Super-enhancer-driven TCF4 orchestrates neuroblastoma metastasis by sphingolipid-dependent membrane remodeling and ITGB1-FAK activation

Background: Metastasis remains a critical determinant of survival in neuroblastoma (NB), yet the role of transcriptional dysregulation, particularly super-enhancer (SE)-mediated transcriptional control in this process has not been elucidated. The aim of this study is to identify the SE-driven transcription factors involved in the metastasis of NB and potential targeted drugs. Methods: The metastatic SE-driven Transcription Factor 4 (TCF4) was screened and identified by integrating bioinformatic analyses of H3K27ac ChIP-seq and scRNA-seq. The effect of TCF4 on NB cell metastasis was evaluated through in vivo and in vitro functional experiments. The molecular mechanism of TCF4 was investigated by the study of targeted CUT&Tag and transcriptome sequencing. Results: TCF4 is associated with poor prognosis in patients and significantly promotes the metastasis ability of NB cells both in vivo and in vitro. Mechanistically, TCF4 transcriptionally activates SPTLC1, a pivotal enzyme in sphingolipid biosynthesis, to promote ganglioside GM3 synthesis. GM3 orchestrates membrane architecture remodeling, thereby modulating ITGB1 membrane localization and activation, which subsequently potentiates FAK signaling. Notably, we demonstrate that the HDAC6 inhibitor ACY-1215 suppressed NB malignancy by destabilizing TCF4 protein. Conclusions: Our findings indicate that SE-driven TCF4 can orchestrate metastatic transcriptional networks to maintain NB malignancy and propose ACY-1215 as a translational therapeutic candidate for clinical intervention.