Generation and characterization of a cell line from an intraductal tubulopapillary neoplasm of the pancreas

Abstract

Intraductal tubulopapillary neoplasm (ITPN) is a distinct precancerous lesion in the pancreas with unique clinical and molecular features. Although in vitro studies in two-dimensional culture have led to numerous important insights in pancreatic cancer, such models are currently lacking for precancerous lesions. In this study, we report the generation and characterization of a cell line from a human pancreatic ITPN. Neoplastic cells were initially cultured in a three-dimensional organoid system, followed by transfer to two-dimensional culture. RNA sequencing revealed a gene expression profile consistent with pancreatic ductal origin, and whole genome sequencing identified many somatic mutations (including in genes involved in DNA repair and WNT signaling) and structural rearrangements. In vitro characterization of the tumorigenic potential demonstrated a phenotype between that of normal pancreatic ductal cells and cancer cell lines. This cell line represents a valuable resource for interrogation of unique ITPN biology, as well as precancerous pancreatic lesions more generally.

Molecular characterization of organoids derived from pancreatic intraductal papillary mucinous neoplasms

“…Whole genome sequencing was performed as previously described [25]. DNA from eight IPMN organoids (in passages 1–4) and matched primary non‐neoplastic duodenal tissue were whole genome‐sequenced by the Next Generation Sequencing Core of the Johns Hopkins Sidney Kimmel Comprehensive Cancer Center as previously described [25].…”

“…Whole genome sequencing was performed as previously described [25]. DNA from eight IPMN organoids (in passages 1–4) and matched primary non‐neoplastic duodenal tissue were whole genome‐sequenced by the Next Generation Sequencing Core of the Johns Hopkins Sidney Kimmel Comprehensive Cancer Center as previously described [25]. In brief, sequence libraries were generated using the TruSeq DNA Nano Library Prep Kit (Illumina, San Diego, CA, USA; catalog No 20015964) and sequenced on a HiSeq 2500 (Illumina) to generate 2 × 150 bp reads.…”

“…Whole exome sequencing was performed as previously described [25]. Briefly, paired IPMN (tumor) and duodenum (normal) DNA from H55, H68, and H79 were extracted from FFPE tissue and whole exome‐sequenced by the Next Generation Sequencing Core of the Johns Hopkins Sidney Kimmel Comprehensive Cancer Center.…”

“…Sidney Kimmel Comprehensive Cancer Center as previously described [25]. In brief, sequence libraries were generated using the…”

“…RNA-seq was performed as previously described [25]. IHC results were evaluated using an H-score, and a semi-quantitative, non-blinded approach.…”