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In vivo screens using a selective CRISPR antigen removal lentiviral vector system reveal immune dependencies in renal cell carcinoma
Immunity ( IF 32.4 ) Pub Date : 2021-01-25 , DOI: 10.1016/j.immuni.2021.01.001
Juan Dubrot 1 , Sarah Kate Lane-Reticker 1 , Emily A Kessler 1 , Austin Ayer 1 , Gargi Mishra 1 , Clara H Wolfe 1 , Margaret D Zimmer 1 , Peter P Du 1 , Animesh Mahapatra 1 , Kyle M Ockerman 1 , Thomas G R Davis 1 , Ian C Kohnle 1 , Hans W Pope 1 , Peter M Allen 1 , Kira E Olander 1 , Arvin Iracheta-Vellve 1 , John G Doench 1 , W Nicholas Haining 2 , Kathleen B Yates 3 , Robert T Manguso 3
Affiliation  

CRISPR-Cas9 genome engineering has increased the pace of discovery for immunology and cancer biology, revealing potential therapeutic targets and providing insight into mechanisms underlying resistance to immunotherapy. However, endogenous immune recognition of Cas9 has limited the applicability of CRISPR technologies in vivo. Here, we characterized immune responses against Cas9 and other expressed CRISPR vector components that cause antigen-specific tumor rejection in several mouse cancer models. To avoid unwanted immune recognition, we designed a lentiviral vector system that allowed selective CRISPR antigen removal (SCAR) from tumor cells. The SCAR system reversed immune-mediated rejection of CRISPR-modified tumor cells in vivo and enabled high-throughput genetic screens in previously intractable models. A pooled in vivo screen using SCAR in a CRISPR-antigen-sensitive renal cell carcinoma revealed resistance pathways associated with autophagy and major histocompatibility complex class I (MHC class I) expression. Thus, SCAR presents a resource that enables CRISPR-based studies of tumor-immune interactions and prevents unwanted immune recognition of genetically engineered cells, with implications for clinical applications.



中文翻译:

使用选择性 CRISPR 抗原去除慢病毒载体系统进行体内筛选揭示肾细胞癌的免疫依赖性

CRISPR-Cas9 基因组工程加快了免疫学和癌症生物学的发现步伐,揭示了潜在的治疗靶点,并提供了对免疫疗法耐药性机制的深入了解。然而,Cas9 的内源性免疫识别限制了 CRISPR 技术在体内的适用性。在这里,我们表征了针对 Cas9 和其他表达的 CRISPR 载体成分的免疫反应,这些成分在几种小鼠癌症模型中引起抗原特异性肿瘤排斥。为了避免不需要的免疫识别,我们设计了一种慢病毒载体系统,允许从肿瘤细胞中选择性去除 CRISPR 抗原 (SCAR)。SCAR系统在体内逆转免疫介导的CRISPR修饰肿瘤细胞排斥反应并在以前难以处理的模型中启用高通量遗传筛选。甲汇集在体内使用SCAR在CRISPR抗原敏感的肾细胞癌揭示了与自噬和主要组织相容性复合体I类(MHC I类)表达相关的电阻通路屏。因此,SCAR 提供了一种资源,可以实现基于 CRISPR 的肿瘤-免疫相互作用研究,并防止对基因工程细胞进行不必要的免疫识别,对临床应用具有影响。

更新日期:2021-03-09
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