CDKN2A-mutated pancreatic ductal organoids from induced pluripotent stem cells to model a cancer predisposition syndrome

Simple Summary The mutational landscape of pancreatic ductal adenocarcinoma has been delineated in large clinical studies. Such retrospective characterization, however, is limited to late stages of the disease, and exploring the complexity of carcinogenic routes is in its infancy. Here, we provide an in vitro and in vivo model system to directly investigate human PDAC development after the induction of specific oncogenic insults in a cancer-prone ancestry. We reprogrammed plucked human hair-derived keratinocytes from two siblings harboring a pathogenic CDKN2A variant putatively predisposing for PDAC to induced pluripotent stem cells (iPSCs) and additionally introduced an inducible KRASG12D piggyBac expression cassette. Upon KRASG12D induction in differentiated pancreatic duct-like organoids, structural and molecular changes were observed. After orthotopic transplantation either a high-grade precancer lesion or PDAC-like tumor developed. Hereby, we provide a hereditary human pancreatic cancer model which enables further dissection of tumor initiation and early development starting from patient-specific CDKN2A-mutated pluripotent stem cells. Abstract Patient-derived induced pluripotent stem cells (iPSCs) provide a unique platform to study hereditary disorders and predisposition syndromes by resembling germline mutations of affected individuals and by their potential to differentiate into nearly every cell type of the human body. We employed plucked human hair from two siblings with a family history of cancer carrying a pathogenic CDKN2A variant, P16-p.G101W/P14-p.R115L, to generate patient-specific iPSCs in a cancer-prone ancestry for downstream analytics. The differentiation capacity to pancreatic progenitors and to pancreatic duct-like organoids (PDLOs) according to a recently developed protocol remained unaffected. Upon inducible expression of KRASG12D using a piggyBac transposon system in CDKN2A-mutated PDLOs, we revealed structural and molecular changes in vitro, including disturbed polarity and ...