Comparative analysis of patient-derived organoids among oral squamous cell carcinoma and mucosa identifies the distinct features.

OBJECTIVE

This study aimed to establish matched patient-derived organoids from oral squamous cell carcinoma (PDO-T) and adjacent normal mucosa (PDO-N), characterize their distinct features, and evaluate their impact on drug response.

DESIGN

PDOs were generated from primary OSCC tissues and matched adjacent normal mucosa obtained from 18 patients. Morphological and histological characteristics were assessed using brightfield microscopy and phalloidin staining. Immunohistochemistry was performed to evaluate differential marker expression. Drug response assays were conducted in paired PDO-T and PDO-N to assess chemotherapeutic sensitivity.

RESULTS

We successfully established 5 PDO-T and 15 PDO-N lines, with PDO-N exhibiting a higher success rate (∼80 %). Morphological analysis showed that PDO-N formed spherical structures with a central cavity and polarity, while PDO-T displayed irregular structures with heterogeneous cell arrangements. Immunohistochemistry revealed broad distribution of Ki-67, p63 and YAP/TAZ in PDO-T, contrasting with their basal layer restriction in PDO-N. p53 was strongly positive in PDO-T. CK5 and CK13 exhibited a stratified pattern in PDO-N. Importantly, drug testing in paired PDO-T and PDO-N revealed divergent responses, with PDO-N generally more resistant and able to maintain morphological integrity under high-dose treatment.

CONCLUSION

Our comparative analysis delineates distinct features between PDO-T and PDO-N in morphology, biomarker expression, and drug response. These findings provide an intuitive framework for their identification, thereby improving the accuracy of drug screening and enhancing the translational value of PDOs.