Department of Pathology, The First Affiliated Hospital of Xi' an Jiaotong University, No. 277 Yanta West Road, Xi' an, 710061, Shaanxi, China; Division of Gastroenterology, Department of Medicine, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, School of Medicine, Baltimore, MD, 21205, USA; Division of Gastroenterology, Department of Oncology, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, School of Medicine, Baltimore, MD, 21205, USA.
Division of Gastroenterology, Department of Medicine, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, School of Medicine, Baltimore, MD, 21205, USA; Division of Gastroenterology, Department of Oncology, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, School of Medicine, Baltimore, MD, 21205, USA.
Cancer Lett. 2018 Nov 1;436:109-118. doi: 10.1016/j.canlet.2018.08.017. Epub 2018 Aug 23.
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 (APC) BE organoids genome-edited by CRISPR-Cas9 showed characteristic goblet cell differentiation. Autonomous Wnt activation was confirmed in APC 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 present in immortalized or cancer-derived cell lines.
从患者活检样本中生成的主要类器官培养物构成了一种新颖的改进的疾病建模平台，具有遗传稳定性，并紧密再现体内情况。巴雷特食管（BE）是食管腺癌的主要危险因素。目前缺乏 BE 肿瘤转化的长期体外扩展模型。我们从患者活检样本中生成了一种长期无病毒 BE 肿瘤类器官扩展模型。通过 CRISPR-Cas9 进行基因组编辑的野生型和配对 APC 敲除（APC）BE 类器官均显示出特征性的杯状细胞分化。在 APC 类器官中，通过 Wnt 靶基因的过表达和 Wnt-3A 和 R- 分泌蛋白 1 撤去后核转位的 β-连环蛋白表达，证实了自主 Wnt 激活。Wnt 激活的类器官表现出组织学异型性、更高的增殖和复制活性、减少的细胞凋亡和延长的可培养性。Wnt 激活的类器官还显示出持续的伸出迁移能力，伴随着基底膜重组和完整性的破坏。这种 CRISPR-Cas9 编辑的人源类器官模型再现了异常 Wnt/β-连环蛋白信号激活在 BE 肿瘤转化中的关键作用。该系统可用于研究 BE 相关肿瘤中其他“驱动”途径的改变，避免了在永生化或癌症衍生细胞系中存在的信号噪声。
