免疫治疗创新对于局限性可手术肿瘤至关重要。CAR-T细胞疗法对于胶质母细胞瘤（GBM）的效率降低，可能是由于疾病进展下的突变所致。然而，自然杀伤细胞（NK细胞）能够检测到这些突变的癌细胞，展现出较高的肿瘤消除潜力。我们利用人类多能干细胞（hPSCs）建立了一种NK细胞分化系统。通过该系统，我们可以在多潜能阶段引入针对癌症治疗挑战的基因修饰，从而实现修饰的“现成”hPSC-NKs的无限生产。我们采用自主开发的器官样结构体系将hPSCs分化为造血前体细胞（HPCs）和NK细胞。我们使用流式细胞仪和生物信息学分析对这些细胞进行了表征。采用NSG小鼠评估了HPC的移植能力。我们使用体外和体内的K562实验验证了NK细胞的细胞毒性，并在体外进一步证实了对淋巴瘤、弥漫性内生性脑干胶质瘤（DIPG）和GBM细胞系的细胞毒性。HPCs表现出在外周血样本中的移植能力，而hPSC-NKs则展示了与同一供体的外周血NK细胞相似的形态和功能。hPSC-NKs在检查点抑制剂和代谢基因表达方面也显示出潜在优势，并展示了对各种癌症的体外和体内细胞毒性。我们设计了一种能够复制体内细胞组织结构（包括信号梯度和剪切应力条件）的器官样结构体系，为HPC和NK的生成提供了适宜的环境。HPC的可移植性和NK对白血病、淋巴瘤、DIPG和GBM的强大细胞毒性突出了该创新体系的潜力，使其成为一种有价值的工具，有助于癌症治疗和研究，提高患者的生存率和生活质量。 © 2023 The Author(s). Published with license by Taylor & Francis Group, LLC.

Immunotherapeutic innovation is crucial for limited operability tumors. CAR T-cell therapy displayed reduced efficiency against glioblastoma (GBM), likely due to mutations underlying disease progression. Natural Killer cells (NKs) detect cancer cells despite said mutations - demonstrating increased tumor elimination potential. We developed an NK differentiation system using human pluripotent stem cells (hPSCs). Via this system, genetic modifications targeting cancer treatment challenges can be introduced during pluripotency - enabling unlimited production of modified "off-the-shelf" hPSC-NKs.hPSCs were differentiated into hematopoietic progenitor cells (HPCs) and NKs using our novel organoid system. These cells were characterized using flow cytometric and bioinformatic analyses. HPC engraftment potential was assessed using NSG mice. NK cytotoxicity was validated using in vitro and in vitro K562 assays and further corroborated on lymphoma, diffuse intrinsic pontine glioma (DIPG), and GBM cell lines in vitro.HPCs demonstrated engraftment in peripheral blood samples, and hPSC-NKs showcased morphology and functionality akin to same donor peripheral blood NKs (PB-NKs). The hPSC-NKs also displayed potential advantages regarding checkpoint inhibitor and metabolic gene expression, and demonstrated in vitro and in vivo cytotoxicity against various cancers.Our organoid system, designed to replicate in vivo cellular organization (including signaling gradients and shear stress conditions), offers a suitable environment for HPC and NK generation. The engraftable nature of HPCs and potent NK cytotoxicity against leukemia, lymphoma, DIPG, and GBM highlight the potential of this innovative system to serve as a valuable tool that will benefit cancer treatment and research - improving patient survival and quality of life.© 2023 The Author(s). Published with license by Taylor & Francis Group, LLC.