Designer pancreatic cancer generated from human pluripotent stem cell derived ducts

Engineering approaches to build human pancreatic tissue resembling acinar, ductal and endocrine tissue have been hampered by the complexity of the pancreas. Human pluripotent stem cells (PSCs) may provide the appropriate bioengineering platform for developmental and biomedical studies due to their capability to differentiate into every cell type in the human body. However, PSCs typically yield heterogeneous population, while certain disease models require homogenous populations. Here, we have implemented signals controlling embryonic lineage fate bifurcations to efficiently yield the desired cell types through exclusion of alternate fates: Specifically, we applied signaling molecules and growth factors inducing ductal cells, while inhibiting the respective counter lineage with inhibitors. Afterwards, we genome edited human PSCs to specifically design the core machinery of the pancreatic cancer genome. Orthotopic transplantation was additionally incorporated into the experimental work flow. This approach yields virtually pure pancreatic duct-like cells generated from human PSCs resembling key features of adult human pancreatic ducts: A robust test battery including functional Carbanhydrase and CFTR activity and homogenous ductal marker expression underpinned ductal identity. Transcriptional profiles of engineered ducts match the one of human mature ducts. In line orthotopic transplantation reveals human untransformed ducts in the murine host. Pluripotent stem cells armed with oncogenic KRAS were similarly differentiated to the ductal lineage followed by oncogene activation in vitro and in vivo. Here, mutant KRAS alters ductal organoid polarity as early sign of dysplastic growth in vitro, followed by the generation of moderately differentiated human PDAC in vivo. Summarized, this novel and unique differentiation platform generates human untransformed ducts allowing modelling of plasticity, dysplasia and cancer formation in a human and genetically defined background in the pancreas. Thereby, we also show that human ducts are indeed permissive to generate human PDAC. Tailored genome editing strategies mimicking the mutational make-up of human PDAC will open novel opportunities to provide a unique and valuable human PDAC model.