Ex vivo culture of intact human patient derived pancreatic tumour tissue

John Kokkinos; George Sharbeen; Koroush S. Haghighi; Rosa Mistica C. Ignacio; Chantal Kopecky; Estrella Gonzales-Aloy; Janet Youkhana; Paul Timpson; Brooke A. Pereira; Shona Ritchie; Elvis Pandzic; Cyrille Boyer; Thomas P. Davis; Lisa M. Butler; David Goldstein; Joshua A. McCarroll; Phoebe A. Phillips

doi:10.1038/s41598-021-81299-0

Ex vivo culture of intact human patient derived pancreatic tumour tissue

John Kokkinos, George Sharbeen, Koroush S. Haghighi, Rosa Mistica C. Ignacio, Chantal Kopecky, Estrella Gonzales-Aloy, Janet Youkhana, Paul Timpson, Brooke A. Pereira, Shona Ritchie, Elvis Pandzic, Cyrille Boyer, Thomas P. Davis, Lisa M. Butler, David Goldstein, Joshua A. McCarroll, Phoebe A. Phillips

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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 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 of human pancreatic tumours. Patient-derived surgically-resected PDAC tissue was cut into 1–2 mm explants and cultured on gelatin sponges for 12 days. Immunohistochemistry revealed that human PDAC explants were viable for 12 days and maintained their original tumour, stromal and extracellular matrix architecture. As proof-of-principle, human PDAC explants were treated with Abraxane and we observed different levels of response between patients. PDAC explants were also transfected with polymeric nanoparticles + Cy5-siRNA and we observed abundant cytoplasmic distribution of Cy5-siRNA throughout the PDAC explants. Overall, our novel model retains the 3D architecture of human PDAC and has advantages over standard organoids: presence of functional multi-cellular stroma and fibrosis, and no tissue manipulation, digestion, or artificial propagation of organoids. This provides unprecedented opportunity to study PDAC biology including tumour-stromal interactions and rapidly assess therapeutic response to drive personalised treatment.

Original language	English
Article number	1944
Number of pages	15
Journal	Scientific Reports
Volume	11
Issue number	1
DOIs	https://doi.org/10.1038/s41598-021-81299-0
Publication status	Published - Dec 2021

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Kokkinos, J., Sharbeen, G., Haghighi, K. S., Ignacio, R. M. C., Kopecky, C., Gonzales-Aloy, E., Youkhana, J., Timpson, P., Pereira, B. A., Ritchie, S., Pandzic, E., Boyer, C., Davis, T. P., Butler, L. M., Goldstein, D., McCarroll, J. A., & Phillips, P. A. (2021). Ex vivo culture of intact human patient derived pancreatic tumour tissue. Scientific Reports, 11(1), Article 1944. https://doi.org/10.1038/s41598-021-81299-0

@article{5d0a06ffd15b4e29b140f9b469cce774,

title = "Ex vivo culture of intact human patient derived pancreatic tumour tissue",

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 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 of human pancreatic tumours. Patient-derived surgically-resected PDAC tissue was cut into 1–2 mm explants and cultured on gelatin sponges for 12 days. Immunohistochemistry revealed that human PDAC explants were viable for 12 days and maintained their original tumour, stromal and extracellular matrix architecture. As proof-of-principle, human PDAC explants were treated with Abraxane and we observed different levels of response between patients. PDAC explants were also transfected with polymeric nanoparticles + Cy5-siRNA and we observed abundant cytoplasmic distribution of Cy5-siRNA throughout the PDAC explants. Overall, our novel model retains the 3D architecture of human PDAC and has advantages over standard organoids: presence of functional multi-cellular stroma and fibrosis, and no tissue manipulation, digestion, or artificial propagation of organoids. This provides unprecedented opportunity to study PDAC biology including tumour-stromal interactions and rapidly assess therapeutic response to drive personalised treatment. ",

author = "John Kokkinos and George Sharbeen and Haghighi, {Koroush S.} and Ignacio, {Rosa Mistica C.} and Chantal Kopecky and Estrella Gonzales-Aloy and Janet Youkhana and Paul Timpson and Pereira, {Brooke A.} and Shona Ritchie and Elvis Pandzic and Cyrille Boyer and Davis, {Thomas P.} and Butler, {Lisa M.} and David Goldstein and McCarroll, {Joshua A.} and Phillips, {Phoebe A.}",

note = "Funding Information: This research was made possible by a Tour de Cure PhD Support Scholarship (Kokkinos, Phillips, Goldstein, RSP-011-18/19) and a Cancer Institute NSW {\textquoteleft}The Professor Rob Sutherland AO Make a Difference Award{\textquoteright} (Goldstein, 2017/AWD002). We also acknowledge the generous philanthropic support of Mr Paul Dainty, Dr Marjorie O{\textquoteright}Neil, Dr Keri Spooner, and the Churches Football Association Sydney (CFA Sydney). The following sources supported author contributions and research: Australian Government Research Training Program Scholarship and UNSW Sydney Scientia PhD Scholarship (Kokkinos), NHMRC project grant (Phillips, McCarroll, Goldstein, APP1144108) and the Avner Pancreatic Cancer Foundation Innovation Grant (Phillips, McCarroll, Goldstein, Davis and Sharbeen, APCF0050618), Cancer-Institute NSW ECF/CDFs (Sharbeen, CDF181166), Olivia Lambert Foundation (McCarroll), Cancer Australia (Phillips, McCarroll and Goldstein, APP1126736), UNSW Medicine Cancer Theme and SPHERE Cancer Clinical Academic Group E/MCR Seed Grant (Kopecky), UNSW International Postgraduate Award (Ritchie), Sydney Catalyst Seed Funding Grant (Pereira), Lens Ains-worth Fellowship in Pancreatic Cancer Research (Timpson), NHMRC Senior Research Fellowship (Timpson, APP1136974), Avner Pancreatic Cancer Foundation Grant (Timpson) and Beat Cancer Project Principal Cancer Research Fellowship (Butler, PRF1117). Publisher Copyright: {\textcopyright} 2021, The Author(s). Copyright: Copyright 2021 Elsevier B.V., All rights reserved.",

year = "2021",

month = dec,

doi = "10.1038/s41598-021-81299-0",

language = "English",

volume = "11",

journal = "Scientific Reports",

issn = "2045-2322",

publisher = "Nature Publishing Group",

number = "1",

}

Kokkinos, J, Sharbeen, G, Haghighi, KS, Ignacio, RMC, Kopecky, C, Gonzales-Aloy, E, Youkhana, J, Timpson, P, Pereira, BA, Ritchie, S, Pandzic, E, Boyer, C, Davis, TP, Butler, LM, Goldstein, D, McCarroll, JA & Phillips, PA 2021, 'Ex vivo culture of intact human patient derived pancreatic tumour tissue', Scientific Reports, vol. 11, no. 1, 1944. https://doi.org/10.1038/s41598-021-81299-0

TY - JOUR

T1 - Ex vivo culture of intact human patient derived pancreatic tumour tissue

AU - Kokkinos, John

AU - Sharbeen, George

AU - Haghighi, Koroush S.

AU - Ignacio, Rosa Mistica C.

AU - Kopecky, Chantal

AU - Gonzales-Aloy, Estrella

AU - Youkhana, Janet

AU - Timpson, Paul

AU - Pereira, Brooke A.

AU - Ritchie, Shona

AU - Pandzic, Elvis

AU - Boyer, Cyrille

AU - Davis, Thomas P.

AU - Butler, Lisa M.

AU - Goldstein, David

AU - McCarroll, Joshua A.

AU - Phillips, Phoebe A.

N1 - Funding Information: This research was made possible by a Tour de Cure PhD Support Scholarship (Kokkinos, Phillips, Goldstein, RSP-011-18/19) and a Cancer Institute NSW ‘The Professor Rob Sutherland AO Make a Difference Award’ (Goldstein, 2017/AWD002). We also acknowledge the generous philanthropic support of Mr Paul Dainty, Dr Marjorie O’Neil, Dr Keri Spooner, and the Churches Football Association Sydney (CFA Sydney). The following sources supported author contributions and research: Australian Government Research Training Program Scholarship and UNSW Sydney Scientia PhD Scholarship (Kokkinos), NHMRC project grant (Phillips, McCarroll, Goldstein, APP1144108) and the Avner Pancreatic Cancer Foundation Innovation Grant (Phillips, McCarroll, Goldstein, Davis and Sharbeen, APCF0050618), Cancer-Institute NSW ECF/CDFs (Sharbeen, CDF181166), Olivia Lambert Foundation (McCarroll), Cancer Australia (Phillips, McCarroll and Goldstein, APP1126736), UNSW Medicine Cancer Theme and SPHERE Cancer Clinical Academic Group E/MCR Seed Grant (Kopecky), UNSW International Postgraduate Award (Ritchie), Sydney Catalyst Seed Funding Grant (Pereira), Lens Ains-worth Fellowship in Pancreatic Cancer Research (Timpson), NHMRC Senior Research Fellowship (Timpson, APP1136974), Avner Pancreatic Cancer Foundation Grant (Timpson) and Beat Cancer Project Principal Cancer Research Fellowship (Butler, PRF1117). Publisher Copyright: © 2021, The Author(s). Copyright: Copyright 2021 Elsevier B.V., All rights reserved.

PY - 2021/12

Y1 - 2021/12

N2 - 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 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 of human pancreatic tumours. Patient-derived surgically-resected PDAC tissue was cut into 1–2 mm explants and cultured on gelatin sponges for 12 days. Immunohistochemistry revealed that human PDAC explants were viable for 12 days and maintained their original tumour, stromal and extracellular matrix architecture. As proof-of-principle, human PDAC explants were treated with Abraxane and we observed different levels of response between patients. PDAC explants were also transfected with polymeric nanoparticles + Cy5-siRNA and we observed abundant cytoplasmic distribution of Cy5-siRNA throughout the PDAC explants. Overall, our novel model retains the 3D architecture of human PDAC and has advantages over standard organoids: presence of functional multi-cellular stroma and fibrosis, and no tissue manipulation, digestion, or artificial propagation of organoids. This provides unprecedented opportunity to study PDAC biology including tumour-stromal interactions and rapidly assess therapeutic response to drive personalised treatment.

AB - 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 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 of human pancreatic tumours. Patient-derived surgically-resected PDAC tissue was cut into 1–2 mm explants and cultured on gelatin sponges for 12 days. Immunohistochemistry revealed that human PDAC explants were viable for 12 days and maintained their original tumour, stromal and extracellular matrix architecture. As proof-of-principle, human PDAC explants were treated with Abraxane and we observed different levels of response between patients. PDAC explants were also transfected with polymeric nanoparticles + Cy5-siRNA and we observed abundant cytoplasmic distribution of Cy5-siRNA throughout the PDAC explants. Overall, our novel model retains the 3D architecture of human PDAC and has advantages over standard organoids: presence of functional multi-cellular stroma and fibrosis, and no tissue manipulation, digestion, or artificial propagation of organoids. This provides unprecedented opportunity to study PDAC biology including tumour-stromal interactions and rapidly assess therapeutic response to drive personalised treatment.

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U2 - 10.1038/s41598-021-81299-0

DO - 10.1038/s41598-021-81299-0

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AN - SCOPUS:85099771330

SN - 2045-2322

VL - 11

JO - Scientific Reports

JF - Scientific Reports

IS - 1

M1 - 1944

ER -

Ex vivo culture of intact human patient derived pancreatic tumour tissue

Abstract

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ARC Centre of Excellence in Convergent Bio-Nano Science and Technology

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Ex vivo culture of intact human patient derived pancreatic tumour tissue

Abstract

UN SDGs

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ARC Centre of Excellence in Convergent Bio-Nano Science and Technology

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