Overview

Digestive system cancers remain a significant clinical challenge. By using patient-derived cancer organoids and applying drug sensitivity and organoid assembly assays, we tested new therapeutic protocols developed in our laboratory, comparing them to standard chemotherapy. The organoid model allowed us to monitor how different parameters of the 3D culture respond to the used protocols.

Introduction

Pancreatic and colon cancers are among the most common causes of death in Polish and EU neoplasia patients. Improvement of patient outcomes in these diseases requires personalized therapeutic approaches and a larger choice of treatment protocols. The broad aim of this project is to develop and test new experimental therapeutic protocols using patient-derived cancer organoids.

Methods

>Patient-derived colon and pancreatic cancer organoids >Organoid Drug Sensitivity Assay >Organoid Assembly Assay based on Spinning Disc Microscopy >ImageJ with TrackMate and GraphPad Prism software used for analysis

Results

>The efficiency of experimental therapeutic protocols matched or surpassed standard chemotherapeutic protocols in killing cancer organoids characterized by either TP53 mutations, KRAS mutations, or MYC activation. >Organoid assembly assay allowed to distinguish effects on cell migration in a 3D environment and formation of organoids in time. The used drug combinations decreased only the assembly, while sgRNA-mediated mutant p53 knockdown affected the migration.

Conclusion

>Heterogeneous colon and pancreatic cancer organoid models confirm the high efficacy of novel pre-clinical targeted drug combinations earlier developed using 2D cancer cell line cultures. >Organoid assembly assay allows us to analyze and distinguish how cell migration and cell group formation towards organoids may be differentially affected by anti-cancer drug treatment and direct oncogene targeting.