Sugahara Osamu, Koga Daisuke, Oka Takeru, Sugiyama Shigeaki, Wada Reona, Higa Tsunaki, Nakayama Keiichi I
Anticancer Strategies Laboratory, Advanced Research Initiative, Institute of Science Tokyo, Tokyo, Japan.
Department of Molecular and Cellular Biology, Medical Institute of Bioregulation, Kyushu University, Fukuoka, Japan.
Cancer Sci. 2025 Sep;116(9):2457-2470. doi: 10.1111/cas.70129. Epub 2025 Jul 7.
The tumor immune microenvironment plays a key role in the regulation of cancer progression. Recent studies have suggested a relation between diverse tumor genotypes and tumor immune microenvironment phenotypes for cholangiocarcinoma (CCA). However, the contribution of tumor-infiltrating immune cells to CCA progression has remained unclear, underscoring the need for genetically defined CCA models in immunocompetent mice. We here aimed to generate genetically engineered and transplantable CCA organoids from C57BL/6 mice and to investigate the role of tumor-infiltrating immune cells in CCA progression with this model. CCA organoids were generated ex vivo with the use of the CRISPR/Cas9 system. Orthotopic transplantation of CCA organoids harboring mutations in Smad4, Trp53, and Kras into wild-type C57BL/6 mice resulted in tumor formation accompanied by distant metastasis. Selective depletion of immune cell types in the tumor-bearing mice revealed an antitumor action of tumor-infiltrating neutrophils (TINs) that was mediated by direct killing of cancer cells through the production of reactive oxygen species. Furthermore, administration of recombinant human granulocyte colony-stimulating factor (rhG-CSF) increased the number and cytotoxicity of TINs, suppressed tumor growth, and prolonged the survival of tumor-bearing mice. Finally, combination treatment with rhG-CSF and standard chemotherapy resulted in a synergistic attenuation of tumor growth. Our study therefore provides a syngeneic and genetically defined mouse model of CCA and highlights the therapeutic potential of targeting TINs with rhG-CSF.
肿瘤免疫微环境在癌症进展的调控中起关键作用。最近的研究表明，胆管癌（CCA）的多种肿瘤基因型与肿瘤免疫微环境表型之间存在关联。然而，肿瘤浸润免疫细胞对CCA进展的作用仍不清楚，这突出了在具有免疫活性的小鼠中建立基因定义的CCA模型的必要性。我们的目的是从C57BL/6小鼠中生成基因工程化且可移植的CCA类器官，并利用该模型研究肿瘤浸润免疫细胞在CCA进展中的作用。使用CRISPR/Cas9系统在体外生成CCA类器官。将在Smad4、Trp53和Kras中发生突变的CCA类器官原位移植到野生型C57BL/6小鼠中，导致肿瘤形成并伴有远处转移。对荷瘤小鼠的免疫细胞类型进行选择性清除，揭示了肿瘤浸润中性粒细胞（TINs）的抗肿瘤作用，该作用是通过产生活性氧直接杀伤癌细胞介导的。此外，给予重组人粒细胞集落刺激因子（rhG-CSF）可增加TINs的数量和细胞毒性，抑制肿瘤生长，并延长荷瘤小鼠的生存期。最后，rhG-CSF与标准化疗联合治疗导致肿瘤生长的协同减弱。因此，我们的研究提供了一种同基因且基因定义的CCA小鼠模型，并突出了用rhG-CSF靶向TINs的治疗潜力。
