OPTIMIZING DEVELOPMENT OF ENDOSCOPIC-DERIVED GASTRIC CANCER ORGANOIDS FOR PERSONALIZED MEDICINE
Miranda Lin*1, Mei Gao1, Yachao Yang1, Wesam M. Frandah2, Moamen Gabr2, Houssam E. Mardini2, Joseph Kim1
1Surgery, University of Kentucky, Lexington, KY; 2Gastroenterology, University of Kentucky, Lexington, KY
Patient-derived organoids (PDOs) are 3D models of human development that have been created for gastric cancer and have potential for drug sensitivity testing. Previously, we created gastric cancer PDOs from esophagogastroduodenoscopy (EGD) biopsy specimens. As a continuation of our previous work, we sought to optimize methods for creating EGD-derived PDOs for drug sensitivity testing within clinically actionable time constraints.
We enrolled patients with gastric adenocarcinoma undergoing EGD at an NCI cancer center. During diagnostic EGDs, additional research biopsies were collected for creation of gastric cancer PDOs. Briefly, tissues were isolated and washed in medium. After isolation, tissues were placed on a glass slide under the microscope and gentle pressure was applied to release gastric glands. The glands were collected in medium and large tissue fragments were allowed to settle. Using a modification to our earlier techniques, large tissue fragments were isolated, washed, and harvested a second time to further capture glands. These secondary glands were plated separately on a 24-well plate and PDO growth from first and second isolations were observed. On day 5, PDOs were passaged. After the second passage on day 10, PDOs were plated on a 48-well plate and treated with cytotoxic combination chemotherapies, FLOT, MAGIC, and FOLFOX. Cell Titer Glo assay was performed to measure response to chemotherapies. Furthermore, we tested the feasibility of creating EGD-derived PDOs following overnight shipment of specimens.
Consistent with our prior experience, we were able to successfully develop EGD-derived gastric cancer PDOs from patients undergoing diagnostic EGD. Using our modified technique for PDO creation, we established over double the concentration of PDOs from a second isolation. PDOs were passaged and expanded into additional wells on day 5 and day 10 and then treated with current standard of care chemotherapy regimens to predict in vivo drug response. After overnight shipment of EGD biopsy specimens to our lab, we successfully established PDOs at high confluence. Creation of all EGD-derived PDOs and drug sensitivity testing was completed within two weeks of tissue collection.
We have further optimized methodology for EGD-derived PDO creation for immediate drug testing. Creating PDOs in non-optimal conditions from overnight shipment suggests it is feasible to establish these models and conduct drug testing in a timely manner for patients at outside institutions. Furthermore, our modified technique yielded higher PDO concentration, providing sufficient biological material for future implementation of rapid and comprehensive drug testing for neoadjuvant chemotherapy trials.
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