Omics insights into MYBL2-promoted cancer stem-like cells driving ferroptosis resistance in hepatocellular carcinoma.

[INTRODUCTION] Cancer stem cells are critically involved in the initiation and progression of hepatocellular carcinoma (HCC). Understanding their biological properties and underlying mechanisms is essential for the development of effective targeted therapies.

[OBJECTIVES] This study aims to identify the key stem-like cell subtype and elucidate the molecular mechanisms underlying its tumor-promoting effects in HCC.

[METHODS] Single-nucleus RNA sequencing (snRNA-seq) of the c-Myc/Cre transgenic HCC mouse model and single-cell RNA sequencing (scRNA-seq) from HCC patients were analyzed to identify key tumorigenic subpopulation with stemness properties. Integrated analysis of snRNA/scRNA-seq, bulk RNA sequencing, and spatial transcriptomics data elucidated the biological features, molecular signatures, and intercellular interactions of the identified subpopulation. In vitro experiments were performed to validate the functional role of the key regulatory factor. A risk score model based on relevant gene signatures was subsequently developed to evaluate the clinical significance.

[RESULTS] We mapped the hepatic transcriptomic landscape of the c-Myc/Cre transgenic HCC mouse model using snRNA-seq and identified cancer stem-like cells (CSLCs) with pronounced stemness properties as key drivers of HCC progression. scRNA-seq analysis of HCC patient tissues reveals that patient-derived CSLCs exhibited cross-species similarities with mouse-derived CSLCs and were strongly associated with tumor metastasis and poor prognosis. CSLCs actively suppressed lipid peroxidation and exhibited significant ferroptosis resistance, facilitating their survival within the tumor microenvironment. Mechanistically, MYB Proto-Oncogene Like 2 (MYBL2) was specifically activated in CSLCs, and MYBL2 knockdown effectively impaired cancer stemness and reduced ferroptosis resistance. Notably, our newly developed six-gene CSLC signature (CSLC.Sig) model achieved excellent prognostic prediction in HCC. Furthermore, strong interaction and spatial "neighborhood" localization were observed between CSLCs and activated hepatic stellate cells, indicating a synergistic pro-tumorigenic niche.

[CONCLUSION] Our study identifies MYBL2-driven CSLCs as critical players in maintaining cancer stemness and ferroptosis resistance, providing potential therapeutic targets to disrupt CSLCs-mediated tumor progression in HCC.