Evaluation of targeting c-Src by the RGT-containing peptide as a novel antithrombotic strategy.
Evaluation of targeting c-Src by the RGT-containing peptide as a novel antithrombotic strategy.
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含 RGT 的肽靶向 c-Src 作为新型抗血栓策略的评估。
DOI：
10.1186/s13045-015-0159-8
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发表时间：
2015-05-30
影响因子：
28.5
通讯作者：
Xi X
中科院分区：
文献类型：
--
作者：
Huang J;Shi X;Xi W;Liu P;Long Z;Xi X
整合素β3与c-Src的相互作用在细胞信号传导中起关键作用，其在血小板粘附和聚集以及肿瘤细胞增殖和转移或骨细胞骨吸收中有重要意义。选择性阻断血小板中整合素αIIbβ3由外向内的信号传导已成为关注的焦点，因为其有效的抗血栓形成潜力以及足够的止血能力。豆蔻酰化RGT肽已被证明通过靶向c-Src与整联蛋白β3尾的结合来实现这种阻断，但缺乏关于作用机制的关键信息阻止了该策略进一步发展为实用的抗血栓形成剂。因此，需要深入了解RGT肽在调节血小板功能中的确切机制，以建立通过靶向c-Src的潜在抗血栓治疗的基础。应用还原敏感肽以排除胞质递送后的膜锚定。在活细胞或蛋白质水平测定c-Src活性，以评估RGT靶向对c-Src的直接影响。通过将全血灌注通过胶原包被的微室，观察在细胞质RGT肽存在下的流动下血栓形成。RGT肽不依赖于膜锚定来抑制血小板中的由外向内信号传导。myr-AC ~ CRGT肽通过破坏Src/β3结合而容易阻断激动剂诱导的c-Src活化，并且除了典型的由外向内信号传导事件之外，还抑制RhoA活化和胶原诱导的血小板聚集。myr-AC ~ CRGT对活细胞中c-Src的激酶活性没有直接影响，这可以通过其不能使Csk与c-Src解离或不能改变c-Src Y 416和Y 527的磷酸化水平来证明，体外激酶测定的结果也是一致的。在流动条件下，myr-AC ~ CRGT肽主要在高剪切速率下引起血小板血栓形成的抑制。这些发现为RGT肽调节整联蛋白信号传导和血小板功能的分子机制提供了新的见解，并增强了RGT肽诱导的Src/β3结合破坏作为可药用靶点的潜力，最终可以使用基于结构的小分子模拟物进行体内和临床研究。本文的在线版本（doi：10.1186/s13045-015-0159-8）包含补充材料，可供授权用户使用。
Interaction of integrin β3 with c-Src plays critical roles in cellular signaling which is heavily implicated in platelet adhesion and aggregation, as well as in tumor cell proliferation and metastasis or in osteoclastic bone resorption. Selectively blocking integrin αIIbβ3 outside-in signaling in platelets has been a focus of attention because of its effective antithrombotic potential together with a sufficient hemostatic capacity. The myristoylated RGT peptide has been shown to achieve this blockade by targeting the association of c-Src with the integrin β3 tail, but the lack of key information regarding the mechanisms of action prevents this strategy from being further developed into practical antithrombotics. Therefore, in-depth knowledge of the precise mechanisms for RGT peptide in regulating platelet function is needed to establish the basis for a potential antithrombotic therapy by targeting c-Src. The reduction-sensitive peptides were applied to rule out the membrane anchorage after cytoplasmic delivery. The c-Src activity was assayed at living cell or at protein levels to assess the direct effect of RGT targeting on c-Src. Thrombus formation under flow in the presence of cytoplasmic RGT peptide was observed by perfusing whole blood through the collagen-coated micro-chamber. The RGT peptide did not depend on the membrane anchorage to inhibit outside-in signaling in platelets. The myr-AC ~ CRGT peptide readily blocked agonist-induced c-Src activation by disrupting the Src/β3 association and inhibited the RhoA activation and collagen-induced platelet aggregation in addition to the typical outside-in signaling events. The myr-AC ~ CRGT had no direct effect on the kinase activity of c-Src in living cells as evidenced by its inability to dissociate Csk from c-Src or to alter the phosphorylation level of c-Src Y416 and Y527, consistent results were also from in vitro kinase assays. Under flow conditions, the myr-AC ~ CRGT peptide caused an inhibition of platelet thrombus formation predominantly at high shear rates. These findings provide novel insights into the molecular mechanisms by which the RGT peptide regulates integrin signaling and platelet function and reinforce the potential of the RGT peptide-induced disruption of Src/β3 association as a druggable target that would finally enable in vivo and clinical studies using the structure-based small molecular mimetics. The online version of this article (doi:10.1186/s13045-015-0159-8) contains supplementary material, which is available to authorized users.
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影响因子：
--
作者：
Huveneers S;Danen EH
通讯作者：
Danen EH
影响因子：
10.4
作者：
Rossaint, J.;Vestweber, D.;Zarbock, A.
通讯作者：
Zarbock, A.
影响因子：
82.9
作者：
Nesbitt, Warwick S.;Westein, Erik;Jackson, Shaun P.
通讯作者：
Jackson, Shaun P.
DOI：
10.1111/j.1538-7836.2009.03581.x
发表时间：
2009-11
期刊：
Journal of thrombosis and haemostasis : JTH
影响因子：
--
作者：
Naik MU;Nigam A;Manrai P;Millili P;Czymmek K;Sullivan M;Naik UP
通讯作者：
Naik UP
影响因子：
20.3
作者：
Goschnick, Matt W.;Lau, Lai-Man;Jackson, Denise E.
通讯作者：
Jackson, Denise E.