Esophageal Adenocarcinoma–Derived Extracellular Vesicle MicroRNAs Induce a Neoplastic Phenotype in Gastric Organoids

• DOI: 10.1016/j.neo.2017.06.007
• Publication Date: 2017-09-30T06:01:24Z
• Authors: Xiquan Ke; Rong Yan; Zhenguo Sun; Yulan Cheng; Amy Meltzer; Nonghua Lu; Xu Shu; Zhe Wang; Binbin Huang; Xi Liu; Zhixiong Wang; Jee Hoon Song; Christopher K Ng; Sariat Ibrahim; John M. Abraham; Eun Ji Shin; Shuixiang He; Stephen J. Meltzer
• Keywords: 0301 basic medicine; Original article; Esophageal Neoplasms; Cell Survival; Neoplasms. Tumors. Oncology. Including cancer and carcinogens; Adenocarcinoma; Coculture Techniques; 3. Good health; Organoids; Extracellular Vesicles; MicroRNAs; 03 medical and health sciences; Cell Transformation, Neoplastic; Phenotype; Gastric Mucosa; Cell Line, Tumor; Humans; RC254-282

There have been no reports describing the effects of cancer cell-derived extracellular vesicles (EVs) on three-dimensional organoids. In this study, we delineated the proneoplastic effects of esophageal adenocarcinoma (EAC)-derived EVs on gastric organoids (gastroids) and elucidated molecular mechanisms underlying these effects. EVs were identified using PKH-67 staining. Morphologic changes, Ki-67 immunochemistry, cell viability, growth rates, and expression levels of miR-25 and miR-210, as well as of their target mRNAs, were determined in gastroids co-cultured with EAC-derived extracellular vesicles (c-EVs). C-EVs were efficiently taken up by gastroids. Notably, c-EV-treated gastroids were more crowded, compact, and multilayered and contained smaller lumens than did those cultured in organoid medium alone or in EAC-conditioned medium that had been depleted of EVs. Moreover, c-EV-treated gastroids manifested increased proliferation and cellular viability relative to medium-only or EV-depleted controls. Expression levels of miR-25 and miR-210 were significantly higher, and those of PTEN and AIFM3 significantly lower, in c-EV-treated versus medium-only or EV-depleted control groups. Inhibitors of miR-25 and miR-210 reversed the increased cell proliferation induced by c-exosomes in co-cultured gastroids by lowering miR-25 and miR-210 levels. In conclusion, we have constructed a novel model system featuring the co-culture of c-EVs with three-dimensional gastroids. Using this model, we discovered that cancer-derived EVs induce a neoplastic phenotype in gastroids. These changes are due, at least in part, to EV transfer of miR-25 and miR-210.