Antibody and Vaccine Group, Centre for Cancer Immunology, University of Southampton Faculty of Medicine, Southampton General Hospital (MP127), Tremona Road, Southampton, Hampshire, SO16 6YD, UK.
Edinburgh Cancer Research Centre, Institute of Genetics and Molecular Medicine, University of Edinburgh, Western General Hospital, Edinburgh, EH4 2XU, UK.
Sci Rep. 2020 Oct 7;10(1):16695. doi: 10.1038/s41598-020-73695-9.
Immunotherapy offers a potentially less toxic, more tumor-specific treatment for neuroblastoma than conventional cytotoxic therapies. Accurate and reproducible immune competent preclinical models are key to understanding mechanisms of action, interactions with other therapies and mechanisms of resistance to immunotherapy. Here we characterized the tumor and splenic microenvironment of two syngeneic subcutaneous (NXS2 and 9464D), and a spontaneous transgenic (TH-MYCN) murine model of neuroblastoma, comparing histological features and immune infiltrates to previously published data on human neuroblastoma. Histological sections of frozen tissues were stained by immunohistochemistry and immunofluorescence for immune cell markers and tumor architecture. Tissues were dissociated by enzymatic digestion, stained with panels of antibodies to detect and quantify cancer cells, along with lymphocytic and myeloid infiltration by flow cytometry. Finally, we tested TH-MYCN mice as a feasible model for immunotherapy, using prior treatment with cyclophosphamide to create a therapeutic window of minimal residual disease to favor host immune development. Immune infiltration differed significantly between all the models. TH-MYCN tumors were found to resemble immune infiltration in human tumors more closely than the subcutaneous models, alongside similar GD2 and MHC class I expression. Finally, TH-MYCN transgenic mice were administered cyclophosphamide alone or in combination with an anti-GD2 or anti-4-1BB monoclonal antibody, which resulted in increase in survival in both combination therapies. The TH-MYCN transgenic mouse is a promising in vivo model for testing immunotherapy compounds and combination therapy in a preclinical setting.
免疫疗法为神经母细胞瘤提供了一种潜在毒性更小、更具肿瘤特异性的治疗方法，优于传统的细胞毒性疗法。准确且可重复的免疫能力临床前模型是理解作用机制、与其他疗法相互作用以及对免疫疗法产生耐药性机制的关键。在这里，我们对两种同基因皮下（NXS2 和 9464D）和一种自发转基因（TH-MYCN）神经母细胞瘤小鼠模型的肿瘤和脾脏微环境进行了特征描述，将组织学特征和免疫浸润与先前发表的人类神经母细胞瘤数据进行了比较。冷冻组织的组织学切片通过免疫组织化学和免疫荧光染色用于免疫细胞标志物和肿瘤结构的检测。通过酶消化使组织解离，并用一系列抗体对其进行染色，以通过流式细胞术检测和量化癌细胞以及淋巴细胞和髓样细胞的浸润。最后，我们使用环磷酰胺预先治疗来创建最小残留疾病的治疗窗口，以有利于宿主免疫发育，从而将 TH-MYCN 小鼠用作免疫疗法的可行模型进行了测试。所有模型之间的免疫浸润差异均有显著差异。与皮下模型相比，TH-MYCN 肿瘤与人类肿瘤的免疫浸润更相似，同时还具有相似的 GD2 和 MHC 类 I 表达。最后，单独或联合使用抗-GD2 或抗-4-1BB 单克隆抗体对 TH-MYCN 转基因小鼠进行了给药，这两种组合疗法均导致了生存率的提高。TH-MYCN 转基因小鼠是一种很有前途的体内模型，可用于在临床前环境中测试免疫疗法化合物和联合疗法。
