Patient-Derived Organoids of Cholangiocarcinoma

Abstract

Cholangiocarcinoma (CC) is an aggressive malignancy with an inferior prognosis due to limited systemic treatment options. As preclinical models such as CC cell lines are extremely rare, this manuscript reports a protocol of cholangiocarcinoma patient-derived organoid culture as well as a protocol for the transition of 3D organoid lines to 2D cell lines. Tissue samples of non-cancer bile duct and cholangiocarcinoma were obtained during surgical resection. Organoid lines were generated following a standardized protocol. 2D cell lines were generated from established organoid lines following a novel protocol. Subcutaneous and orthotopic patient-derived xenografts were generated from CC organoid lines, histologically examined, and treated using standard CC protocols. Therapeutic responses of organoids and 2D cell lines were examined using standard CC agents. Next-generation exome and RNA sequencing was performed on primary tumors and CC organoid lines. Patient-derived organoids closely recapitulated the original features of the primary tumors on multiple levels. Treatment experiments demonstrated that patient-derived organoids of cholangiocarcinoma and organoid-derived xenografts can be used for the evaluation of novel treatments and may therefore be used in personalized oncology approaches. In summary, this study establishes cholangiocarcinoma organoids and organoid-derived cell lines, thus expanding translational research resources of cholangiocarcinoma.

Application of Patient-Derived Cancer Organoids to Personalized Medicine

“…There are some reports of screening only FDA-approved drugs or clinically available agents for treatment because it is impractical to screen compounds that cannot be used clinically [ 58 , 74 , 75 , 76 , 77 ]. Ramamoorthy et al developed a multicellular lung organoid called primitive lung-in-a-dish (PLiD), which mimics the lung microstructure with air sac-like alveoli and produces lung surfactant protein.…”

Organoid Models for Cancer Research—From Bed to Bench Side and Back

“…In 2011, Sato et al first established an organoid culture system of benign and malign PDOs of the colon and esophagus [ 30 ]. These PDOs resemble the individual histopathological features, tumor stage, genetic landscape as well as cellular heterogeneity of the original tumor of the individual patient [ 54 , 55 , 56 , 57 , 58 , 59 ]. This applies not only for primary tumors but has also been shown for organoids derived from metastases or even from extremely rare circulating tumor cells (CTCs) [ 60 , 61 , 62 ].…”

Integrative analysis of multiple genomic data from intrahepatic cholangiocarcinoma organoids enables tumor subtyping

“…Recently developed organoid culture techniques enable formation of adult stem cell-derived cancer organoids from cholangiocarcinoma specimens. [12][13][14][15][16][17] Importantly, the resulting organoids show similar phenotypic and genetic characteristics compared with the original primary tumor. A previous study showed that even small biopsy samples are su cient for the generation of tissue-derived organoids.…”