抑制IL-4和IL-13是诱导过敏性气道疾病的致病反应的关键细胞因子。目前,靶向IL-4Rα的单克隆抗体皮下给药以治疗嗜酸性粒细胞性鼻窦炎和过敏性哮喘。然而,这些治疗有几个缺点。为了解决这些问题,我们开发了一种新型IL-4Rα靶向纳米抗体,设计用于过敏性气道疾病的非侵入性递送至局部炎症部位.H5,通过核糖体展示从合成纳米抗体库应用筛选选择,使用AlphaFold2和GROMACS进行二聚化和计算机亲和力成熟,导致其结合亲和力显着增强。H5有效控制炎症标志物如MUC5AC,CCL26和FOXJ1在人鼻上皮细胞(HNEC)中通过抑制IL-4和IL-13信号传导。H5的二价形式在易于接近的细胞中显示出功效,如多纤毛细胞,而单价变体靶向难以触及的细胞,如HNECs的基底细胞。总之,我们开发了一种纳米抗体,可以通过鼻内给药有效抑制HNECs中的炎症信号,显示作为非侵入性鼻炎治疗的希望。
Inhibiting IL-4 and IL-13 are critical cytokines that induce the pathogenic responses of allergic airway diseases. Currently, monoclonal antibodies targeting IL-4Rα are administered subcutaneously to treat eosinophilic rhinosinusitis and allergic asthma. However, these treatments have several drawbacks. To address these issues, we have developed a novel IL-4Rα-targeting nanobody designed for non-invasive delivery to local inflammatory sites in allergic airway diseases. H5, selected via the ribosomal display applied screening from synthetic nanobody library, underwent dimerization and in-silico affinity maturation using AlphaFold2 and GROMACS resulting in a substantial/dramatic enhancement of its binding affinity. H5 effectively controlled inflammatory markers such as MUC5AC, CCL26, and FOXJ1 in human nasal epithelial cells (HNECs) by inhibiting IL-4 and IL-13 signaling. The bivalent form of H5 showed efficacy in easily accessible cells, such as multi-ciliated cells, while the monovalent variant targeted hard-to-reach cells, such as basal cells of HNECs. In summary, we developed a nanobody that could effectively inhibit inflammatory signaling in HNECs via intranasal administration, showing promise as a non-invasive rhinitis treatment.