A patient-derived organoid model identifies TP53-dependent gemcitabine sensitivity in spinal chordoma

Chordoma is a rare malignant bone tumor with limited effective systemic treatment options. Conventional chemotherapy generally shows minimal benefit, highlighting the need for predictive preclinical models to explore therapeutic vulnerabilities. Patient-derived organoids (PDOs) have emerged as a promising three-dimensional culture system that preserves tumor architecture and molecular features while enabling functional testing. However, mechanistic studies linking pathway activity to drug response in chordoma PDOs remain limited.

Fresh surgical specimens from five patients with primary spinal chordoma were used to attempt generation of three-dimensional PDO cultures, of which primary three-dimensional cultures were successfully established. Organoids were characterized by histology, immunohistochemistry, and quantitative PCR analysis of chordoma-associated markers. Functional drug screening was performed using a panel of clinically relevant agents in a representative PDO model, followed by dose-response testing of gemcitabine. The role of TP53 in drug response was examined using small interfering RNA-mediated knockdown, with assessment of cell viability, Ki-67 expression, and DNA-damage response-related proteins. Statistical analyses were performed using one-way analysis of variance, with p < 0.05 considered statistically significant.

The established PDOs recapitulated the histopathological and molecular characteristics of their matched primary tumors and maintained stable growth across six passages. Among the screened agents, gemcitabine showed the strongest growth-inhibitory effect in PDO-based functional assays. Dose-response experiments confirmed significant gemcitabine-induced growth suppression. Importantly, TP53 knockdown markedly attenuated gemcitabine-induced cytotoxicity, increased proliferative activity, and reduced activation of DNA-damage response signaling, indicating a TP53-dependent vulnerability.

This study establishes a spinal chordoma PDO platform for functional precision oncology. Our findings identify a TP53-dependent DNA-damage vulnerability engaged by gemcitabine in patient-derived three-dimensional models, supporting biomarker-informed hypothesis generation rather than routine chemotherapy in unselected patients. This PDO-based approach provides a translational framework for exploring pathway-defined therapeutic susceptibilities in rare tumors such as chordoma.