Department of Neurological Surgery, Northwestern University, Chicago, IL 60611, USA.
Rush Medical College, Rush University Medical Center, Chicago, IL 60612, USA.
Cells. 2023 Jun 25;12(13):1716. doi: 10.3390/cells12131716.
Glioblastoma (GBM) is a lethal brain tumor with limited therapeutic options. Bi-specific killer cell engagers (BiKEs) are novel immunotherapies designed to engage natural killer (NK) cells against cancer. We designed a BiKE molecule consisting of a single-domain CD16 antibody, an interleukin-15 linker, and a single-chain variable antibody against the glioma-associated antigen interleukin 13 receptor alpha 2 (IL13Rα2). Recombinant BiKE protein was expressed in HEK cells and purified. Flow cytometric analysis of co-cultures of peripheral blood-derived NK cells with GBM6 and GBM39 patient-derived xenograft lines revealed significantly increased activation of NK cells (CD25+CD69+) and increased glioma cell killing following BiKE treatment compared to controls (n = 4, < 0.01). Glioma cell killing was also confirmed via immunofluorescence staining for cleaved caspase-3 ( < 0.05). In vivo, intracranial delivery of NK cells with BiKE extended median survival in mice bearing GBM6 ( < 0.01) and GBM12 ( < 0.01) tumors compared to controls. Finally, histological analysis of brain tissues revealed a higher frequency of peritumoral NK cells in mice treated with BiKE than with NK cells alone ( < 0.05). In conclusion, we demonstrate that a BiKE generated in a mammalian expression system is functional in augmenting NK cell targeting of IL13Rα2-positive gliomas.
胶质母细胞瘤（GBM）是一种致命的脑肿瘤，治疗选择有限。双特异性杀伤细胞衔接器（BiKEs）是一种新型免疫疗法，旨在利用自然杀伤（NK）细胞靶向癌症。我们设计了一种由单域 CD16 抗体、白细胞介素 15 接头和针对胶质瘤相关抗原白细胞介素 13 受体 alpha 2（IL13Rα2）的单链可变抗体组成的 BiKE 分子。重组 BiKE 蛋白在 HEK 细胞中表达并纯化。将外周血来源的 NK 细胞与 GBM6 和 GBM39 患者来源的异种移植系共培养的流式细胞术分析显示，与对照组相比，BiKE 治疗后 NK 细胞（CD25+CD69+）的激活显著增加，并且对神经胶质瘤细胞的杀伤增加（n = 4，< 0.01）。通过免疫荧光染色检测到的 cleaved caspase-3 也证实了神经胶质瘤细胞的杀伤（< 0.05）。在体内，与对照组相比，携带 BiKE 的 NK 细胞颅内递送延长了携带 GBM6（< 0.01）和 GBM12（< 0.01）肿瘤的小鼠的中位生存期。最后，脑组织的组织学分析显示，与单独使用 NK 细胞相比，用 BiKE 治疗的小鼠中肿瘤周围的 NK 细胞频率更高（< 0.05）。总之，我们证明了在哺乳动物表达系统中产生的 BiKE 可增强 NK 细胞对 IL13Rα2 阳性神经胶质瘤的靶向作用。
