Establishment and drug screening of patient-derived extrahepatic biliary tract carcinoma organoids

Abstract

Background Patient-derived organoids (PDO) have been proposed as a novel in vitro method of drug screening for different types of cancer. However, to date, extrahepatic biliary tract carcinoma (eBTC) PDOs have not yet been fully established. Methods We collected six samples of gallbladder carcinoma (GBC) and one sample of extrahepatic cholangiocarcinoma (eCCA) from seven patients to attempt to establish eBTC PDOs for drug screening. We successfully established five GBC and one eCCA PDOs. Histological staining was used to compare structural features between the original tissues and cancer PDOs. Whole exome sequencing (WES) was performed to analyze the genetic profiles of original tissues and cancer PDOs. Drug screening, including gemcitabine, 5-fluorouracil, cisplatin, paclitaxel, infigratinib, and ivosidenib, was measured and verified by clinical effects in certain cases. Results Different PDOs exhibited diverse growth rates during in vitro culture. Hematoxylin and eosin staining demonstrated that the structures of most cancer PDOs retained the original structures of adenocarcinoma. Immunohistological and periodic acid-schiff staining revealed that marker expression in cancer PDOs was similar to that of the original specimens. Genetic profiles from the four original specimens, as well as paired cancer PDOs, were analyzed using whole exome sequencing. Three of the four PDOs exhibited a high degree of similarity when compared to the original specimens, except for GBC2 PDO, which only had a concordance of 74% in the proportion of single nucleotide polymorphisms in the coding sequence. In general, gemcitabine was found to be the most efficient drug for eBTC treatment, as it showed moderate or significant inhibitory impact on cancer growth. Results from drug screening were confirmed to a certain extent by three clinical cases. Conclusions Our study successfully established a series of eBTC PDOs, which contributed to the field of eBTC PDOs. Additional enhancements should be explored to improve the growth rate of PDOs and to preserve their immune microenvironment.

Advanced Cellular Models for Preclinical Drug Testing: From 2D Cultures to Organ-on-a-Chip Technology

“…As an outcome of this discovery, methods to establish patient-derived tumor organoids (PDTOs) have been developed that can be cultured and expanded in clinically relevant timeframes in the search for personalized therapies [ 49 , 50 , 51 , 52 , 53 ]. Research on PDTOs dates back to one decade ago, when original work on pancreatic cancer has shown the setup of culture conditions for primary pancreatic ductal adenocarcinoma (PDAC) organoids that retained in vitro the same tissue architecture and cell differentiation of the tumor in vivo.…”

Organoids: The current status and biomedical applications

“…Thus, organoid models provide better options for drug screening and individualized drug therapy. [29][30][31][32][33] The potential of organoids to broaden fundamental research by complementing current model systems is becoming more generally recognized. 34 Organoid technologies pave the way for the development of new models that more closely resemble realistic physiological and pathophysiological states.…”

“…Tumor organoids can be constructed by preoperative biopsy or postoperative tumor resection, which plays an important role in individualized drug sensitivity prediction. Thus, organoid models provide better options for drug screening and individualized drug therapy 29–33 . The potential of organoids to broaden fundamental research by complementing current model systems is becoming more generally recognized 34 …”

Lactate promotes the growth of patient-derived organoids from hepatopancreatobiliary cancers via ENO1/HIF1α pathway and does not affect their drug sensitivities

“…PDOs of HCC, PDC, CC, colorectal cancer, breast cancer, gastric cancer and bladder cancer have been established so far [ 11 – 17 ]. Recently, we established PDOs from intrahepatic and extrahepatic CC tissues and verified the consistency between the drug sensitivity of these organoids and the actual efficacy of anticancer treatment [ 18 , 19 ]. However, although most PDOs can be constructed within several weeks, those derived from the stromal cell-rich adenocarcinomas still require several months.…”

“…Given the low 5-year survival rates of patients with advanced CC or PDC, it is imperative to obtain PDOs within a reasonable time frame. We cultured PDOs in previous studies for no more than two weeks and thus only a few drugs were used in drug screening [ 18 , 19 ]. Yin et al analyzed the genetic profiles of several PDOs by mRNA sequencing and accordingly cultured them with suitable supplements [ 21 ].…”