# Spatial Proximity of Cancer-Associated Fibroblasts to Tumor and Osteoclasts Suggests a Coordinating Role in OSCC-Induced Bone Invasion: A Preliminary Study

## TL;DR

This study explores how cancer-associated fibroblasts may help oral cancer invade jawbone by acting as a bridge between tumor cells and bone-resorbing cells.

## Contribution

The study reveals a novel spatial coordination role of CAFs in OSCC-induced bone invasion using AI-assisted histology.

## Key findings

- CAFs are clustered near tumor cells and osteoclasts within 50 μm at the tumor–bone interface.
- AI-assisted image analysis revealed consistent CAF distribution patterns across OSCC specimens.
- CAFs may facilitate signaling between tumor cells and osteoclasts to promote bone resorption.

## Abstract

Background: Jawbone invasion is a common and prognostically unfavorable feature of oral squamous cell carcinoma (OSCC). Although cancer-associated fibroblasts (CAFs) are recognized for their role in tumor progression, their spatial dynamics at the tumor–bone interface remain poorly understood. Methods: We analyzed 14 OSCC specimens with confirmed jawbone invasion using histopathological and immunohistochemical techniques. Digital pathology combined with AI-assisted image analysis was employed to quantify and visualize the spatial distribution of OSCC cells (RANKL-positive), CAFs (α-SMA and FAP-positive), and osteoclasts (cathepsin K-positive) within defined regions of interest at the tumor–bone invasive front. Results: A consistent laminar stromal region enriched in CAFs was observed between the tumor nests and jawbone. CAFs were spatially clustered near OSCC cells and osteoclasts, with 81% and 74% residing within 50 μm, respectively. On average, 11.4 CAFs were present per OSCC cell and 23.2 per osteoclast. These spatial proximities were largely preserved irrespective of stromal thickness, suggesting active bidirectional cellular interactions. Conclusions: Our findings demonstrate that CAFs are strategically positioned to facilitate intercellular signaling between tumor cells and osteoclasts, potentially coordinating OSCC proliferation and bone resorption. This study highlights the utility of AI-assisted spatial histology in unraveling tumor microenvironmental dynamics and proposes CAFs as potential therapeutic targets in OSCC-induced osteolytic invasion.