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对肿瘤浸润原代NK细胞进行体内AAV-SB-CRISPR筛选确定CAR-NK疗法的基因检查点
作者:小柯机器人 发布时间:2024/6/29 16:38:20

美国耶鲁大学陈斯迪等研究人员合作发现,对肿瘤浸润原代自然杀伤(NK)细胞进行体内AAV-SB-CRISPR筛选确定CAR-NK疗法的基因检查点。相关论文于2024年6月25日在线发表在《自然—生物技术》杂志上。

研究人员利用体内腺相关病毒(AAV)-SB(睡美人)-CRISPR(成簇间隔短回文重复序列)筛选技术,在四种实体瘤小鼠模型中对肿瘤浸润NK(TINK)细胞进行了无偏倚的功能图谱绘制。
 
与此同时,研究人员对TINK细胞的单细胞转录组图谱进行了鉴定,发现了以前未曾探索过的NK细胞亚群和不同表达的TINK基因。在小鼠原代NK细胞和人类嵌合抗原受体(CAR)-NK细胞中,CALHM2-基因敲除(KO)的NK细胞显示出更强的细胞毒性和肿瘤浸润能力。CALHM2 mRNA逆转了CALHM2-KO表型。人类原代NK细胞中的CALHM2 KO能增强其细胞毒性、脱颗粒性和细胞因子的产生。
 
转录组学分析显示,CALHM2-KO改变了基线和刺激条件下的基因和通路。在对未修饰的CAR-NK细胞有抗药性的实体瘤模型中,CALHM2-KO CAR-NK细胞显示出了强大的体内抗肿瘤功效。这些数据确定了自然限制NK细胞功能的内源性基因检查点,并证明了CALHM2 KO可用于设计基于NK细胞的增强型免疫疗法。

研究人员表示,NK细胞具有抗癌的临床潜力;然而,多种限制因素阻碍了NK细胞疗法的成功。

附:英文原文

Title: In vivo AAV–SB-CRISPR screens of tumor-infiltrating primary NK cells identify genetic checkpoints of CAR-NK therapy

Author: Peng, Lei, Renauer, Paul A., Sferruzza, Giacomo, Yang, Luojia, Zou, Yongji, Fang, Zhenghao, Park, Jonathan J., Chow, Ryan D., Zhang, Yueqi, Lin, Qianqian, Bai, Meizhu, Sanchez, Angelica, Zhang, Yongzhan, Lam, Stanley Z., Ye, Lupeng, Chen, Sidi

Issue&Volume: 2024-06-25

Abstract: Natural killer (NK) cells have clinical potential against cancer; however, multiple limitations hinder the success of NK cell therapy. Here, we performed unbiased functional mapping of tumor-infiltrating NK (TINK) cells using in vivo adeno-associated virus (AAV)–SB (Sleeping Beauty)-CRISPR (clustered regularly interspaced short palindromic repeats) screens in four solid tumor mouse models. In parallel, we characterized single-cell transcriptomic landscapes of TINK cells, which identified previously unexplored subpopulations of NK cells and differentially expressed TINK genes. As a convergent hit, CALHM2-knockout (KO) NK cells showed enhanced cytotoxicity and tumor infiltration in mouse primary NK cells and human chimeric antigen receptor (CAR)-NK cells. CALHM2 mRNA reversed the CALHM2-KO phenotype. CALHM2 KO in human primary NK cells enhanced their cytotoxicity, degranulation and cytokine production. Transcriptomics profiling revealed CALHM2-KO-altered genes and pathways in both baseline and stimulated conditions. In a solid tumor model resistant to unmodified CAR-NK cells, CALHM2-KO CAR-NK cells showed potent in vivo antitumor efficacy. These data identify endogenous genetic checkpoints that naturally limit NK cell function and demonstrate the use of CALHM2 KO for engineering enhanced NK cell-based immunotherapies.

DOI: 10.1038/s41587-024-02282-4

Source: https://www.nature.com/articles/s41587-024-02282-4

期刊信息

Nature Biotechnology:《自然—生物技术》,创刊于1996年。隶属于施普林格·自然出版集团,最新IF:68.164
官方网址:https://www.nature.com/nbt/
投稿链接:https://mts-nbt.nature.com/cgi-bin/main.plex