Protective role of Cav-1 in pneumolysin-induced endothelial barrier dysfunction.
Protective role of Cav-1 in pneumolysin-induced endothelial barrier dysfunction.
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DOI：
10.3389/fimmu.2022.945656
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发表时间：
2022
影响因子：
7.3
通讯作者：
中科院分区：
文献类型：
--
作者：
肺炎链球菌溶血素（Streptococcus pneumonia，EAE）是肺炎链球菌的主要致病因子。胆固醇结合内皮细胞（EC）质膜的富含胆固醇的结构域，导致孔组装和细胞内（IC）Ca 2+水平升高，损害内皮屏障完整性。小窝是富含胆固醇的EC的专门质膜微区。我们假设EC质膜中富含胆固醇的结构域的丰度赋予细胞对胆固醇的易感性。与这一假设相反，我们发现增加PLY诱导的IC Ca 2+膜胆固醇耗竭后。Caveolin-1（Cav-1）是Caveolae的重要结构蛋白，其表达受胆固醇水平的调节，提示其可能在EC屏障功能中发挥作用。事实上，Cav-1及其支架结构域肽保护内皮屏障免受PLY诱导的破坏。在功能丧失实验中，使用CRISPR-Cas9敲除Cav-1或在人肺微血管EC中沉默Cav-1。Cav-1的缺失显著增强了CaV破坏内皮屏障完整性的能力。再表达Cav-1或其支架结构域肽的救援实验保护EC屏障免受PLY诱导的屏障破坏。动力蛋白-2（DNM 2）已知调节小窝膜内吞作用。用发动蛋白抑制剂或siDNM 2扩增的抑制剂抑制内吞作用诱导EC屏障功能障碍。这些结果表明Cav-1通过促进受损膜的内吞作用，从而减少钙进入和PLY依赖性信号传导，保护内皮屏障免受损伤。
Pneumolysin (PLY) is a bacterial pore forming toxin and primary virulence factor of Streptococcus pneumonia, a major cause of pneumonia. PLY binds cholesterol-rich domains of the endothelial cell (EC) plasma membrane resulting in pore assembly and increased intracellular (IC) Ca2+ levels that compromise endothelial barrier integrity. Caveolae are specialized plasmalemma microdomains of ECs enriched in cholesterol. We hypothesized that the abundance of cholesterol-rich domains in EC plasma membranes confers cellular susceptibility to PLY. Contrary to this hypothesis, we found increased PLY-induced IC Ca2+ following membrane cholesterol depletion. Caveolin-1 (Cav-1) is an essential structural protein of caveolae and its regulation by cholesterol levels suggested a possible role in EC barrier function. Indeed, Cav-1 and its scaffolding domain peptide protected the endothelial barrier from PLY-induced disruption. In loss of function experiments, Cav-1 was knocked-out using CRISPR-Cas9 or silenced in human lung microvascular ECs. Loss of Cav-1 significantly enhanced the ability of PLY to disrupt endothelial barrier integrity. Rescue experiments with re-expression of Cav-1 or its scaffolding domain peptide protected the EC barrier against PLY-induced barrier disruption. Dynamin-2 (DNM2) is known to regulate caveolar membrane endocytosis. Inhibition of endocytosis, with dynamin inhibitors or siDNM2 amplified PLY induced EC barrier dysfunction. These results suggest that Cav-1 protects the endothelial barrier against PLY by promoting endocytosis of damaged membrane, thus reducing calcium entry and PLY-dependent signaling.
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影响因子：
29
作者：
Fernández-Hernando C;Yu J;Suárez Y;Rahner C;Dávalos A;Lasunción MA;Sessa WC
通讯作者：
Sessa WC
影响因子：
3.7
作者：
Byrne DP;Dart C;Rigden DJ
通讯作者：
Rigden DJ
影响因子：
3.4
作者：
Gilbert, Robert J. C.
通讯作者：
Gilbert, Robert J. C.
影响因子：
6.7
作者：
Greene NG;Narciso AR;Filipe SR;Camilli A
通讯作者：
Camilli A
RhoA S-亚硝基化作为调节机制影响内皮屏障功能以响应 G-细菌毒素。
影响因子：
5.8
作者：
Chen F;Wang Y;Rafikov R;Haigh S;Zhi WB;Kumar S;Doulias PT;Rafikova O;Pillich H;Chakraborty T;Lucas R;Verin AD;Catravas JD;She JX;Black SM;Fulton DJR
通讯作者：
Fulton DJR