Abstract 6511: Optimizing an in vitro toolbox to interrogate pancreatic tumorigenesis from a patient-derived intraductal papillary mucinous neoplasm sample

Abstract

Abstract Pancreatic Ductal Adenocarcinoma (PDAC) is a lethal disease with a five-year survival rate of 11%. PDAC develops from pancreatic precursor lesions, including intraductal papillary mucinous neoplasm (IPMN); studying these lesions is important to understand the multi-step process of pancreatic tumorigenesis. Development of model systems and associated tools is a critical step that is 1) scientifically necessary to understand IPMN biology and 2) clinically important for improving patient outcome. To date, only one patient-derived IPMN cell line that grows in a monolayer has been reported, illustrating the challenges of growing precancerous cells in this format. While protocols have previously been established to generate patient-derived IPMN models, the majority of these cell-based models utilize 3D organoids. To date, no study has successfully converted patient-derived IPMN organoids into 2D culture. Converting these cellular models to propagate and divide in a monolayer allows us to expand our in vitro toolbox to perform experimental methods that are less efficient or not practical in 3D culture. In this study, we optimized the conversion of a high-grade IPMN patient-derived organoid model into a monolayer culture. We considered conditions that could recapitulate an extracellular matrix as well as growth factors and/or inhibitors that could promote IPMN cell growth. In total, we tested 15 different conditions and were able to successfully culture the human high-grade IPMN cells in a monolayer culture for up to 5 passages (~1 month). Additionally, while human IPMN cellular models exists, genetic modification of these in vitro models has not been reported. Such approaches allow direct interrogation of the role of specific genes in premalignant pancreatic tumorigenesis. In this study, we report successful transduction of IPMN cells via lentivirus. By first converting the organoid into a monolayer culture, transducing them in 2D, and then converting them back into 3D culture as spheroids, we were able to successfully express EGFP into our IPMN cellular model. In summary, we optimized culture conditions to temporarily grow a patient derived 3D IPMN model in a 2D monolayer form, genetically modified this IPMN cell line model, and successfully converted it back into a 3D spheroid. These protocols have potential to be replicated in other IPMN models and can provide further insight into overall IPMN biology and pancreatic tumorigenesis. Citation Format: Raymond M. Paranal, Julie Schlanz, Maria A. Trujillo, Nicholas J. Roberts, Laura D. Wood. Optimizing an in vitro toolbox to interrogate pancreatic tumorigenesis from a patient-derived intraductal papillary mucinous neoplasm sample [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 6511.