Abstract 4263: Establishment and initial characterization of human hepatocellular carcinoma organoid culture
Authors: Schölch Sebastian , Nanduri Lahiri K. , Kühn Daniel , Weitz Jürgen , Koch Moritz , Rahbari Nuh N. 
Published: July 2016

Background:

Hepatocellular carcinoma (HCC) is the 3rd most leading cause of cancer death in the world. Despite extensive research, HCC has a strikingly bad prognosis due to the limited therapeutic options: Other than surgical resection, there is no curative treatment. No cytotoxic agent has shown efficacy so far, and the only currently approved systemic agent, sorafenib, is only used in a palliative context.

This dilemma is due to the therapy-resistant biology as well as its variable pathogenesis and biological diversity, which requires excellent preclinical models to allow differential and innovative studies.

Innovative 3D culture methods such as organoid culture have been increasingly implemented in routine in vitro research for many tumor entities. However, due to its distinct biology, organoid culture of HCC has been a challenge. We here describe the initial establishment and characterization of HCC organoid culture.

Materials/Methods:

Surgical HCC tissue specimens were obtained in the operating rooms of the University Hospital Dresden; biopsy specimens were obtained via CT-/ultrasound-guided biopsy of HCC lesions. All samples were processed according to a novel protocol and cultured in Matrigel supplemented with a highly specialized culture medium. Organoids were propagated, passaged and frozen at regular intervals. Proliferation was measured using a modified WST proliferation assay protocol. Next-generation sequencing (NGS) was performed on corresponding triplets of healthy tissue, tumor tissue and organoid samples using the TruSight One Sequencing Panel (Illumina) on an Illumina MiSeq Sequencer. Histology was performed on formalin-fixed and paraffin-embedded organoids using standard protocols.

Results:

We were able to establish a method to culture HCC samples as organoids in vitro. Proliferation assays revealed distinct differences in organoid growth of HCCs from different patients and correlated well with both clinical behavior and histological grading of the tumors. Amplicon sequencing (targeting 4,813 cancer-associated genes) of corresponding sets of tumor specimens and HCC organoids demonstrated the HCC origin of the organoids and their genetic and genomic stability in culture. Treatment of HCC organoids with sorafenib showed the expected reduction in proliferation and viability. Interestingly, treatment of the HCC organoids with hepatocyte growth factor (HGF) resulted in a marked reduction in proliferation as opposed to healthy liver organoids, which had increased proliferation activity upon stimulation with HGF. Histology of the HCC organoids showed distinct features of HCC and corresponded well with the corresponding HCC tissue samples.

Conclusion:

We successfully established a protocol reliably allowing 3D organoid culture of HCC. The organoids faithfully recapitulate the biology of the corresponding tumor and can be used for multiple purposes including in vitro therapeutic testing.

Citation Format: Sebastian Schölch, Lahiri K. Nanduri, Daniel Kühn, Jürgen Weitz, Moritz Koch, Nuh N. Rahbari. Establishment and initial characterization of human hepatocellular carcinoma organoid culture. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4263.