最近的癌症基因组分析发现了经常突变的基因，这些基因导致癌症（包括结直肠癌（CRC））的发生和恶性进展。我们之前构建了携带 CRC 主要驱动突变的小鼠模型，即 Apc、Kras、Tgfbr2、Trp53 和 Fbxw7 的组合。对模型的综合组织学分析显示突变组合与恶性表型（例如侵袭、上皮间质转化（EMT）和转移）之间存在联系。癌症相关死亡的主要原因是转移，因此了解转移的机制对于开发新的治疗策略非常重要。为此，我们建立了来自不同基因型小鼠的肠道肿瘤来源的类器官，并通过将类器官移植到脾脏中生成肝转移模型。通过对移植模型的组织学和影像学分析，我们确定Apc、Kras、Tgfbr2和Trp53突变的组合促进肝转移的高发生率。我们还通过模型分析证明了由遗传和表型不同的细胞组成的肿瘤细胞簇的多克隆转移。这些类器官移植模型再现了人类结直肠癌转移，作为临床前模型构成了基础和临床癌症研究的有用工具。我们在此报告类器官培养和转移模型生成的实验方案。© 2024。作者获得 Springer Science Business Media, LLC（Springer Nature 的一部分）的独家许可。

Recent cancer genome analyses have identified frequently mutated genes that are responsible for the development and malignant progression of cancers, including colorectal cancer (CRC). We previously constructed mouse models that carried major driver mutations of CRC, namely Apc, Kras, Tgfbr2, Trp53, and Fbxw7, in combinations. Comprehensive histological analyses of the models showed a link between mutation combinations and malignant phenotypes, such as invasion, epithelial-mesenchymal transition (EMT), and metastasis. The major cause of cancer-related death is metastasis, making it important to understand the mechanism underlying metastasis in order to develop novel therapeutic strategies. To this end, we have established intestinal tumor-derived organoids from different genotyped mice and generated liver metastasis models via transplantation of the organoids into the spleen. Through histological and imaging analyses of the transplantation models, we have determined that the combination of Apc, Kras, Tgfbr2, and Trp53 mutations promotes liver metastasis at a high incidence. We also demonstrated polyclonal metastasis of tumor cell clusters consisting of genetically and phenotypically distinct cells through our model analysis. These organoid transplantation models recapitulate human CRC metastasis, constituting a useful tool for basic and clinical cancer research as a preclinical model. We herein report the experimental protocols of the organoid culture and generation of metastasis models.© 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.