Exploring the role of HOXB13, a regulator gene, in fatty acid metabolism and gastric epithelial cell intestinalization during Helicobacter pylori Infection: insights from single- cell RNA sequencing analysis

Abstract

Abstract Background To systematically identify the changes in epithelial cell types during the gastrointestinal epithelial transformation, further explore the dynamic changes of key transcription factors and metabolic pathways during the gastrointestinal metaplasia process, and investigate the specific role of Helicobacter pylori (H. pylori) in this process. Methods Based on an in-depth analysis of single cell sequencing data, we identified specific transcription factors involved in the gastric epithelial intestinalization process using SCENIC analysis and validated them through immunohistochemistry. Based on these findings, we further investigated the differentiation trajectory of gastric epithelial cells during metaplasia and examined changes in metabolic pathways throughout this process. Finally, we studied the impact of H. pylori infection on these specific transcription factors using PCR analysis and Western blotting. Results: During the process of intestinal metaplasia, we have confirmed that the proportion of pit mucous cells gradually decreases while the number of intestinal metaplasia cells increases. Through SCENIC analysis, we have identified HOXB13, HOXA10, and ONECUT2 as potential characteristic transcription factors in intestinal metaplasia cells. Additionally, immunohistochemistry validation has shown a significant increase in HOXB13 expression in intestinal metaplasia tissues compared to non-atrophic gastritis tissues. In the non-atrophic gastritis group, the expression of gastric tissue infected with H. pylori was higher than that of the non-infected group. Trajectory analysis has revealed that pathways related to lipid metabolism play important roles during the process of intestinal metaplasia. Furthermore, cell experiments have confirmed a significant upregulation of HOXB13 after H. pylori infection. Conclusion This single cell RNA sequencing analysis establishes a solid foundation for investigating the cellular types, key transcription factors, and metabolic pathways associated with gastrointestinal epithelial cell differentiation. Our findings indicate that HOXB13 plays a critical role as a transcription factor in gastrointestinal epithelial metaplasia. Additionally, we have observed that H. pylori infection enhances the expression of HOXB13, which subsequently impacts fatty acid metabolism in normal gastric mucosa through downstream target genes CEL and GPD1. As a result, HOXB13 emerges as a promising therapeutic target to address H. pylori-induced intestinal metaplasia and offers potential insights into the development of treatment strategies for this condition.