Hp-LPS通过USP13介导VCP去泛素化促进胃黏膜解痉多肽化生的机制研究

幽门螺杆菌（Hp）在胃黏膜解痉多肽化生（SPEM）发生过程中发挥重要作用，但其作用机制仍不明确。前期SPEM模型样本和Hp脂多糖（Hp-LPS）处理胃上皮细胞样本的RNA-seq数据提示泛素特异性蛋白酶13（USP13）可能参与SPEM发生。Co-IP后的质谱分析和预实验结果初步证实USP13可与VCP结合并促进其表达，且VCP的高表达促进SPEM的发生。RNA-seq数据和本课题组已发表文献提示自噬激活可能与SPEM发生相关。由此我们提出“Hp-LPS通过USP13调节VCP翻译后修饰，继而活化自噬通路参与SPEM发生”的科学假说。本课题拟通过细胞，3D-类器官和基因敲除小鼠模型探究USP13/VCP/自噬在Hp-LPS致SPEM中的作用，拟通过GST pull-down，体外去泛素化实验深入探讨USP13对VCP的去泛素化调控机制，为Hp所致SPEM甚至是胃癌提供预防和干预的可能靶点。

Helicobacter pylori (Hp) plays an important role in the occurrence of spasmolytic polypeptide-expressing metaplasia (SPEM), but the mechanism remains unclear. The Ubiquitin specific proteasome 13 (USP13), which may be related to SPEM induced by Helicobacter pylori lipopolysaccharide (Hp-LPS), was screened out according to the conjoint analysis of RNA sequencing of SPEM samples and Hp-LPS treated gastric epithelial cell samples. Combined with the previous results, it was preliminarily proved that USP13 promotes the occurrence of SPEM induced by Hp-LPS. Mass spectrometry analysis after co-immunoprecipitation (Co-IP) and previous experiment results revealed that USP13 could bind to VCP and promote VCP expression to participate in the occurrence of SPEM. The KEGG analysis of RNA-seq showed that autophagy pathway, which was proved activated in gastric cancer according to our previously published research, was enriched in the occurrence of SPEM. Therefore, we hypothesized that Hp-LPS promotes USP13 to regulate VCP in a post-translational level, and then activates the autophagy pathway to participate in the occurrence of SPEM. The current study will deeply explore the role of USP13/VCP/autophagy in the process of SPEM induced by Hp-LPS, All the experiments will be conducted in cells, 3D-organoids and gene knock out animal models. Besides, we discuss the deubiquitinating regulation of USP13 on VCP through GST pull-down and in vitro deubiquitination experiments. In conclusion, all these efforts will provide possible targets for the prevention and intervention of SPEM and even gastric cancer caused by Hp.