Division of Genetics, Cancer Research Institute, Kanazawa University, Kanazawa, Japan.
Nano Life Science Institute (WPI-NanoLSI), Kanazawa University, Kanazawa, Japan.
Methods Mol Biol. 2024;2828:57-68. doi: 10.1007/978-1-0716-4023-4_6.
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.
最近的癌症基因组分析已经确定了经常发生突变的基因，这些基因是癌症（包括结直肠癌）发生和恶性进展的原因。我们之前构建了携带结直肠癌主要驱动突变的小鼠模型，即 Apc、Kras、Tgfbr2、Trp53 和 Fbxw7 的组合。这些模型的全面组织学分析表明，突变组合与侵袭、上皮-间充质转化（EMT）和转移等恶性表型之间存在关联。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，我们已经从不同基因型的小鼠中建立了肠道肿瘤衍生的类器官，并通过将类器官移植到脾脏中来生成肝转移模型。通过对移植模型的组织学和成像分析，我们确定 Apc、Kras、Tgfbr2 和 Trp53 突变的组合以高发生率促进肝转移。我们还通过我们的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的转移，构成了一种用于基础和临床癌症研究的有用工具，作为临床前模型。我们在此报告类器官培养和转移模型生成的实验方案。
