Amelioration of Compound 48/80-Mediated Itch and LL-37-Induced Inflammation by a Single-Stranded Oligonucleotide.
Amelioration of Compound 48/80-Mediated Itch and LL-37-Induced Inflammation by a Single-Stranded Oligonucleotide.
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单链寡核苷酸对化合物48/80介导的瘙痒和LL-37诱导的炎症的改善。
DOI：
10.3389/fimmu.2020.559589
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发表时间：
2020
影响因子：
7.3
通讯作者：
Spetz AL
中科院分区：
文献类型：
--
作者：
Dondalska A;Rönnberg E;Ma H;Pålsson SA;Magnusdottir E;Gao T;Adam L;Lerner EA;Nilsson G;Lagerström M;Spetz AL
许多炎症性皮肤疾病显示瘙痒的高患病率。mass相关的G蛋白偶联受体X2 （MRGPRX2）已被证明通过诱导非ige介导的肥大细胞脱颗粒和释放内源性瘙痒诱导剂来调节瘙痒。各种统称为碱性分泌物的物质，包括炎症肽和某些药物，可触发MRGPRX2，从而诱发以组胺和蛋白酶释放为特征的假过敏反应，并伴有炎症反应。在这里，我们研究了免疫调节单链寡核苷酸（ssON）调节不依赖ige的肥大细胞脱颗粒的能力，更具体地说，它在体外和体内抑制基本分泌物化合物48/80 （C48/80）和LL-37的能力。我们通过测量人肥大细胞系（LAD2）的脱颗粒和MRGPRX2转染的HEK293细胞中的钙内流，研究了ssON对MRGPRX2体外激活的影响。为了确定ssON对瘙痒的影响，我们在已建立的小鼠模型中进行了行为学研究，并收集了皮肤活检进行组织学分析。此外，我们利用酒渣鼻小鼠模型和RT-qPCR研究了ssON对ll -37诱导炎症的影响。我们发现，在MRGPRX2转染的HEK293细胞中，由抗菌肽LL-37和碱性分泌剂C48/80诱导的肥大细胞脱颗粒和钙内流均被ssON以剂量依赖性的方式有效抑制。此外，在两种小鼠模型中，ssON显示了抑制LL-37和C48/80激活的能力。我们发现，小鼠皮内注射ssON能够以剂量依赖的方式阻断C48/80诱导的瘙痒。组织学染色显示，松对C48/80处理的小鼠皮肤急性肥大细胞脱颗粒有抑制作用。最后，我们在酒渣鼻小鼠模型中表明，ssON治疗可以改善ll -37诱导的炎症。由于需要针对非ige介导的肥大细胞激活的新疗法，因此ssON可作为解决某些皮肤病瘙痒和炎症的潜在候选药物。
Numerous inflammatory skin disorders display a high prevalence of itch. The Mas-related G protein coupled receptor X2 (MRGPRX2) has been shown to modulate itch by inducing non-IgE-mediated mast cell degranulation and the release of endogenous inducers of pruritus. Various substances collectively known as basic secretagogues, which include inflammatory peptides and certain drugs, can trigger MRGPRX2 and thereby induce pseudo-allergic reactions characterized by histamine and protease release as well as inflammation. Here, we investigated the capacity of an immunomodulatory single-stranded oligonucleotide (ssON) to modulate IgE-independent mast cell degranulation and, more specifically, its ability to inhibit the basic secretagogues compound 48/80 (C48/80)-and LL-37 in vitro and in vivo. We examined the effect of ssON on MRGPRX2 activation in vitro by measuring degranulation in a human mast cell line (LAD2) and calcium influx in MRGPRX2-transfected HEK293 cells. To determine the effect of ssON on itch, we performed behavioral studies in established mouse models and collected skin biopsies for histological analysis. Additionally, with the use of a rosacea mouse model and RT-qPCR, we investigated the effect on ssON on LL-37-induced inflammation. We reveal that both mast cell degranulation and calcium influx in MRGPRX2 transfected HEK293 cells, induced by the antimicrobial peptide LL-37 and the basic secretagogue C48/80, are effectively inhibited by ssON in a dose-dependent manner. Further, ssON demonstrates a capability to inhibit LL-37 and C48/80 activation in vivo in two mouse models. We show that intradermal injection of ssON in mice is able to block itch induced via C48/80 in a dose-dependent manner. Histological staining revealed that ssON inhibits acute mast cell degranulation in murine skin treated with C48/80. Lastly, we show that ssON treatment ameliorates LL-37-induced inflammation in a rosacea mouse model. Since there is a need for new therapeutics targeting non-IgE-mediated activation of mast cells, ssON could be used as a prospective drug candidate to resolve itch and inflammation in certain dermatoses.