Effects of cognate, non-cognate and synthetic CXCR4 and ACKR3 ligands on human lung endothelial cell barrier function.
Effects of cognate, non-cognate and synthetic CXCR4 and ACKR3 ligands on human lung endothelial cell barrier function.
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DOI：
10.1371/journal.pone.0187949
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发表时间：
2017
期刊：
影响因子：
3.7
通讯作者：
Majetschak M
中科院分区：
文献类型：
--
作者：
Cheng YH;Eby JM;LaPorte HM;Volkman BF;Majetschak M
最近的证据表明，趋化因子 CXCL12（趋化因子受体 CXCR4 和 ACKR3 的同源激动剂）可减少凝血酶介导的内皮屏障功能损伤。目前尚缺乏 CXCL12 对凝血酶介导的人肺内皮通透性过高影响的详细表征，也无法获得结构-功能相关性。此外，其他 CXCR4/ACKR3 配体对肺内皮屏障功能的影响尚不清楚。因此，我们测试了一组 CXCR4/ACKR3 配体（CXCL12、CXCL11、泛素、AMD3100、TC14012）的作用，并比较了 CXCL12 变体（CXCL12α/β、CXCL12(3–68)、CXCL121、CXCL122、CXCL12-S-S4V、 CXCL12-R47E、CXCL12-K27A/R41A/R47A）及其在通透性测定中对人肺内皮屏障功能的影响。 CXCL12α增强人原代肺动脉内皮细胞（hPPAEC）屏障功能，而CXCL11、泛素、AMD3100和TC14012则无效。用 CXCL12α 和泛素预处理 hPPAEC 可降低凝血酶介导的通透性过高。凝血酶暴露后，CXCL12α-处理 hPPAEC 可将屏障功能损伤减少 70%（EC50 0.05-0.5nM），这可以被 AMD3100 拮抗；泛素（0.03-3μM）无效。在人肺微血管内皮细胞系 (HULEC5a) 中，CXCL12α 和泛素治疗后以类似程度减弱凝血酶诱导的通透性过高。在 Presto-Tango β-arrestin2 招募试验中，CXCL12(3–68) 无法有效激活 CXCR4； CXCL12-S-S4V、CXCL12-R47E 和 CXCL12-K27A/R41A/R47A 显示激活 CXCR4 的效力显着降低。虽然 ACKR3 Presto-Tango 测定中所有蛋白质的效力相当，但 CXCL12(3–68) 激活 ACKR3 的功效显着降低。减弱凝血酶介导的 hPPAEC 屏障功能损伤的效力为：CXCL12α/β、CXCL121、CXCL12-K27A/R41A/R47A > CXCL12-S-S4V、CXCL12-R47E > CXCL122 > CXCL12(3-68)。我们的研究结果表明，CXCR4 激活可减轻凝血酶诱导的肺内皮屏障功能损伤，并表明 CXCL12 的保护作用取决于其 CXCR4 激动剂活性以及不同蛋白质部分与内皮表面硫酸乙酰肝素的相互作用。这些数据可能有助于开发具有改善的药理学特性的化合物，以减少肺循环中凝血酶诱导的血管渗漏。
Recent evidence suggests that chemokine CXCL12, the cognate agonist of chemokine receptors CXCR4 and ACKR3, reduces thrombin-mediated impairment of endothelial barrier function. A detailed characterization of the effects of CXCL12 on thrombin-mediated human lung endothelial hyperpermeability is lacking and structure-function correlations are not available. Furthermore, effects of other CXCR4/ACKR3 ligands on lung endothelial barrier function are unknown. Thus, we tested the effects of a panel of CXCR4/ACKR3 ligands (CXCL12, CXCL11, ubiquitin, AMD3100, TC14012) and compared the CXCR4/ACKR3 activities of CXCL12 variants (CXCL12α/β, CXCL12(3–68), CXCL121, CXCL122, CXCL12-S-S4V, CXCL12-R47E, CXCL12-K27A/R41A/R47A) with their effects on human lung endothelial barrier function in permeability assays. CXCL12α enhanced human primary pulmonary artery endothelial cell (hPPAEC) barrier function, whereas CXCL11, ubiquitin, AMD3100 and TC14012 were ineffective. Pre-treatment of hPPAEC with CXCL12α and ubiquitin reduced thrombin-mediated hyperpermeability. CXCL12α-treatment of hPPAEC after thrombin exposure reduced barrier function impairment by 70% (EC50 0.05–0.5nM), which could be antagonized with AMD3100; ubiquitin (0.03–3μM) was ineffective. In a human lung microvascular endothelial cell line (HULEC5a), CXCL12α and ubiquitin post-treatment attenuated thrombin-induced hyperpermeability to a similar degree. CXCL12(3–68) was inefficient to activate CXCR4 in Presto-Tango β-arrestin2 recruitment assays; CXCL12-S-S4V, CXCL12-R47E and CXCL12-K27A/R41A/R47A showed significantly reduced potencies to activate CXCR4. While the potencies of all proteins in ACKR3 Presto-Tango assays were comparable, the efficacy of CXCL12(3–68) to activate ACKR3 was significantly reduced. The potencies to attenuate thrombin-mediated hPPAEC barrier function impairment were: CXCL12α/β, CXCL121, CXCL12-K27A/R41A/R47A > CXCL12-S-S4V, CXCL12-R47E > CXCL122 > CXCL12(3–68). Our findings indicate that CXCR4 activation attenuates thrombin-induced lung endothelial barrier function impairment and suggest that protective effects of CXCL12 are dictated by its CXCR4 agonist activity and interactions of distinct protein moieties with heparan sulfate on the endothelial surface. These data may facilitate development of compounds with improved pharmacological properties to attenuate thrombin-induced vascular leakage in the pulmonary circulation.
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影响因子：
5.2
作者：
Gur-Cohen S;Kollet O;Graf C;Esmon CT;Ruf W;Lapidot T
通讯作者：
Lapidot T
DOI：
10.1073/pnas.1101133108
发表时间：
2011-10-25
影响因子：
11.1
作者：
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通讯作者：
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作者：
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通讯作者：
Proost, Paul
影响因子：
3.8
作者：
Earle, SA;Proctor, KG;Majetschak, M
通讯作者：
Majetschak, M
影响因子：
8.8
作者：
Baker TA;Romero J;Bach HH 4th;Strom JA;Gamelli RL;Majetschak M
通讯作者：
Majetschak M