A Single-Cell and Spatial Transcriptomic Analysis of Conventional Ameloblastoma.

OBJECTIVE

To understand the cellular and spatial composition of conventional ameloblastoma and its alteration after marsupialization treatment.

MATERIALS AND

METHODS

We applied single-cell RNA sequencing and spatial transcriptomics on pre- and post-marsupialization conventional ameloblastoma slices, followed by cluster, trajectory, network analysis, and immunofluorescence validation.

RESULTS

We found a continuous cell trajectory from pre-marsupialization epithelium-like cell types to post-marsupialization mesenchyme-like cell types. Comparison with our recently established human embryonic dental cell atlas revealed that epithelium-like cells resembled the most primitive outer enamel progenitors, whereas mesenchyme-like cells did not resemble any embryonic mesenchymal cells, representing a differentiated and abnormal state. Transcription factors DLX5 and TWIST1, together with their target genes, showed profound alterations that paralleled the epithelium-to-mesenchyme transition (EMT). We verified the existence of EMT by double immunofluorescence of two EMT markers FN1 and VIM, and verified the specific expression of KRT15 in pre-marsupialization tissues. Lastly, mesenchyme-derived proteins MMP13 and THBS2 exert feedback regulatory effects on EMT.

CONCLUSIONS

We constructed the cellular and spatial landscape of conventional ameloblastoma before and after marsupialization. Our findings indicate that the epithelial-mesenchymal transition (EMT) process, potentially regulated by DLX5 and MMP13, contributes to the treatment effect of marsupialization.