Abstract A14: Engineering pancreatic cancer through molecular and cellular reconstruction

Pancreatic ductal adenocarcinoma (PDAC) is one of the most deadly human malignancies characterized by the accumulation of a series of genetic mutations during disease progression. Despite significant amount of studies using genetically engineered mouse PDAC models, human PDAC still has a dismal prognosis, and there are few models for human PDAC tumorigenesis. To establish a genetic modification system for human pancreatic tissue, we made lentiviral CRISPR to target CDKN2A/p16, p53 and SMAD4. We have confirmed the efficiency of the p16 CRISPR, p53 CRISPR and SMAD4 CRISPR constructs in editing the genome to delete the respective gene loci. We then generated a lentiviral vector to express KRASG12D-mCherry and transduced normal human primary pancreatic exocrine cells with single or multiple viral vector combinations. After one week of culture, two million of the cells were transplanted into the renal sub-capsular space of NOD-SCID mice. The engraftments were harvested 4 months after transplantation. We observed two PanIN lesions in kidneys where KRASG12D mutant cells were transplanted. Human cells were detected using anti-human nuclear antigen antibody (anti-HuNu). Anti-Pan cytokeratin staining proved that the lesions were made up of epithelial cells. The histologic appearance of the lesion from the cells with all four mutations, KRASG12D p16-/- p53-/- SMAD4-/-, differed from that of the two PanIN lesions in that has a single, much larger cyst with mucin rich cytoplasm reminiscent of mucinous cystic neoplasm (MCN). To further define the cellular origin of PDAC initiation, we also developed a flow cytometry-based method to sort viable ductal cells and acinar cells from primary normal human pancreatic cells for genetic modifications. In summary, we have successfully generated PDAC-associated-precancerous lesions from primary normal human pancreatic tissues. The system established in this study could be a valuable platform to address the fundamental mechanisms for the human PDAC tumorigenesis process. Citation Format: Naoki Akanuma, Jun Liu, Pei Wang.{Authors}. Engineering pancreatic cancer through molecular and cellular reconstruction. [abstract]. In: Proceedings of the AACR Special Conference on Pancreatic Cancer: Advances in Science and Clinical Care; 2016 May 12-15; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2016;76(24 Suppl):Abstract nr A14.