M. Lin1, K. Hirai1, M. Choi1, D. Tzimas1, J. C. Bucobo1, J. Buscaglia1, G. V. Georgakis1, A. Sasson1, M. Gao1, J. Kim1 1Stony Brook University Medical Center,Stony Brook, NY, USA
Introduction: Comprehensive testing of drugs for advanced gastric cancer is limited by the lack of models that recapitulate human disease. Patient-derived, 3-dimensional organoids have been developed and may accurately model in vivo disease. Diagnostic upper endoscopy is a potential avenue for gastric cancer tissue sampling and organoid creation. Here, we report the first successful creation of organoids from endoscopic biopsy in a patient with gastric adenocarcinoma.
Methods: With IRB approval and written informed consent, upper endoscopy was performed on a patient with large gastric adenocarcinoma. Standard biopsy forceps were used to obtain tissue from three different tumor locations to create three sets of gastric organoids using a modified technique. Biopsy tissues were placed in prepared organoid medium, then washed and isolated to preserve glandular architecture. Gastric glands were collected and plated in 24-well plates. To confirm gastric origin, we performed immunofluorescent staining for LGR5 and TROY protein markers. To assess potential tumor heterogeneity within this patient’s tumor, we also created gastric cancer organoids from surgical resection tissues and obtained whole tumor lysates. We extracted DNA from all organoids and tissues and performed low-coverage, whole genome sequencing.
Results: We successfully obtained upper endoscopy gastric cancer biopsy tissues for creation of organoids. Immediately following resection of the primary tumor, we also obtained tissues for organoid creation and genomic testing. For all organoids, we observed an initial coalescence of gastric glandular tissues into cystic structures. Some cysts developed into spheroids, whereas others remained cystic. Budding structures formed on the periphery of the spheroids after 5-7 days in culture. Gastric origin of the organoids was confirmed based on positive LGR5 and TROY stains. Comparison of whole genome sequencing between endoscopic derived and surgical organoids and whole tumor DNA assessed the tumor homogeneity.
Conclusion: We report the first successful creation of organoids from endoscopic biopsy of gastric adenocarcinoma. The organoids appear to accurately portray human disease and have tremendous potential to be used for personalized drug testing.
