An Organoid Biobank of Neuroendocrine Neoplasms Enables Genotype-Phenotype Mapping
Kenta Kawasaki, Kohta Toshimitsu, Mami Matano, Masashi Fujita, Masayuki Fujii, Kazuhiro Togasaki, Toshiki Ebisudani, Mariko Shimokawa, Ai Takano, Sirirat Takahashi, Yuki Ohta, Kosaku Nanki, Ryo Igarashi, Kazuhiro Ishimaru, Hiroki Ishida, Yasutaka Sukawa, Shinya Sugimoto, Yoshimasa Saito, Kazuhiro Maejima, Shota Sasagawa, Hwajin Lee, Hong-Gee Kim, Kyungsik Ha, Junko Hamamoto, Koichi Fukunaga, Aya Maekawa, Minoru Tanabe, Soichiro Ishihara, Yasuo Hamamoto, Hiroyuki Yasuda, Shigeki Sekine, Atsushi Kudo, Yuko Kitagawa, Takanori Kanai, Hidewaki Nakagawa, Toshiro Sato, Kenta Kawasaki, Kohta Toshimitsu, Mami Matano, Masashi Fujita, Masayuki Fujii, Kazuhiro Togasaki, Toshiki Ebisudani, Mariko Shimokawa, Ai Takano, Sirirat Takahashi, Yuki Ohta, Kosaku Nanki, Ryo Igarashi, Kazuhiro Ishimaru, Hiroki Ishida, Yasutaka Sukawa, Shinya Sugimoto, Yoshimasa Saito, Kazuhiro Maejima, Shota Sasagawa, Hwajin Lee, Hong-Gee Kim, Kyungsik Ha, Junko Hamamoto, Koichi Fukunaga, Aya Maekawa, Minoru Tanabe, Soichiro Ishihara, Yasuo Hamamoto, Hiroyuki Yasuda, Shigeki Sekine, Atsushi Kudo, Yuko Kitagawa, Takanori Kanai, Hidewaki Nakagawa, Toshiro Sato
Abstract
Gastroenteropancreatic (GEP) neuroendocrine neoplasm (NEN) that consists of neuroendocrine tumor and neuroendocrine carcinoma (NEC) is a lethal but under-investigated disease owing to its rarity. To fill the scarcity of clinically relevant models of GEP-NEN, we here established 25 lines of NEN organoids and performed their comprehensive molecular characterization. GEP-NEN organoids recapitulated pathohistological and functional phenotypes of the original tumors. Whole-genome sequencing revealed frequent genetic alterations in TP53 and RB1 in GEP-NECs, and characteristic chromosome-wide loss of heterozygosity in GEP-NENs. Transcriptome analysis identified molecular subtypes that are distinguished by the expression of distinct transcription factors. GEP-NEN organoids gained independence from the stem cell niche irrespective of genetic mutations. Compound knockout of TP53 and RB1, together with overexpression of key transcription factors, conferred on the normal colonic epithelium phenotypes that are compatible with GEP-NEN biology. Altogether, our study not only provides genetic understanding of GEP-NEN, but also connects its genetics and biological phenotypes.
Keywords: 3D culture; CRISPR-Cas9; NKX2-5; epigenome; fusion gene; gastrinoma; lineage reprogramming; multi-omics; stem cell niche; transcription factors.
Conflict of interest statement
Declaration Of Interests The authors declare no competing interests.
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Source: PubMed