胱抑素SN(CST1)和胱抑素SA(CST2)是半胱氨酸蛋白酶抑制剂,可预防过敏原,病毒,和细菌蛋白酶。胱抑素在过敏性鼻炎和慢性鼻窦炎伴鼻息肉(CRSwNP)的背景下过表达;然而,它们在促进2型炎症中的作用仍未得到充分表征.
这项研究的目的是使用整合的多聚组学和鼠暴露模型来探索CRSwNP中胱抑素过表达与2型炎症之间的联系。
在这项机构审查委员会和机构动物护理和使用委员会批准的研究中,我们比较了组织,exosome,通过使用匹配的全外显子组测序,CRSwNP和对照之间的粘液CST1和CST2(每组n=10),转录组,蛋白质组学,翻译后修饰,组织学,功能,和生物信息学分析。在存在或不存在上皮ABCB1a敲低的情况下,对C57/BL6小鼠鼻内给予3.9μg/mL的CST1或PBS持续5至18天。通过使用Quansys多重测定或ELISA定量炎性细胞因子。
在定量的1305种蛋白质中,CST1和CST2是组织中过度表达最多的蛋白酶抑制剂,exosome,和粘液样本;它们位于上皮层。在息肉组织中鉴定了多种翻译后修饰。外泌体CST1和CST2与嗜酸性粒细胞和Lund-Mackay评分密切相关。CST1暴露后,小鼠2型细胞因子分泌和TH2细胞浸润以时间依赖性方式增加,并通过ABCB1a的上皮敲除而消除,上皮细胞因子分泌的调节剂。
CST1是CRSwNP中上皮源性2型炎症的有效上游引发剂。针对CST活性及其相关翻译后修饰的治疗策略值得进一步研究。
Cystatin SN (CST1) and cystatin SA (CST2) are cysteine protease inhibitors that protect against allergen, viral, and bacterial proteases. Cystatins are overexpressed in the setting of allergic rhinitis and chronic rhinosinusitis with nasal polyps (CRSwNP); however, their role in promoting type 2 inflammation remains poorly characterized.
The purpose of this study was to use integrated poly-omics and a murine exposure model to explore the link between cystatin overexpression in CRSwNP and type 2 inflammation.
In this institutional review board- and institutional animal care and use committee-approved study, we compared tissue, exosome, and mucus CST1 and CST2 between CRSwNP and controls (n = 10 per group) by using matched whole exome sequencing, transcriptomic, proteomic, posttranslational modification, histologic, functional, and bioinformatic analyses. C57/BL6 mice were dosed with 3.9 μg/mL of CST1 or PBS intranasally for 5 to 18 days in the presence or absence of epithelial ABCB1a knockdown. Inflammatory cytokines were quantified by using Quansys multiplex assays or ELISAs.
Of the 1305 proteins quantified, CST1 and CST2 were among the most overexpressed protease inhibitors in tissue, exosome, and mucus samples; they were localized to the epithelial layer. Multiple posttranslational modifications were identified in the polyp tissue. Exosomal CST1 and CST2 were strongly and significantly correlated with eosinophils and Lund-Mackay scores. Murine type 2 cytokine secretion and TH2 cell infiltration increased in a time-dependent manner following CST1 exposure and was abrogated by epithelial knockdown of ABCB1a, a regulator of epithelial cytokine secretion.
CST1 is a potent upstream initiator of epithelial-derived type 2 inflammation in CRSwNP. Therapeutic strategies targeting CST activity and its associated posttranslational modifications deserve further interrogation.