A novel MRGPRX2-targeting antagonistic DNA aptamer inhibits histamine release and prevents mast cell-mediated anaphylaxis
A novel MRGPRX2-targeting antagonistic DNA aptamer inhibits histamine release and prevents mast cell-mediated anaphylaxis
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DOI：
10.1016/j.ejphar.2020.173104
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发表时间：
2020-07-05
影响因子：
5
通讯作者：
Mogi, Masaki
中科院分区：
文献类型：
--
作者：
Suzuki, Yasuyuki;Liu, Shuang;Mogi, Masaki
全麻期间的过敏反应是麻醉医师面临的重大临床挑战。大约50%的围手术期过敏反应病例缺乏特异性IgE抗体的存在。人类肌浆蛋白相关g蛋白偶联受体X2 （MRGPRX2）及其小鼠同源物肌浆蛋白相关g蛋白偶联受体B2 （Mrgprb2）是非ige依赖性组胺释放的关键受体。麻醉剂如罗库溴铵和阿曲库铵通过Mrgprb2途径激活组胺释放引起围手术期过敏反应。我们假设靶向MRGPRX2的拮抗DNA适体可以预防围手术期过敏反应。通过使用我们改良的配体系统进化指数富集（SELEX）方法，选择特异性结合MRGPRX2的DNA适体。我们的SELEX工艺使用小麦无生殖细胞系统合成的mrgprx2蛋白脂质体作为模板。选择的适体抑制mrgprx2激活肥大细胞和移植该细胞系的过敏反应大鼠模型中组胺释放的活性进行了检测。我们的筛选过程确定了适体- x35的序列为5‘-ATGACCATGA CCCTCCACACTGTAGGCACCACGGGTCCCTGGCAGTTAAAAGTACGTTGTCAGACTGTGGCAGGGAAACA-3’。适配体- x35的硅二维模型显示了一个小环和长茎的结构。Aptamer-X35抑制肥大细胞组胺释放70%。皮下注射30 nmol适配体- x35对大鼠过敏反应有抑制作用。本研究表明，改良SELEX方法选择的适体x35通过抑制MRGPRX2途径减少组胺释放。总之，我们的研究结果确立了适体- x35作为围手术期过敏反应的潜在治疗候选药物。
Anaphylaxis during general anaesthesia is a significant clinical challenge for anaesthesiologists. Approximately 50% of perioperative anaphylaxis cases lack the presence of specific IgE antibodies. Mas-related G-protein coupled receptor X2 (MRGPRX2) in humans and its mouse orthologue Mas-related G-protein coupled receptor B2 (Mrgprb2) are crucial receptors in non-IgE-dependent histamine release. Anaesthetics such as rocuronium and atracurium cause perioperative anaphylaxis by activating histamine release via the Mrgprb2 pathway. We hypothesized that antagonistic DNA aptamers that target MRGPRX2 can prevent perioperative anaphylaxis. Selection of a DNA aptamer that specifically binds MRGPRX2 was achieved by using our modified Systematic Evolution of Ligands by Exponential enrichment (SELEX) approach. Our SELEX process used MRGPRX2-proteoliposomes synthesised by a wheat germ cell-free system as templates. The activity of the selected aptamer to inhibit histamine release from MRGPRX2-activated mast cells and in an anaphylaxis rat model transplanted with this cell line was examined. Our selection process identified aptamer-X35 with the sequence 5'-ATGACCATGA CCCTCCACACTGTAGGCACCACGGGTCCCTGGCAGTTAAAAGTACGTTGTCAGACTGTGGCAGGGAAACA-3'. In silico 2D modelling of aptamer-X35 revealed a structure with a small loop and a long stem. Aptamer-X35 inhibited histamine release from mast cells by 70%. Subcutaneous injection of 30 nmol of aptamer-X35 inhibited the anaphylactic reaction in the rat anaphylaxis model.This study demonstrated that aptamer-X35 selected by the modified SELEX approach reduced histamine release by inhibiting the MRGPRX2 pathway. Overall, our findings establish aptamer-X35 as a potential therapeutic candidate against perioperative anaphylaxis.