Spesolimab治疗反常性痤疮小鼠( Ncstn^(flox/flox);K5- Cre)疗效与机制  

Therapeutic effect and mechanism of Spesolimab in the therapy of acne inversa in mice(Ncstn^(flox/flox);K5-Cre)

作　　者：钟奇志 范仕棋 王蓉蓉[1] 张学[1] Zhong Qizhi;Fan Shiqi;Wang Rongrong;Zhang Xue(Institute of Basic Medical Sciences Chinese Academy of Medical Sciences,School of Basic Medicine Peking Union Medical College McKusick-Zhang Center for Genetic Medicine State Key Laboratory for Complex Severe and Rare Diseases,Beijing 100005,China)

机构地区：[1]中国医学科学院基础医学研究所,北京协和医学院基础学院,麦库西克-张孝骞协和遗传医学中心,疑难重症及罕见病国家重点实验室,北京100005

出　　处：《国际遗传学杂志》2024年第3期161-170,共10页International Journal of Genetics

基　　金：国家重点研发计划项目(2022YFC2703900);中国医学科学院医学与健康科技创新工程项目(2021-I2M-1-018);国家自然科学基金(82394420、82394423)。

摘　　要：目的以皮肤K5(Keratin 5)条件性敲除Ncstn(Ncstn^(flox/flox);K5-Cre)反常性痤疮(acne inversa, AI)小鼠模型为研究对象, 采用白介素-36受体抑制剂Spesolimab进行治疗, 为AI寻找新的治疗方法。方法对出生15d和30 d的(Ncstn^(flox/flox);K5-Cre)小鼠及其同窝对照K5-Cre鼠进行单细胞测序并分析(每组3只, 共12只);将出生10d的(Ncstn^(flox/flox);K5-Cre)小鼠随机分成两组, 实验组皮下注射Spesolimab(10 μL/g/2 d), 对照组注射PBS, 小鼠年龄为50 d时取材。使用苏木精-伊红染色法观察小鼠皮肤组织学改变, 利用实时荧光定量PCR和蛋白免疫印迹法检测细胞因子以及炎症因子的相对表达量以评估疗效。结果与PBS组相比, 治疗组小鼠脱毛减少, 皮肤组织角化现象减轻, 同时炎症细胞的浸润也有所减少。利用单细胞测序所检测出在AI小鼠中升高的细胞因子, 在治疗组小鼠中的表达明显降低, 如炎症细胞因子肿瘤坏死因子受体超家族成员9(tumor necrosis factor receptor superfamily member 9, Tnfrs9)(P=0.002)。治疗组各炎症因子蛋白表达水平低于PBS对照组(均P<0.05)。结论 Spesolimab对AI小鼠模型有一定的治疗效果, 可能为AI提供新的治疗思路。Objective A mouse model of cutaneous K5(Keratin 5)conditional knockout Nsctn(Ncstn^(flox/flox);K5-Cre)paradoxical acne inversa(AI)was investigated and treated with interleukin-36(IL-36)receptor inhibitor Spesolimab to find a new therapeutic strategy for AI.Methods Single-cell RNA sequencing of(Ncstn^(flox/flox);K5-Cre)mice and the littermate control K5-Cre mice of 15 and 30 days(3 per group,12 in total);The 10-day-old(Ncstn^(flox/flox);K5-Cre)mice were randomly divided into two groups,and Spesolimab or PBS was subcutaneously injected with a dosage of 10μL/g every 2 days,and then the skin samples were analyzed when the mice were 50 days old.Hematoxylin-Eosin staining was used to observe the histologic changes of mouse skin,and quantitative real-time PCR and western blotting were used to detect the relative expression of cytokines as well as inflammatory factors to assess the therapeutic efficacy.Results Compared with the untreated control group,the treated mice had less skin tissue damage and less inflammatory cell infiltration.The relative expression of cytokines,that had been found to be elevated in AI mice via single-cell RNA sequencing,were significantly decreased in the treated group,such as tumor necrosis factor receptor superfamily member 9(Tnfrs9)(P=0.002).The expression levels of inflammatory cytokines were lower in the treatment group than in the PBS group(all P<0.05).Conclusion Spesolimab shows a promising therapeutic effect on the AI mouse model,which may provide new therapeutic strategies for AI.

关 键 词：反常性痤疮 单细胞测序 IL-36受体抑制剂 Spesolimab 炎症因子 

分 类 号：R758.733[医药卫生—皮肤病学与性病学]