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

Abstract

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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References

  1. 1.

    Siegel RL, Miller KD, Jemal A. Cancer statistics, 2019. CA Cancer J Clin. 2019;69:7–34.

    PubMed  PubMed Central  Google Scholar 

  2. 2.

    Hruban RH, Gaida MM, Thompson E, Hong SM, Noë M, Brosens LA, et al. Why is pancreatic cancer so deadly? The pathologist’s view. J Pathol. 2019;248:131–41.

    Article  Google Scholar 

  3. 3.

    Hong SM, Goggins M, Wolfgang CL, Schulick RD, Edil BH, Cameron JL, et al. Vascular invasion in infiltrating ductal adenocarcinoma of the pancreas can mimic pancreatic intraepithelial neoplasia: a histopathologic study of 209 cases. Am J Surg Pathol. 2012;36:235–41.

    Article  Google Scholar 

  4. 4.

    Yamada M, Sugiura T, Okamura Y, Ito T, Yamamoto Y, Ashida R, et al. Microscopic venous invasion in pancreatic cancer. Ann Surg Oncol. 2018;25:1043–51.

    Article  Google Scholar 

  5. 5.

    Suenaga M, Fujii T, Kanda M, Takami H, Okumura N, Inokawa Y, et al. Pattern of first recurrent lesions in pancreatic cancer: hepatic relapse is associated with dismal prognosis and portal vein invasion. Hepatogastroenterology. 2014;61:1756–61.

    PubMed  Google Scholar 

  6. 6.

    Hamada Y, Nakayama Y. Aggressive venous invasion in the area of carcinoma correlates with liver metastasis as an index of metastasis for invasive ductal carcinoma of the pancreas. Pancreatology. 2017;17:951–5.

    Article  Google Scholar 

  7. 7.

    Hong SM, Noë M, Hruban CA, Thompson ED, Wood LD, Hruban RHA. “Clearer” view of pancreatic pathology: a review of tissue clearing and advanced microscopy techniques. Adv Anat Pathol. 2019;26:31–9.

    Article  CAS  Google Scholar 

  8. 8.

    Noë M, Rezaee N, Asrani K, Skaro M, Groot VP, Wu PH, et al. Immunolabeling of cleared human pancreata provides insights into three-dimensional pancreatic anatomy and pathology. Am J Pathol. 2018;188:1530–5.

    Article  Google Scholar 

  9. 9.

    Sharma S, Green KB. The pancreatic duct and its arteriovenous relationship: an underutilized aid in the diagnosis and distinction of pancreatic adenocarcinoma from pancreatic intraepithelial neoplasia. A study of 126 pancreatectomy specimens. Am J Surg Pathol. 2004;28:613–20.

    Article  Google Scholar 

  10. 10.

    Jeanes A, Gottardi CJ, Yap AS. Cadherins and cancer: how does cadherin dysfunction promote tumor progression? Oncogene. 2008;27:6920–9.

    Article  CAS  Google Scholar 

  11. 11.

    Onder TT, Gupta PB, Mani SA, Yang J, Lander ES, Weinberg RA. Loss of E-cadherin promotes metastasis via multiple downstream transcriptional pathways. Cancer Res. 2008;68:3645–54.

    Article  CAS  Google Scholar 

  12. 12.

    Cao D, Maitra A, Saavedra JA, Klimstra DS, Adsay NV, Hruban RH. Expression of novel markers of pancreatic ductal adenocarcinoma in pancreatic nonductal neoplasms: additional evidence of different genetic pathways. Mod Pathol. 2005;18:752–61.

    Article  CAS  Google Scholar 

  13. 13.

    Winter JM, Ting AH, Vilardell F, Gallmeier E, Baylin SB, Hruban RH, et al. Absence of E-cadherin expression distinguishes noncohesive from cohesive pancreatic cancer. Clin Cancer Res. 2008;14:412–8.

    Article  CAS  Google Scholar 

  14. 14.

    Zeisberg M, Neilson EG. Biomarkers for epithelial-mesenchymal transitions. J Clin Investig. 2009;119:1429–37.

    Article  CAS  Google Scholar 

  15. 15.

    Nguyen DT, Lee E, Alimperti S, Norgard RJ, Wong A, Lee JJ, et al. A biomimetic pancreatic cancer on-chip reveals endothelial ablation via ALK7 signaling. Sci Adv. 2019;5:eaav6789.

    Article  Google Scholar 

  16. 16.

    Gresta LT, Rodrigues-Junior IA, de Castro LP, Cassali GD, Cabral MM. Assessment of vascular invasion in gastric cancer: a comparative study. World J Gastroenterol. 2013;19:3761–9.

    Article  Google Scholar 

  17. 17.

    Inada K, Shimokawa K, Ikeda T, Ozeki Y. The clinical significance of venous invasion in cancer of the stomach. Jpn J Surg. 1990;20:545–52.

    Article  CAS  Google Scholar 

  18. 18.

    Hwang C, Lee S, Kim A, Kim YG, Ahn SJ, Park DY. Venous invasion in colorectal cancer: impact of morphologic findings on detection rate. Cancer Res Treat. 2016;48:1222–8.

    Article  Google Scholar 

  19. 19.

    Betge J, Pollheimer MJ, Lindtner RA, Kornprat P, Schlemmer A, Rehak P, et al. Intramural and extramural vascular invasion in colorectal cancer: prognostic significance and quality of pathology reporting. Cancer. 2012;118:628–38.

    Article  Google Scholar 

  20. 20.

    Yu J, Blackford AL, Dal Molin M, Wolfgang CL, Goggins M. Time to progression of pancreatic ductal adenocarcinoma from low-to-high tumour stages. Gut. 2015;64:1783–9.

    Article  Google Scholar 

  21. 21.

    Nieto MA, Huang RY, Jackson RA, Thiery JP. EMT: 2016. Cell. 2016;166:21–45.

    Article  CAS  Google Scholar 

  22. 22.

    Micalizzi DS, Haber DA, Maheswaran S. Cancer metastasis through the prism of epithelial-to-mesenchymal transition in circulating tumor cells. Mol Oncol. 2017;11:770–80.

    Article  Google Scholar 

  23. 23.

    Chen T, You Y, Jiang H, Wang ZZ. Epithelial-mesenchymal transition (EMT): a biological process in the development, stem cell differentiation, and tumorigenesis. J Cell Physiol. 2017;232:3261–72.

    Article  CAS  Google Scholar 

  24. 24.

    Serrano-Gomez SJ, Maziveyi M, Alahari SK. Regulation of epithelial-mesenchymal transition through epigenetic and post-translational modifications. Mol Cancer. 2016;15:18.

    Article  Google Scholar 

  25. 25.

    Brabletz T, Jung A, Reu S, Porzner M, Hlubek F, Kunz-Schughart LA, et al. Variable beta-catenin expression in colorectal cancers indicates tumor progression driven by the tumor environment. Proc Natl Acad Sci USA. 2001;98:10356–61.

    Article  CAS  Google Scholar 

  26. 26.

    Ligorio M, Sil S, Malagon-Lopez J, Nieman LT, Misale S, Di Pilato M, et al. Stromal microenvironment shapes the intratumoral architecture of pancreatic. Cancer Cell. 2019;178:160–75 e27.

    CAS  Google Scholar 

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Acknowledgements

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 33, 639–647 (2020). https://doi.org/10.1038/s41379-019-0409-3

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