Three-Dimensional Organoids Reveal Therapy Resistance of Esophageal and Oropharyngeal Squamous Cell Carcinoma Cells

• Published in: Cellular and molecular gastroenterology and hepatology Vol. 7; no. 1; pp. 73 - 91
• Main Authors: Kijima, Takashi, Nakagawa, Hiroshi, Shimonosono, Masataka, Chandramouleeswaran, Prasanna M., Hara, Takeo, Sahu, Varun, Kasagi, Yuta, Kikuchi, Osamu, Tanaka, Koji, Giroux, Veronique, Muir, Amanda B., Whelan, Kelly A., Ohashi, Shinya, Naganuma, Seiji, Klein-Szanto, Andres J., Shinden, Yoshiaki, Sasaki, Ken, Omoto, Itaru, Kita, Yoshiaki, Muto, Manabu, Bass, Adam J., Diehl, J. Alan, Ginsberg, Gregory G., Doki, Yuichiro, Mori, Masaki, Uchikado, Yasuto, Arigami, Takaaki, Avadhani, Narayan G., Basu, Devraj, Rustgi, Anil K., Natsugoe, Shoji
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.01.2019; Elsevier
• Subjects: 3D Organoids; 5-Fluorouracil; Animals; Autophagy; Autophagy - drug effects; Biopsy; Carcinoma, Squamous Cell - pathology; Carcinoma, Squamous Cell - therapy; CD44; Cell Line, Tumor; Chemoradiotherapy; Drug Resistance, Neoplasm; Endoscopy; Esophageal Neoplasms - pathology; Fluorouracil - pharmacology; Fluorouracil - therapeutic use; Gastroenterology and Hepatology; Humans; Hyaluronan Receptors - metabolism; Mice; Organoids - pathology; Original Research; Oropharyngeal Neoplasms - pathology; Oropharyngeal Neoplasms - therapy; 5-Fluorouracil; 5FU; IHC; H&E; SCCs; CD44H; OPSCC; Autophagy; EMT; CQ; ESCC; DMEM; 3D; 3D Organoids; AV; FBS; IC50; PI; LC3; CD44; TE11R; IC 50; half maximal inhibitory concentration; chloroquine; propidium iodide; oropharyngeal squamous cell carcinoma; epithelial-mesenchymal transition; immunohistochemistry; 3-dimensional; hematoxylin and eosin; esophageal squamous cell carcinoma; light chain 3; squamous cell carcinomas; high expression of CD44; autophagy vesicle; fetal bovine serum; Dulbecco’s modified Eagle medium; 5-fluorouracil–resistant derivative of TE11; EMT, epithelial-mesenchymal transition; CD44H, high expression of CD44; PI, propidium iodide; ESCC, esophageal squamous cell carcinoma; FBS, fetal bovine serum; IC50, half maximal inhibitory concentration; LC3, light chain 3; CQ, chloroquine; SCCs, squamous cell carcinomas; TE11R, 5-fluorouracil–resistant derivative of TE11; H&E, hematoxylin and eosin; IHC, immunohistochemistry; OPSCC, oropharyngeal squamous cell carcinoma; 5FU, 5-fluorouracil; DMEM, Dulbecco’s modified Eagle medium; AV, autophagy vesicle; 3D, 3-dimensional
• ISSN: 2352-345X; 2352-345X
• DOI: 10.1016/j.jcmgh.2018.09.003

Oropharyngeal and esophageal squamous cell carcinomas, especially the latter, are a lethal disease, featuring intratumoral cancer cell heterogeneity and therapy resistance. To facilitate cancer therapy in personalized medicine, three-dimensional (3D) organoids may be useful for functional characterization of cancer cells ex vivo. We investigated the feasibility and the utility of patient-derived 3D organoids of esophageal and oropharyngeal squamous cell carcinomas. We generated 3D organoids from paired biopsies representing tumors and adjacent normal mucosa from therapy-naïve patients and cell lines. We evaluated growth and structures of 3D organoids treated with 5-fluorouracil ex vivo. Tumor-derived 3D organoids were grown successfully from 15 out of 21 patients (71.4%) and passaged with recapitulation of the histopathology of the original tumors. Successful formation of tumor-derived 3D organoids was associated significantly with poor response to presurgical neoadjuvant chemotherapy or chemoradiation therapy in informative patients (P = 0.0357, progressive and stable diseases, n = 10 vs. partial response, n = 6). The 3D organoid formation capability and 5-fluorouracil resistance were accounted for by cancer cells with high CD44 expression and autophagy, respectively. Such cancer cells were found to be enriched in patient-derived 3D organoids surviving 5-fluorouracil treatment. The single cell-based 3D organoid system may serve as a highly efficient platform to explore cancer therapeutics and therapy resistance mechanisms in conjunction with morphological and functional assays with implications for translation in personalized medicine. [Display omitted]