Three-dimensional visualization of cleared human pancreas cancer reveals that sustained epithelial-to-mesenchymal transition is not required for venous invasion

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Venous invasion is three times more common in pancreatic cancer than it is in other major cancers of the gastrointestinal tract, and venous invasion may explain why pancreatic cancer is so deadly. To characterize the patterns of venous invasion in pancreatic cancer, 52 thick slabs (up to 5 mm) of tissue were harvested from 52 surgically resected human ductal adenocarcinomas, cleared with a modified iDISCO method, and labeled with fluorescent-conjugated antibodies to cytokeratin 19, desmin, CD31, p53 and/or e-cadherin. Labeled three-dimensional (3D) pancreas cancer tissues were visualized with confocal laser scanning or light sheet microscopy. Multiple foci of venous and even arterial invasion were visualized. Venous invasion was detected more often in 3D (88%, 30/34 cases) than in conventional 2D slide evaluation (75%, 25/34 cases, P < 0.001). 3D visualization revealed pancreatic cancer cells crossing the walls of veins at multiple points, often at points where preexisting capillary structures bridge the blood vessels. The neoplastic cells often retained a ductal morphology (cohesive cells forming tubes) as they progressed from a stromal to intravenous location. Although immunolabeling with antibodies to e-cadherin revealed focal loss of expression at the leading edges of the cancers, the neoplastic cells within veins expressed e-cadherin and formed well-oriented glands. We conclude that venous invasion is almost universal in pancreatic cancer, suggesting that even surgically resectable PDAC has access to the venous spaces and thus the ability to disseminate widely. Furthermore, we observe that sustained epithelial–mesenchymal transition is not required for venous invasion in pancreatic cancer.

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This work was presented in part at the 2019 annual meeting of United States and Canadian Academy of Pathology, National Harbor, MD. LW is supported by the Sol Goldman Pancreatic Cancer Research Center, Susan Wojcicki and Dennis Troper, the Michael Rolfe Foundation for Pancreatic Cancer Research, Buffone Family Gastrointestinal Cancer Research Fund, Kaya Tuncer Career Development Award in Gastrointestinal Cancer Prevention, AGA -Bernard Lee Schwartz Foundation Research Scholar Award in Pancreatic Cancer, Sidney Kimmel Foundation for Cancer Research Kimmel Scholar Award, AACR-Incyte Corporation Career Development Award for Pancreatic Cancer Research, Joseph C Monastra Foundation, The Gerald O Mann Charitable Foundation (Harriet and Allan Wulfstat, Trustees). MN is supported by The Nijbakker-Morra Foundation, The Lisa Waller Hayes Foundation. SMH is supported by the National Research Foundation of Korea (NRF) funded by the Ministry of Science, ICT, and Future Planning (NRF-2019R1H1A2102016). MMG is supported by the German Research Foundation (Ga 1818/2-1). The authors thank Drs Michael Goggins and Anne Macgregor-Das for providing p53-positive pancreatic ductal adenocarcinomas, Drs Robert A. Anders, Alan K. Meeker, Christopher M. Heaphy, and Scot C. Guo for helpful discussions for this project, and Ms Yuan Kai and Ms Barbara Smith for their technical support.

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Correspondence to Ralph H. Hruban or Laura D. Wood.

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Hong, S., Jung, D., Kiemen, A. et al. Three-dimensional visualization of cleared human pancreas cancer reveals that sustained epithelial-to-mesenchymal transition is not required for venous invasion. Mod Pathol (2019) doi:10.1038/s41379-019-0409-3

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