ヒト消化器がん(大腸, 胃, 肝臓, 膵臓)患者から採取した生検組織からがんオルガノイドを樹立し, 最適化した培養環境下において大量培養することに成功した。また遺伝子操作技術を用いて蛍光蛋白レポーターLgr5遺伝子座位へのノックインによりがん幹細胞の可視化に成功し, この技術を利用して, 樹立した消化器がん幹細胞の可視化を行った。これらの樹立したがんオルガノイドを用いてHTSを行った。可視化した消化器がんオルガノイドに対し, 360種類のPKI libraryを用い, HTSを行った結果, 各消化器がん細胞及びがん幹細胞に効果を有する薬剤がいくつか得られた。
We have established gastrointestinal cancer organoids from the patient. As a result, we succeeded in isolating 4 kinds of tumor organoids including colorectal, gastric , liver, and pancreatic cancer from patient. It was possible to continuously and long-term culture of each of the human gastrointestinal cancer organoids by optimizing the culture condition based on the orgnoid culture protocol. We also have been successful knocked-in reporter to LGR5 locus of intestinal organoids using genome editing. Each gastrointestinal cancer organoids derived from patients became culture in 384-well plate. Furthermore, we have established tumor organoids ; based High-throughput drug screen system. Image analysis was performed with 360 drugs on tumor organoids using High Contents Analyzer. Our results suggest that our established HTS for tumor organids will be apply to genetic mutation based drug discovery and personalized medicine.
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