Bringing the pancreas patient back to the bench: Ex vivo culture of intact human patient derived pancreatic tumour tissue

Kokkinos, J.; Sharbeen, G.; Haghighi, K. S.; Ignacio, R. M. C.; Kopecky, C.; Gonzales-Aloy, E.; Youkhana, J.; Pandzic, E.; Boyer, C.; Davis, T. P.; Butler, L. M.; Goldstein, D.; McCarroll, J. A.; Phillips, P. A.

doi:10.1101/2020.07.30.223925

bioRxiv · 10.1101/2020.07.30.223925

Bringing the pancreas patient back to the bench: Ex vivo culture of intact human patient derived pancreatic tumour tissue

Kokkinos, J.·Sharbeen, G.·Haghighi, K. S.·Ignacio, R. M. C.·Kopecky, C.·Gonzales-Aloy, E.·Youkhana, J.·Pandzic, E.·Boyer, C.·Davis, T. P.·Butler, L. M.·Goldstein, D.·McCarroll, J. A.·Phillips, P. A.

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Abstract

The poor prognosis of pancreatic ductal adenocarcinoma (PDAC) is attributed to the highly fibrotic stroma and complex multi-cellular microenvironment that is difficult to fully recapitulate in pre-clinical human models. To fast-track translation of therapies and to inform personalised medicine, we aimed to develop a whole-tissue ex vivo explant model that maintains viability, 3D multicellular architecture, and microenvironmental cues present in human pancreatic tumours. Patient-derived surgically-resected PDAC tissue was cut into 2 mm explants, cultured on gelatin sponges, and grown for 12 days. Immunohistochemistry revealed that human PDAC tissue explants were viable for 12 days and maintained their original tumour, stromal and extracellular matrix architecture. As proof-of-principle, human PDAC tissue explants responded to Abraxane(R) treatment with a 3.7-fold increase in cell-death (p=0.0007). PDAC explants were also transfected with polymeric nanoparticles+Cy5-siRNA and we observed abundant cytoplasmic distribution of nanoparticle+Cy5-siRNA throughout the PDAC explant tissue. Our novel model retains the 3D architecture of human pancreatic tumours and has several advantages over standard organoids: presence of functional multi-cellular stroma and fibrosis and no tissue manipulation, digestion, or artificial propagation of organoids. This provides an unprecedented opportunity to study PDAC biology, tumour-stromal interactions and rapidly assess therapeutic response that could drive personalised treatment for PDAC.

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Kokkinos, J., Sharbeen, G., Haghighi, K. S., Ignacio, R. M. C., Kopecky, C., Gonzales-Aloy, E., Youkhana, J., Pandzic, E., Boyer, C., Davis, T. P., Butler, L. M., Goldstein, D., McCarroll, J. A., Phillips, P. A.. 2020-07-30. Bringing the pancreas patient back to the bench: Ex vivo culture of intact human patient derived pancreatic tumour tissue. https://doi.org/10.1101/2020.07.30.223925

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