Takei Jun, Furudate Ken, Nagaoka-Kamata Yoshiko, Iwaloye Opeyemi, Jepson Chloe E, Blucas Madison T, Saito Kiyotaka, Welner Robert S, Van Meir Erwin G, Kamata Masakazu, Osuka Satoru
Department of Neurosurgery, Heersink School of Medicine, University of Alabama at Birmingham, Birmingham, Alabama, USA.
Department of Oral and Maxillofacial Surgery, Hirosaki University Graduate School of Medicine, Hirosaki, Japan.
bioRxiv. 2025 May 22:2025.05.17.654526. doi: 10.1101/2025.05.17.654526.
Glioblastoma (GBM) is the deadliest primary brain tumor in adults, where current therapies fail to meaningfully extend survival. Available animal GBM tumor models, especially therapy-resistant and recurrent ones with unique immunological aspects, are restricted, impeding innovative treatment research. To confront this critical obstacle we established a unique GBM mouse model that utilizes patient-derived xenografts (PDXs) within humanized mice.
We selected two immune-deficient mouse models to facilitate the reconstitution of myeloid lineage cells. After undergoing myeloablation, mice received CD34+ hematopoietic stem progenitor cells derived from human umbilical cord blood for humanization. Upon confirming the reconstitution of human blood cells, mice were xenografted with PDXs resistant to radiation. Tumor profiles and immune cell infiltration were analyzed via flow cytometry, immunohistochemistry, and single-cell RNA sequencing (scRNA-seq). The findings were evaluated against scRNA-seq data from recurrent human GBM.
A diverse range of human immune cells, including T, NK, and myeloid lineage cells, infiltrated PDX tumors in humanized mice. Notably, gene expression profiles in these immune cells resembled that of recurrent human GBM. Unlike conventional xenograft models, this model highlighted enhanced tumor diversity, particularly a high fraction of neural progenitor-like cells.
Our humanized GBM mouse model displayed an immune cell signature similar to recurrent GBM. This model is a valuable resource for analyzing the tumor immune landscape and assessing new therapies, particularly immunotherapies. By enabling effective evaluation of novel treatments, our model has the potential to significantly advance GBM research.
胶质母细胞瘤（GBM）是成人中最致命的原发性脑肿瘤，目前的治疗方法未能显著延长生存期。现有的动物GBM肿瘤模型，尤其是具有独特免疫特征的治疗抵抗性和复发性模型有限，阻碍了创新性治疗研究。为了克服这一关键障碍，我们建立了一种独特的GBM小鼠模型，该模型在人源化小鼠中利用患者来源的异种移植（PDX）。
我们选择了两种免疫缺陷小鼠模型来促进髓系细胞的重建。在进行清髓后，小鼠接受源自人脐带血的CD34+造血干祖细胞进行人源化。在确认人血细胞重建后，将对辐射耐药的PDX移植到小鼠体内。通过流式细胞术、免疫组织化学和单细胞RNA测序（scRNA-seq）分析肿瘤特征和免疫细胞浸润情况。将研究结果与复发性人类GBM的scRNA-seq数据进行对比评估。
多种人类免疫细胞，包括T细胞、NK细胞和髓系细胞，浸润了人源化小鼠中的PDX肿瘤。值得注意的是，这些免疫细胞中的基因表达谱与复发性人类GBM相似。与传统的异种移植模型不同，该模型突出了肿瘤多样性的增强，特别是高比例的神经祖细胞样细胞。
我们的人源化GBM小鼠模型显示出与复发性GBM相似的免疫细胞特征。该模型是分析肿瘤免疫格局和评估新疗法，特别是免疫疗法的宝贵资源。通过有效评估新疗法，我们的模型有潜力显著推进GBM研究。
