Spatial Colocalization and Molecular Crosstalk of Myofibroblastic CAFs and Tumor Cells Shape Lymph Node Metastasis in Oral Squamous Cell Carcinoma

Authors

Ken Furudate
Shuya Kasai
Tadashi Yoshizawa
Yuya Sasaki
Kohei Fujikura
Shintaro Goto
Ryohei Ito
Koki Takagi
Tomoyuki Tanaka
Hiroshi Kijima
Kosei Kubota
Ken Itoh
Wataru Kobayashi
Koichi Takahashi

Publication Date

9-1-2025

Journal

PLoS Genetics

DOI

10.1371/journal.pgen.1011791

PMID

40906645

PMCID

PMC12410789

PubMedCentral® Posted Date

9-4-2025

PubMedCentral® Full Text Version

Post-print

Abstract

Lymph node metastasis (LNM) is a critical prognostic factor for patients with oral squamous cell carcinoma (OSCC). Previous research has implicated the partial epithelial-to-mesenchymal transition of tumor cells and myofibroblastic cancer-associated fibroblasts (myCAFs) in the LNM process. However, the underlying molecular mechanisms remain poorly understood. Here, we conducted a comprehensive molecular analysis integrating original and publicly available OSCC data from bulk genome and transcriptome, single-cell transcriptome, and spatial transcriptome analyses. We found that myCAFs were quantitatively and functionally activated in LNM-positive samples and spatially colocalized with OSCC cells within the invasive tumor front (ITF), providing a niche that may facilitate LNM. Immunohistochemical validation in 90 ITF samples confirmed significantly higher myCAF density in LNM-positive samples than in LNM-negative samples, and this density remained an independent predictor of LNM when adjusted for pathological grade and the pattern of invasion. In LNM-positive samples, myCAFs provided increased extracellular matrix (ECM) signals, upregulating stemness-related genes such as CD44 in OSCC cells. The functional importance of this myCAF-driven ECM-CD44 axis was further supported by our validation analysis of expanded, publicly available spatial transcriptome and experimental in vitro coculture data. We also extracted a spatially resolved, 23-gene signature from the metastatic ITF where OSCC and myCAFs colocalize. This signature predicted LNM status and poor overall survival in patients with OSCC. Our findings provide novel insight into the molecular myCAF/OSCC crosstalk that facilitates LNM and identify potential prognostic biomarkers and therapeutic targets for patients with OSCC.

Keywords

Humans, Mouth Neoplasms, Lymphatic Metastasis, Epithelial-Mesenchymal Transition, Carcinoma, Squamous Cell, Cancer-Associated Fibroblasts, Male, Female, Gene Expression Regulation, Neoplastic, Cell Line, Tumor, Transcriptome, Middle Aged, Squamous Cell Carcinoma of Head and Neck, Extracellular Matrix, Gene Expression Profiling

Published Open-Access

yes

Recommended Citation

Furudate, Ken; Kasai, Shuya; Yoshizawa, Tadashi; et al., "Spatial Colocalization and Molecular Crosstalk of Myofibroblastic CAFs and Tumor Cells Shape Lymph Node Metastasis in Oral Squamous Cell Carcinoma" (2025). Faculty, Staff and Student Publications. 5042.
https://digitalcommons.library.tmc.edu/uthgsbs_docs/5042