Novel molecular mechanisms of immune evasion in hepatocellular carcinoma: NSUN2‐mediated increase of SOAT2 RNA methylation

Abstract Background Hepatocellular carcinoma (HCC) is a deadly malignancy known for its ability to evade immune surveillance. NOP2/Sun RNA methyltransferase family member 2 (NSUN2), an RNA methyltransferase involved in carcinogenesis, has been associated with immune evasion and energy metabolism reprogramming. This study aimed to examine the molecular mechanisms underlying the involvement of NSUN2 in immune evasion and metabolic reprogramming of HCC. Methods Single‐cell transcriptomic sequencing was applied to examine cellular composition changes, particularly immune cell dynamics, in HCC and adjacent normal tissues. Bulk RNA‐seq and proteomics identified key genes and proteins. Methylation sequencing and methylated RNA immunoprecipitation (MeRIP) were carried out to characterize the role of NSUN2 in 5‐methylcytosine (m5C) modification of sterol O‐acyltransferase 2 ( SOAT2 ). Clinical samples from 30 HCC patients were analyzed using reverse transcription‐quantitative polymerase chain reaction and Western blotting. Gene expression was manipulated using CRISPR/Cas9 and lentiviral vectors. In vitro co‐culture models and metabolomics were used to study HCC cell‐T cell interactions, energy metabolism, and immune evasion. Tumor growth in an orthotopic mouse model was monitored by bioluminescence imaging, with subsequent measurements of tumor weight, volume, and immunohistochemical staining. Results Single‐cell transcriptomic analysis identified a marked increase in malignant cells in HCC tissues. Cell communication analysis indicated that tumor cells might promote cancer progression by evading immune clearance. Multi‐omics analyses identified NSUN2 as a key regulator in HCC development. MeRIP confirmed that NSUN2 facilitated the m5C modification of SOAT2 . Analysis of human HCC tissue samples demonstrated pronounced upregulation of NSUN2 and SOAT2 , along with elevated m5C levels in HCC tissues. In vitro experiments uncovered that NSUN2 augmented the reprogramming of energy metabolism and repressed the activity and cytotoxicity of CD8 + T cells, contributing to immune evasion. In vivo studies further substantiated the role of NSUN2 in fostering immune evasion and tumor formation of HCC by modulating the m5C modification of SOAT2 . Conclusions The findings highlight the critical role of NSUN2 in driving HCC progression through the regulation of m5C modification on SOAT2 . These findings present potential molecular markers for HCC diagnosis and therapeutic targets for its treatment.