Novel peptide nanoparticle-biased antagonist of CCR3 blocks eosinophil recruitment and airway hyperresponsiveness.
Novel peptide nanoparticle-biased antagonist of CCR3 blocks eosinophil recruitment and airway hyperresponsiveness.
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DOI：
10.1016/j.jaci.2018.05.003
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发表时间：
2019-03
期刊：
影响因子：
--
通讯作者：
Ackerman SJ
中科院分区：
文献类型：
--
作者：
Grozdanovic M;Laffey KG;Abdelkarim H;Hitchinson B;Harijith A;Moon HG;Park GY;Rousslang LK;Masterson JC;Furuta GT;Tarasova NI;Gaponenko V;Ackerman SJ
通过 CCR3 的趋化因子信号传导是嗜酸性粒细胞募集到与过敏性炎症和哮喘相关的组织中的关键调节途径。迄今为止，没有一种 CCR3 拮抗剂在临床试验中显示出疗效。原因之一可能是它们的无偏抑制模式阻止受体内化，从而导致药物耐受。我们试图开发一种新型肽纳米颗粒 CCR3 抑制剂 (R321)，其具有偏向的抑制模式，可阻断 G 蛋白信号传导，但能够或促进受体内化。通过动态光散射和 NMR 分析 R321 肽自组装成纳米颗粒以及肽与 CCR3 的结合。使用流式细胞术、共聚焦显微镜和蛋白质印迹分析在 CCR3+ 嗜酸性粒细胞系和血液嗜酸性粒细胞中体外评估 CCR3 信号传导的抑制活性。使用三重过敏原小鼠哮喘模型评估 R321 的体内作用。 R321 自组装成纳米颗粒并直接与 CCR3 结合，从而改变受体功能。嗜酸粒细胞趋化因子诱导的血液嗜酸性粒细胞趋化作用的 IC50 值在低纳摩尔范围内。 R321 仅抑制 ERK1/2 激活的早期阶段，而不抑制通常与 β-arrestin 募集和受体内吞作用相关的晚期阶段，从而促进 CCR3 内化和降解。在体内，R321 可有效阻止嗜酸性粒细胞募集到肺部和气道，并防止小鼠嗜酸性粒细胞哮喘模型中的气道高反应性。 R321 是 CCR3 信号级联的有效选择性拮抗剂。通过偏向的拮抗模式进行抑制可能通过避免药物耐受性的形成而具有重要的治疗前景。
Chemokine signaling through CCR3 is a key regulatory pathway for eosinophil recruitment into tissues associated with allergic inflammation and asthma. To date, none of the CCR3 antagonists have shown efficacy in clinical trials. One reason may be their unbiased mode of inhibition that prevents receptor internalization, leading to drug tolerance. We sought to develop a novel peptide nanoparticle CCR3 inhibitor (R321) with a biased mode of inhibition that would block G-protein signaling, but enable or promote receptor internalization. Self-assembly of R321 peptide into nanoparticles and peptide binding to CCR3 were analyzed by dynamic light scattering and NMR. Inhibitory activity on CCR3 signaling was assessed in vitro using flow cytometry, confocal microscopy, and western blot analysis in a CCR3+ eosinophil cell line and blood eosinophils. In vivo effects of R321 were assessed using a triple allergen mouse asthma model. R321 self-assembles into nanoparticles and binds directly to CCR3, altering receptor function. IC50 values for eotaxin-induced chemotaxis of blood eosinophils are in the low nanomolar range. R321 inhibits only the early phase of ERK1/2 activation and not the late phase generally associated with β-arrestin recruitment and receptor endocytosis, promoting CCR3 internalization and degradation. In vivo, R321 effectively blocks eosinophil recruitment into the lungs and airways and prevents airway hyperresponsiveness in a mouse eosinophilic asthma model. R321 is a potent and selective antagonist of the CCR3 signaling cascade. Inhibition through a biased mode of antagonism may hold significant therapeutic promise by eluding the formation of drug tolerance.
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DOI：
10.3390/molecules18067103
发表时间：
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期刊：
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影响因子：
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