Identification of Regulatory Cell Death-Related Genes during MASH Progression Using Bioinformatics Analysis and Machine Learning Strategies

Zhiqiang Lin¹Weiyi Li^2,3Yuan Xu¹Hangchi Liu¹Yufei Zhang¹Ruifen Li¹Wenqian Zhao¹Youfei Guan^4*Xiaoyan Zhang^1*

• ¹Health Science Center, East China Normal University, Shanghai, China
• ²School of Medicine, Nantong University, Nantong, China
• ³Department of Nephrology,Affiliated Hospital of Nantong University, Nantong, China
• ⁴Advanced Institute for Medical Sciences, Dalian Medical University, Dalian, Liaoning Province, China

Background: Metabolic dysfunction-associated steatohepatitis (MASH) is becoming increasingly prevalent. Regulated cell death (RCD) has emerged as a significant disease phenotype and may act as a marker for liver fibrosis. The present study aimed to elucidate the role of RCD in the progression of MASH.Methods: The gene expression profiles from the GSE130970 and GSE49541 datasets were retrieved from the GEO database for analysis. A total of 101 combinations of 10 machine learning algorithms were employed to screen for characteristic RCD-related differentially expressed genes(DEGs) that reflect the progression of NASH. GO and KEGG analyses were conducted to explore the enrichment pathways and functions of the RCD-related DEGs. we performed cell classification analysis to investigate immune cell infiltration. Consensus cluster analysis was performed to identify MASH subtypes associated with RCD. The GEPIA2 tool was utilized to display the expression levels of key genes in liver cancer tissues compared to adjacent tissues. The DGIdb database was employed to screen for potential therapeutic drugs targeting the RCD-related DEGs. Lastly, we established mouse liver fibrosis models induced by MCD diet or CCl4 treatment, and further validated the expression of characteristic genes through q-PCR.Results: This study discovered a total of 11 characteristic DEGs. The expression levels of RCD-related DEGs were higher in advanced MASH and significantly correlated with the infiltration of immune cells.MASH can be classified into two subtypes, cluster 1 and cluster 2, based on these DEGs. Compared with cluster 2, cluster 1 has highly expressed RCD-related DEGs and is closely related to inflammation and immune regulation. Furthermore, we observed elevated expression levels of certain characteristic genes in the liver tissue of patients with hepatocellular carcinoma. Subsequently, we used the DGIDB database to predict 30 drugs that may interact with RCD-related DEGs. Finally, we evaluated the expression of these 11 characteristic genes in liver tissue of mice with MCD or CCl4-induced fibrosis, and the results suggested that these genes may be involved in the development of fibrosis.Conclusion: Our study sheds light on the fact that RCD contribute to the progression of MASH, high lighting potential therapeutic targets for treating this disease.