Label-Free Imaging Analysis of Patient-Derived Cholangiocarcinoma Organoids after Sorafenib Treatment

Michael Koch; Sandra Nickel; Ruby Lieshout; Susanna M Lissek; Martina Leskova; Luc J W van der Laan; Monique M A Verstegen; Bruno Christ; Francesco Pampaloni

doi:10.3390/cells11223613

Label-Free Imaging Analysis of Patient-Derived Cholangiocarcinoma Organoids after Sorafenib Treatment

Michael Koch, Sandra Nickel, Ruby Lieshout, Susanna M Lissek, Martina Leskova, Luc J W van der Laan, Monique M A Verstegen, Bruno Christ, Francesco Pampaloni

Surgery

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Abstract

Monitoring tumor growth dynamics is crucial for understanding cancer. To establish an in vitro method for the continuous assessment of patient-specific tumor growth, tumor organoids were generated from patients with intrahepatic CCA (iCCA). Organoid growth was monitored for 48 h by label-free live brightfield imaging. Growth kinetics were calculated and validated by MTS assay as well as immunohistochemistry of Ki67 to determine proliferation rates. We exposed iCCA organoids (iCCAOs) and non-tumor intrahepatic cholangiocyte organoids (ICOs) to sub-therapeutic concentrations of sorafenib. Monitoring the expansion rate of iCCAOs and ICOs revealed that iCCAO growth was inhibited by sorafenib in a time- and dose-dependent fashion, while ICOs were unaffected. Quantification of the proliferation marker Ki67 confirmed inhibition of iCCAO growth by roughly 50% after 48 h of treatment with 4 µM sorafenib. We established a robust analysis pipeline combining brightfield microscopy and a straightforward image processing approach for the label-free growth monitoring of patient-derived iCCAOs. Combined with bioanalytical validation, this approach is suitable for a fast and efficient high-throughput drug screening in tumor organoids to develop patient-specific systemic treatment options.

Original language	English
Article number	3613
Journal	Cells
Volume	11
Issue number	22
DOIs	https://doi.org/10.3390/cells11223613
Publication status	Published - 15 Nov 2022

Bibliographical note

Funding Information:
Results presented were generated in the frame of the consortium “The Onconoid Hub”, which was supported through funding by the German Federal Ministry of Education and Research (BMBF; file no. 031L0022A (BC), 031L0022B (FP)) and The Netherlands Organization for Health Research and Development (ZonMw), project #114027003 (L.v.d.L. and M.M.A.V.) in the InnoSysTox joint call (file no. 114027003). L.v.d.L. and M.M.A.V. are further supported by the Medical Delta program grant (Regenerative Medicine 4D) and the Erasmus MC Human Disease Model Award (HDMA-380801). The APC was funded by the Open Access Publication fund of the Goethe University Library.

Publisher Copyright:
© 2022 by the authors.

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Label-Free Imaging Analysis of Patient-Derived Cholangiocarcinoma Organoids after Sorafenib TreatmentFinal published version, 1.67 MBLicence: CC BY

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title = "Label-Free Imaging Analysis of Patient-Derived Cholangiocarcinoma Organoids after Sorafenib Treatment",

abstract = "Monitoring tumor growth dynamics is crucial for understanding cancer. To establish an in vitro method for the continuous assessment of patient-specific tumor growth, tumor organoids were generated from patients with intrahepatic CCA (iCCA). Organoid growth was monitored for 48 h by label-free live brightfield imaging. Growth kinetics were calculated and validated by MTS assay as well as immunohistochemistry of Ki67 to determine proliferation rates. We exposed iCCA organoids (iCCAOs) and non-tumor intrahepatic cholangiocyte organoids (ICOs) to sub-therapeutic concentrations of sorafenib. Monitoring the expansion rate of iCCAOs and ICOs revealed that iCCAO growth was inhibited by sorafenib in a time- and dose-dependent fashion, while ICOs were unaffected. Quantification of the proliferation marker Ki67 confirmed inhibition of iCCAO growth by roughly 50% after 48 h of treatment with 4 µM sorafenib. We established a robust analysis pipeline combining brightfield microscopy and a straightforward image processing approach for the label-free growth monitoring of patient-derived iCCAOs. Combined with bioanalytical validation, this approach is suitable for a fast and efficient high-throughput drug screening in tumor organoids to develop patient-specific systemic treatment options.",

author = "Michael Koch and Sandra Nickel and Ruby Lieshout and Lissek, {Susanna M} and Martina Leskova and {van der Laan}, {Luc J W} and Verstegen, {Monique M A} and Bruno Christ and Francesco Pampaloni",

note = "Funding Information: Results presented were generated in the frame of the consortium “The Onconoid Hub”, which was supported through funding by the German Federal Ministry of Education and Research (BMBF; file no. 031L0022A (BC), 031L0022B (FP)) and The Netherlands Organization for Health Research and Development (ZonMw), project #114027003 (L.v.d.L. and M.M.A.V.) in the InnoSysTox joint call (file no. 114027003). L.v.d.L. and M.M.A.V. are further supported by the Medical Delta program grant (Regenerative Medicine 4D) and the Erasmus MC Human Disease Model Award (HDMA-380801). The APC was funded by the Open Access Publication fund of the Goethe University Library. Publisher Copyright: {\textcopyright} 2022 by the authors.",

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AU - van der Laan, Luc J W

AU - Verstegen, Monique M A

AU - Christ, Bruno

AU - Pampaloni, Francesco

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N2 - Monitoring tumor growth dynamics is crucial for understanding cancer. To establish an in vitro method for the continuous assessment of patient-specific tumor growth, tumor organoids were generated from patients with intrahepatic CCA (iCCA). Organoid growth was monitored for 48 h by label-free live brightfield imaging. Growth kinetics were calculated and validated by MTS assay as well as immunohistochemistry of Ki67 to determine proliferation rates. We exposed iCCA organoids (iCCAOs) and non-tumor intrahepatic cholangiocyte organoids (ICOs) to sub-therapeutic concentrations of sorafenib. Monitoring the expansion rate of iCCAOs and ICOs revealed that iCCAO growth was inhibited by sorafenib in a time- and dose-dependent fashion, while ICOs were unaffected. Quantification of the proliferation marker Ki67 confirmed inhibition of iCCAO growth by roughly 50% after 48 h of treatment with 4 µM sorafenib. We established a robust analysis pipeline combining brightfield microscopy and a straightforward image processing approach for the label-free growth monitoring of patient-derived iCCAOs. Combined with bioanalytical validation, this approach is suitable for a fast and efficient high-throughput drug screening in tumor organoids to develop patient-specific systemic treatment options.

AB - Monitoring tumor growth dynamics is crucial for understanding cancer. To establish an in vitro method for the continuous assessment of patient-specific tumor growth, tumor organoids were generated from patients with intrahepatic CCA (iCCA). Organoid growth was monitored for 48 h by label-free live brightfield imaging. Growth kinetics were calculated and validated by MTS assay as well as immunohistochemistry of Ki67 to determine proliferation rates. We exposed iCCA organoids (iCCAOs) and non-tumor intrahepatic cholangiocyte organoids (ICOs) to sub-therapeutic concentrations of sorafenib. Monitoring the expansion rate of iCCAOs and ICOs revealed that iCCAO growth was inhibited by sorafenib in a time- and dose-dependent fashion, while ICOs were unaffected. Quantification of the proliferation marker Ki67 confirmed inhibition of iCCAO growth by roughly 50% after 48 h of treatment with 4 µM sorafenib. We established a robust analysis pipeline combining brightfield microscopy and a straightforward image processing approach for the label-free growth monitoring of patient-derived iCCAOs. Combined with bioanalytical validation, this approach is suitable for a fast and efficient high-throughput drug screening in tumor organoids to develop patient-specific systemic treatment options.

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JO - Cells

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ER -

Label-Free Imaging Analysis of Patient-Derived Cholangiocarcinoma Organoids after Sorafenib Treatment

Abstract

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