Mechanisms Of Npy/Tgfβ Crosstalk-Driven Pni Invasion In Oscc

To investigate the functional interplay between Neuropeptide Y (NPY) signaling and its crosstalk with TGFβ pathways in oral squamous cell carcinoma (OSCC) progression and perineural invasion (PNI). Molecular landscape of NPY and NPY1R was systematically interrogated in OSCC through multiplex IHC, qRT-PCR, and integrative analysis of TCGA (n=502). Functional characterization of NPY1R signaling crosstalk was performed using Cal27 and SCC9 cell lines, employing lentiviral shRNA-mediated knockdown and overexpression systems. Mechanistic studies included phosphoproteomic profiling (ERK/Smad2 activation), transwell invasion assays, and 3D tumorosphere culture in growth factor-reduced matrices. In vivo validation utilized orthotopic tongue xenograft models in NTG mice, with PNI progression monitored via histomorphometric analysis. Multiplex imaging analysis of OSCC specimens revealed significant enrichment of NPY/NPY1R and TβRI within PNI niches, with NPY1R expression levels demonstrating increase in high-grade tumors compared to well-differentiated counterparts. Mechanistically, NPY-NPY1R signaling engaged in bidirectional crosstalk with TGFβ pathways through TβRI transactivation, evidenced increase of ERK phosphorylation and upregulation of Smad2 activation in NPY1R-overexpressing cells. Functional validation demonstrated this axis promoted tumor sphere formation and facilitated tumor cells migration. In orthotopic xenograft models, NPY1R+ tumors exhibited accelerated PNI progression with histopathological confirmation of neural sheath infiltration. The NPY/NPY1R axis collaborates with TGFβ signaling to activate ERK and Smad2 pathways, driving epithelial-neural interaction and enhancing tumor cell motility within perineural niches. This finding nominates NPY1R as a therapeutic target for disrupting tumor-neural crosstalk in OSCC.