Z Gastroenterol 2019; 57(01): e57
DOI: 10.1055/s-0038-1677198
4. Tumors
Georg Thieme Verlag KG Stuttgart · New York

Verification of 3D liver organoids from cholangiocarcinoma tissue for chemotherapeutic drug screening

S Brückner
1   Clinics of Visceral, Transplantation, Thoracic and Vascular Surgery, University of Leipzig, Leipzig, Germany
,
M Koch
2   Physical Biology, Buchmann Institute for Molecular Life Sciences (BMLS), Goethe-Universität Frankfurt am Main, Germany
,
R Lieshout
3   Department of Surgery, Erasmus MC-University Medical Center, Rotterdam, The Netherlands
,
M Verstegen
3   Department of Surgery, Erasmus MC-University Medical Center, Rotterdam, The Netherlands
,
L van der Laan
3   Department of Surgery, Erasmus MC-University Medical Center, Rotterdam, The Netherlands
,
F Pampaloni
2   Physical Biology, Buchmann Institute for Molecular Life Sciences (BMLS), Goethe-Universität Frankfurt am Main, Germany
,
B Christ
1   Clinics of Visceral, Transplantation, Thoracic and Vascular Surgery, University of Leipzig, Leipzig, Germany
› Author Affiliations
Further Information

Publication History

Publication Date:
04 January 2019 (online)

Also available at
 

Background:

Liver cancer is the fourth most frequent cancer-related cause of death worldwide. Thus, sufficiently effective chemotherapeutic approaches are vitally needed to improve patients' survival. The heterogeneity of liver tumours requires a patient-adjusted chemotherapy regimen.

Aim:

It was the aim to investigate, whether individual patient-derived liver tumor cell organoids invitro were feasible to screen for tumor-specific chemotherapeutics.

Methods:

Functional liver cell markers (PKL, PKM, CYP3A4, CYP1A2, GST, MDR, E-cad, ZO-1) were analyzed by immunohistochemistry and verified by qRT-PCR in tumor and tumor-adjacent tissue. Organoids from individual patient cholangiocarcinoma- were generated and analyzed in culture after Sorafenib (2µM and 4µM) treatment using semi-automatic image-size measurement and immunofluorescence staining (EpCAM, SOX9, MDR, E-cadherin and ZO-1). The proliferation rate of organoids was quantified by Ki67 expression. Organoids derived from healthy liver tissue served as a control.

Results:

Liver tumor and adjacent tissue of 5 patients was analyzed confirming the hepatocellular or cholangiocellular origin of the tumors. However, gene expression patterns featured highly individual differences. This pointed out the importance to unravel tumor-specific chemosensitivity in order to optimize patient-tailored chemotherapeutic treatment efficiency.

All raised organoids stained highly positive for EpCAM. In addition, cells within the organoids were polarized. E-cadherin was expressed on the outside of the organoids, ZO-1 and MDR on the luminal side. The expression pattern of E-cadherin, ZO-1 and MDR was not affected by Sorafenib (2µM and 4µM) treatment. The semi-automatic image-size measurement revealed a reduction of tumor organoid size after 48h of Sorafenib (4µM) treatment. In contrast, Sorafinib did not affect growth of organoids from healthy donors. Interestingly, no differences in the proliferation rate were observed in control organoids or organoids treated with Sorafenib (2µM and 4µM). This indicated that Sorafenib ameliorated organoid size increase, albeit not due to inhibition of cell proliferation in the organoids.

Conclusion:

Patient-specific cholangiocarcinoma-derived organoids may be used for in vitro chemotherapeutic drug screening, which is highly appreciable to tailor patient-specific, optimized chemotherapeutic treatment.