Establishing a Living Biobank of Patient-Derived Organoids of Intraductal Papillary Mucinous Neoplasms of the Pancreas

Abstract

Pancreatic cancer (PaCa) is the third leading cause of cancer-related deaths in the United States. There is an unmet need to develop strategies to detect PaCa at an early, operable stage and prevent its progression. Intraductal papillary mucinous neoplasms (IPMNs) are cystic PaCa precursors that comprise nearly 50% of pancreatic cysts detected incidentally via cross-sectional imaging. Since IPMNs can progress from low- and moderate-grade dysplasia to high-grade dysplasia and invasion, the study of these lesions offers a prime opportunity to develop early detection and prevention strategies. Organoids are an ideal preclinical platform to study IPMNs, and the objective of the current investigation was to establish a living biobank of patient-derived organoids (PDO) from IPMNs. IPMN tumors and adjacent normal pancreatic tissues were successfully harvested from 15 patients with IPMNs undergoing pancreatic surgical resection at Moffitt Cancer Center & Research Institute (Tampa, FL) between May of 2017 and March of 2019. Organoid cultures were also generated from cryopreserved tissues. Organoid count and size were determined over time by both Image-Pro Premier 3D Version 9.1 digital platform and Matlab application of a Circular Hough Transform algorithm, and histologic and genomic characterization of a subset of the organoids was performed using immunohistochemistry and targeted sequencing, respectively. The success rates for organoid generation from IPMN tumor and adjacent normal pancreatic tissues were 81% and 87%, respectively. IPMN organoids derived from different epithelial subtypes showed different morphologies in vitro, and organoids recapitulated histologic and genomic characteristics of the parental IPMN tumor. In summary, this pre-clinical model has the potential to provide new opportunities to unveil mechanisms of IPMN progression to invasion and to shed insight into novel biomarkers for early detection and targets for chemoprevention.

In vitro models for head and neck cancer: Current status and future perspective

“…The tumor tissues were first digested with a proteolytic enzyme for cell retrieval. The recovered cancer cells were then seeded in Matrigel ® and stored as PDOs in a living biobank (119). The gene analysis data of key markers KRAS, PTEN, PIK3CA, GNAS, RNF43, and BRAF showed a similar expression between PDO and the patient's tumor tissue biopsy, which confirmed the preservation of patient samples' genome in living biobanks.…”

“…Fortunately, PDOs can be replicated and cryopreserved in specialized facilities, known as living biobanks, without losing cell-type specificity (87,118). For example, intraductal papillary mucinous neoplasms were collected from patients with pancreatic cancer (119). The tumor tissues were first digested with a proteolytic enzyme for cell retrieval.…”

The Florida Pancreas Collaborative Next-Generation Biobank: State-wide Infrastructure to Reduce Disparities and Improve Survival for a Racially and Ethnically Diverse Cohort of Patients with Pancreatic Cancer

“…We will also be performing genotyping for ancestry informative markers using DNA from blood collected via this study to validate self-reported ancestry. Finally, based on the promise of pre-clinical models in translational efforts [79][80][81][82][83] , we also plan to leverage the pancreatic tissue collected and preserved through this effort for applications such as generation of patient-derived organoids 105 . Thus, the biospecimens collected by the FPC will become an enduring resource for the biomedical and disparities research community.…”