Abstract：
Rat model of N-nitrosomethylbenzylamine (NMBzA)-induced esophageal squamous cell carcinoma (ESCC) is routinely used to study ESCC initiation, progression and new therapeutic strategies. However, the model is time-consuming and malignant tumor incidences are low. Here, we report the usage of multi-kinase inhibitor sorafenib as a tumor promoter to establish an efficient two-stage NMBzA-induced rat ESCC carcinogenesis model, resulting in increments of tumor incidences and shortened tumor formation times. By establishing the model and applying whole-genome sequencing, we discover that benign papillomas and malignant ESCCs harbor most of the \"driver\" events found in rat ESCCs (e.g. recurrent mutations in Ras family, the Hippo and Notch pathways and histone modifier genes) and the mutational landscapes of rat and human ESCCs overlap extensively. We generate tumor cell lines derived from NMBzA-induced papillomas and ESCCs, showing that papilloma cells retain more characteristics of normal epithelial cells than carcinoma cells, especially their exhibitions of normal rat cell karyotypes and inabilities of forming tumors in immunodeficient mice. Three-dimensional (3-D) organoid cultures and single cell RNA sequencing (scRNA-seq) indicate that, when compared to control- and papilloma-organoids, ESCC-organoids display salient abnormalities at tissue and single-cell levels. Multi-omic analyses indicate that NMBzA-induced rat ESCCs are accompanied by progressive hyperactivations of the FAT-Hippo-YAP1 axis and siRNA or inhibitors of YAP1 block the growth of rat ESCCs. Taken together, these studies provide a framework of using an effective rat ESCC model to investigate multilevel functional genomics of ESCC carcinogenesis, which justify targeting YAP1 as a therapeutic strategy for ESCC.
摘要：
N-亚硝基甲基苄胺（NMBzA）诱导的食管鳞状细胞癌（ESCC）的大鼠模型通常用于研究ESCC的发生，进展和新的治疗策略。然而,该模型耗时且恶性肿瘤发病率低。这里,我们报道了使用多激酶抑制剂索拉非尼作为肿瘤促进剂来建立有效的两阶段NMBzA诱导的大鼠ESCC癌变模型，导致肿瘤发病率增加并缩短肿瘤形成时间。通过建立模型并应用全基因组测序,我们发现良性乳头状瘤和恶性ESCC中存在大多数在大鼠ESCC中发现的“驱动”事件(例如Ras家族的复发性突变，Hippo和Notch途径以及组蛋白修饰基因）以及大鼠和人类ESCC的突变景观广泛重叠。我们产生来自NMBzA诱导的乳头状瘤和ESCCs的肿瘤细胞系，表明乳头状瘤细胞比癌细胞保留更多的正常上皮细胞的特征，特别是他们的正常大鼠细胞核型和免疫缺陷小鼠无法形成肿瘤的表现。三维（3-D）类器官培养和单细胞RNA测序（scRNA-seq）表明，与对照和乳头状瘤器官相比，ESCC类器官在组织和单细胞水平显示出显著的异常。多组分析表明，NMBzA诱导的大鼠ESCC伴随着FAT-Hippo-YAP1轴的进行性过度激活，而YAP1的siRNA或抑制剂阻断了大鼠ESCC的生长。一起来看,这些研究提供了使用有效的大鼠ESCC模型来研究ESCC癌变的多级功能基因组学的框架，这证明靶向YAP1作为ESCC的治疗策略是合理的。
