Modeling Wnt signaling by CRISPR-Cas9 genome editing recapitulates neoplasia in human Barrett epithelial organoids.
Modeling Wnt signaling by CRISPR-Cas9 genome editing recapitulates neoplasia in human Barrett epithelial organoids.
复制标题
用CRISPR-Cas9基因组编辑模拟Wnt信号重现人类Barrett上皮器样体中的肿瘤。
DOI：
10.1016/j.canlet.2018.08.017
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发表时间：
2018-11-01
期刊：
影响因子：
9.7
通讯作者：
Meltzer SJ
中科院分区：
文献类型：
--
作者：
Liu X;Cheng Y;Abraham JM;Wang Z;Wang Z;Ke X;Yan R;Shin EJ;Ngamruengphong S;Khashab MA;Zhang G;McNamara G;Ewald AJ;Lin D;Liu Z;Meltzer SJ
由患者活检产生的主要细胞培养物是一个新的疾病模型平台，在体内概括了Barrett食管（BE）。 ies野生型和配对的APC敲除（APCKO）是由CRISPR-CAS9编辑的基因组，表现出特征性的杯状细胞分化。 iferator和复制活动，减少凋亡和延长的文化能力。发挥作用，避免信号传导在永生或癌症衍生的细胞系中呈现的噪声。
Primary organoid cultures generated from patient biopsies comprise a novel improved platform for disease modeling, being genetically stable and closely recapitulating in vivo scenarios. Barrett esophagus (BE) is the major risk factor for esophageal adenocarcinoma. There has been a dearth of long-term in vitro expansion models of BE neoplastic transformation. We generated a long-term virus-free organoid expansion model of BE neoplasia from patient biopsies. Both wild-type and paired APC-knockout (APCKO) BE organoids genome-edited by CRISPR-Cas9 showed characteristic goblet cell differentiation. Autonomous Wnt activation was confirmed in APCKO organoids by overexpression of Wnt target genes and nuclear-translocated β-catenin expression after withdrawal of Wnt-3A and R-spondin-1. Wnt-activated organoids demonstrated histologic atypia, higher proliferative and replicative activity, reduced apoptosis, and prolonged culturability. Wnt-activated organoids also showed sustained protrusive migration ability accompanied by disrupted basement membrane reorganization and integrity. This CRISPR-Cas9 editing human-derived organoid model recapitulates the critical role of aberrant Wnt/β-catenin signaling activation in BE neoplastic transformation. This system can be used to study other ‘driver’ pathway alterations in BE-associated neoplasia, avoiding signaling noise presented in immortalized or cancer-derived cell lines.
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DOI：
10.1126/science.aao3130
发表时间：
2017-10-13
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Drost J;van Boxtel R;Blokzijl F;Mizutani T;Sasaki N;Sasselli V;de Ligt J;Behjati S;Grolleman JE;van Wezel T;Nik-Zainal S;Kuiper RP;Cuppen E;Clevers H
通讯作者：
Clevers H
影响因子：
8
作者：
Clement, G.;Braunschweig, R.;Benhattar, J.
通讯作者：
Benhattar, J.
影响因子：
64.5
作者：
Hsu PD;Lander ES;Zhang F
通讯作者：
Zhang F
影响因子：
82.9
作者：
Huang, Ling;Holtzinger, Audrey;Muthuswamy, Senthil K.
通讯作者：
Muthuswamy, Senthil K.
影响因子：
64.8
作者：
Koo, Bon-Kyoung;Spit, Maureen;Clevers, Hans
通讯作者：
Clevers, Hans