CD133 marks self-renewing cancer stem cells (CSCs) in a variety of solid tumors, and CD133+ tumor-initiating cells are known markers of chemo- and radio-resistance in multiple aggressive cancers, including glioblastoma (GBM), that may drive intra-tumoral heterogeneity. Here, we report three immunotherapeutic modalities based on a human anti-CD133 antibody fragment that targets a unique epitope present in glycosylated and non-glycosylated CD133 and studied their effects on targeting CD133+ cells in patient-derived models of GBM. We generated an immunoglobulin G (IgG) (RW03-IgG), a dual-antigen T cell engager (DATE), and a CD133-specific chimeric antigen receptor T cell (CAR-T): CART133. All three showed activity against patient-derived CD133+ GBM cells, and CART133 cells demonstrated superior efficacy in patient-derived GBM xenograft models without causing adverse effects on normal CD133+ hematopoietic stem cells in humanized CD34+ mice. Thus, CART133 cells may be a therapeutically tractable strategy to target CD133+ CSCs in human GBM or other treatment-resistant primary cancers.

CD133在多种实体瘤中标记自我更新的癌症干细胞（CSC），而CD133 +肿瘤起始细胞是多种侵袭性癌症（包括胶质母细胞瘤（GBM））中化学耐药和放射耐药的已知标志物，可能会导致肿瘤异质性。在这里，我们报告了三种基于人类抗CD133抗体片段的免疫治疗方法，该片段靶向糖基化和非糖基化CD133中存在的独特表位，并研究了它们在GBM患者衍生模型中靶向CD133 +细胞的作用。我们生成了免疫球蛋白G（IgG）（RW03-IgG），双重抗原T细胞衔接子（DATE）和CD133特异性嵌合抗原受体T细胞（CAR-T）：CART133。这三者均表现出针对源自患者的CD133 + GBM细胞的活性，CART133和CART133细胞在源自患者的GBM异种移植模型中显示出优异的功效，而不会对人源化CD34 +小鼠的正常CD133 +造血干细胞产生不利影响。因此，CART133细胞可能是靶向人类GBM或其他具有治疗抵抗力的原发性癌症中CD133 + CSCs的治疗方法。