Feasibility of Organoid-Based Modeling for Human Intraductal Papillary Mucinous Neoplasms of the Pancreas
*Nicholas V Peters, *Prashanth Gokare, *Nesrin Hasan, *Paulomi B Aldo, *Anup R Sharma, Ronald R Salem, Nita Ahuja, John W Kunstman
Yale University, New Haven, CT
Objective: Pancreatic intraductal papillary mucinous neoplasms (IPMN) are premalignant cystic lesions lacking an in vitro model for study, thus limiting the understanding of their progression to adenocarcinoma. Organoid-based systems have emerged as a genetically-tractable approach for tumor study. We sought to assess the feasibility of creating an organoid-based model of human IPMN.
Design: Prospective proof-of-concept
Setting: Translational study, university center
Patients: Patients undergoing surgical resection for IPMN (any subtype)
Interventions: Pancreatectomy specimens were enzymatically digested to oligo-cell clusters, embedded in solubilized basement membrane (Matrigel), and grown to confluence in permissive media promoting enrichment of the ductal epithelial fraction. Microscopy, immunohistochemistry (IHC), and genotypic analysis of known highly-penetrant IPMN mutations were then performed.
Main Outcome Measures: Successful creation of spherical organoids capable of sustained growth, defined as 3 independent culture passages.
Results: 13 unique cyst wall or pancreatic ductal tissue sections were freshly harvested from pancreatectomy specimens derived from 6 patients (range, 1-6 sections/patient) and subsequently confirmed as IPMN via matched histology. Spheroid formation and sustained growth were attained in 5 organoid cultures (38.5%) as demonstrated by phase-contrast microscopy (Figure). Genomic DNA was successfully isolated from 4 organoid cultures and targeted sequencing revealed KRAS G12x/Q61x or GNAS R201x mutations in 3 samples (75%). To date, IHC analysis of 1 culture demonstrated recapitulation of pathognomonic IPMN-defining protein expression consistent with the clinical tissue of origin (CDX2, MUC2, MUC5AC).
Conclusions: Creation of sustained human-derived IPMN organoid culture models is feasible. Future improvements in organoid generation efficiency will enhance the applicability of this approach. 
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