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一种可扩展的、无自旋的方法,用于生成增强的诱导多能干细胞衍生的自然杀伤细胞,用于癌症免疫治疗。

A scalable, spin-free approach to generate enhanced induced pluripotent stem cell-derived natural killer cells for cancer immunotherapy.

机构信息

Frazer Institute, Faculty of Medicine, The University of Queensland, Woolloongabba, QLD, Australia.

Australian Institute for Bioengineering and Nanotechnology, University of Queensland, St Lucia, QLD, Australia.

出版信息

Immunol Cell Biol. 2024 Nov;102(10):924-934. doi: 10.1111/imcb.12820. Epub 2024 Sep 13.


DOI:10.1111/imcb.12820
PMID:39269338
Abstract

Natural killer (NK) cells play a vital role in innate immunity and show great promise in cancer immunotherapy. Traditional sources of NK cells, such as the peripheral blood, are limited by availability and donor variability. In addition, in vitro expansion can lead to functional exhaustion and gene editing challenges. This study aimed to harness induced pluripotent stem cell (iPSC) technology to provide a consistent and scalable source of NK cells, overcoming the limitations of traditional sources and enhancing the potential for cancer immunotherapy applications. We developed human placental-derived iPSC lines using reprogramming techniques. Subsequently, an optimized two-step differentiation protocol was introduced to generate high-purity NK cells. Initially, iPSCs were differentiated into hematopoietic-like stem cells using spin-free embryoid bodies (EBs). Subsequently, the EBs were transferred to ultra-low attachment plates to induce NK cell differentiation. iPSC-derived NK (iNK) cells expressed common NK cell markers (NKp46, NKp30, NKp44, CD16 and eomesodermin) at both RNA and protein levels. iNK cells demonstrated significant resilience to cryopreservation and exhibited enhanced cytotoxicity. The incorporation of a chimeric antigen receptor (CAR) construct further augmented their cytotoxic potential. This study exemplifies the feasibility of generating iNK cells with high purity and enhanced functional capabilities, their improved resilience to cryopreservation and the potential to have augmented cytotoxicity through CAR expression. Our findings offer a promising pathway for the development of potential cellular immunotherapies, highlighting the critical role of iPSC technology in overcoming challenges associated with traditional NK cell sources.

摘要

自然杀伤 (NK) 细胞在先天免疫中发挥着至关重要的作用,并在癌症免疫治疗中展现出巨大的潜力。NK 细胞的传统来源,如外周血,受到可用性和供体变异性的限制。此外,体外扩增可能导致功能衰竭和基因编辑挑战。本研究旨在利用诱导多能干细胞 (iPSC) 技术提供一致且可扩展的 NK 细胞来源,克服传统来源的限制,并增强癌症免疫治疗应用的潜力。我们使用重编程技术开发了人胎盘来源的 iPSC 系。随后,引入了一种优化的两步分化方案来产生高纯度的 NK 细胞。首先,iPSC 通过无旋转胚胎体 (EB) 分化为造血样干细胞。随后,将 EBs 转移到超低附着平板上以诱导 NK 细胞分化。iPSC 衍生的 NK (iNK) 细胞在 RNA 和蛋白质水平上表达常见的 NK 细胞标志物 (NKp46、NKp30、NKp44、CD16 和 eomesodermin)。iNK 细胞对冷冻保存具有显著的抗性,并表现出增强的细胞毒性。嵌合抗原受体 (CAR) 构建体的掺入进一步增强了它们的细胞毒性潜力。本研究例证了以高纯度和增强的功能能力生成 iNK 细胞的可行性,以及它们对冷冻保存的增强抗性和通过 CAR 表达增强细胞毒性的潜力。我们的研究结果为开发潜在的细胞免疫疗法提供了有前途的途径,突出了 iPSC 技术在克服与传统 NK 细胞来源相关的挑战方面的关键作用。

相似文献

[1]
A scalable, spin-free approach to generate enhanced induced pluripotent stem cell-derived natural killer cells for cancer immunotherapy.

Immunol Cell Biol. 2024-11

[2]
Phenotypic and transcriptomic profiling of induced pluripotent stem cell (iPSC)-derived NK cells and their cytotoxicity against cancers.

Stem Cell Res Ther. 2024-11-13

[3]
Non-clinical efficacy, safety and stable clinical cell processing of induced pluripotent stem cell-derived anti-glypican-3 chimeric antigen receptor-expressing natural killer/innate lymphoid cells.

Cancer Sci. 2020-3-31

[4]
Differentiation of natural killer cells from induced pluripotent stem cells under defined, serum- and feeder-free conditions.

Cytotherapy. 2021-10

[5]
Robust differentiation of NK cells from MSLN.CAR-IL-15-engineered human iPSCs with enhanced antitumor efficacy against solid tumors.

Sci Adv. 2025-5-2

[6]
iPSC-Derived Natural Killer Cell Therapies - Expansion and Targeting.

Front Immunol. 2022

[7]
An Improved Method to Produce Clinical-Scale Natural Killer Cells from Human Pluripotent Stem Cells.

Methods Mol Biol. 2019

[8]
Off-the-shelf cell therapy with induced pluripotent stem cell-derived natural killer cells.

Semin Immunopathol. 2018-10-25

[9]
Targeted integration of EpCAM-specific CAR in human induced pluripotent stem cells and their differentiation into NK cells.

Stem Cell Res Ther. 2021-11-21

[10]
Natural Killer Cells Generated From Human Induced Pluripotent Stem Cells Mature to CD56CD16NKp80 and Express KIR2DL2/DL3 and KIR3DL1.

Front Immunol. 2021

引用本文的文献

[1]
Engineered iPSC-derived natural killer cells: recent innovations in translational innate anti-cancer immunotherapy.

Clin Transl Immunology. 2025-7-10

[2]
Characterization of ex vivo expanded natural killer cells for cancer immunotherapy.

Immunol Cell Biol. 2025-8

[3]
Current Landscape and Future Directions in Cancer Immunotherapy: Therapies, Trials, and Challenges.

Cancers (Basel). 2025-2-27

[4]
Enhancement of anti-sarcoma immunity by NK cells engineered with mRNA for expression of a EphA2-targeted CAR.

Clin Transl Med. 2025-1

[5]
The current socioeconomic and regulatory landscape of immune effector cell therapies.

Front Med (Lausanne). 2024-12-4

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