric modulator, TRPC6-PAM-C20, and fluoxetine maintained their ability to enhance and inhibit, respectively, GSK-mediated calcium influx in Trpc6E896K/E896K platelets. The data demonstrate that gain-of-function mutant TRPC6 channel can enhance platelet activation, including PS exposure, while confirming that TRPC6 is not necessary for this process. Furthermore, the results suggest that Trpc6 GOF disease mutants do not simply increase wild-type TRPC6 responses, but can affect pathways not usually modulated by TRPC6 channel activity, displaying a true gain-of-function phenotype.
登录
查看更多内容
影响因子：
5.3
作者：
Dietrich, A;Schnitzler, MMY;Birnbaumer, L
通讯作者：
Birnbaumer, L
影响因子：
7.3
作者：
Harper, Matthew T.;Londono, Juan E. Camacho;Poole, Alastair W.
通讯作者：
Poole, Alastair W.
影响因子：
3.7
作者：
Dasgupta SK;Le A;Haudek SB;Entman ML;Rumbaut RE;Thiagarajan P
通讯作者：
Thiagarajan P
影响因子：
16.2
作者：
Guo, Wenjun;Tang, Qinglin;Chen, Lei
通讯作者：
Chen, Lei
影响因子：
9
作者：
Harper, M. T.;Poole, A. W.
通讯作者：
Poole, A. W.