奥地利科学院Christoph Bock课题组近日取得一项新成果。经过不懈努力，他们揭示了CRISPR增强CAR T细胞免疫疗法的系统性发现。该研究于2025年9月24日发表于国际一流学术期刊《自然》杂志上。

在这里，该课题组研究人员提出了CELLFIE，一个CRISPR筛选平台，用于增强CAR-T细胞跨越多个临床目标。该研究组在人类原代CAR-T细胞中进行了全基因组筛选，读取了T细胞生物学的关键方面，包括增殖、靶细胞识别、激活、凋亡和自相残杀以及衰竭。在人类白血病异种移植模型中，利用一种新的体内CROP-seq2方法优先筛选hits，建立了几种增强CAR-T细胞功效的基因敲除。最值得注意的是，小组发现RHOG敲除是一种有效的、意想不到的CAR-T细胞增强剂，无论是单独的还是与FAS敲除一起，这在多种体内模型、CAR设计和样本供体以及患者来源的细胞中都得到了验证。为了证明CELLFIE平台的多功能性，课题组人员还进行了组合CRISPR筛选，以鉴定协同基因对和饱和碱基编辑筛选，以表征RHOG变体。总之，课题组发现、验证和生物学表征了CRISPR增强的CAR-T细胞，它们在广泛主题的基准测试中优于标准CAR-T细胞，为优化基于细胞的免疫疗法建立了基础抵抗。

据介绍，嵌合抗原受体（CAR-T）细胞疗法在治疗血癌方面取得了显著的成功，但CAR-T细胞功能障碍仍然是治疗失败的常见原因。

附：英文原文

Title: Systematic discovery of CRISPR-boosted CAR T cell immunotherapies

Author: Datlinger, Paul, Pankevich, Eugenia V., Arnold, Cosmas D., Pranckevicius, Nicole, Lin, Jenny, Romanovskaia, Daria, Schaefer, Moritz, Piras, Francesco, Orts, Anne-Christine, Nemc, Amelie, Biesaga, Paulina N., Chan, Michelle, Neuwirth, Teresa, Artemov, Artem V., Li, Wentao, Ladsttter, Sabrina, Krausgruber, Thomas, Bock, Christoph

Issue&Volume: 2025-09-24

Abstract: Chimeric antigen receptor (CAR) T cell therapy has shown remarkable success in treating blood cancers, but CAR T cell dysfunction remains a common cause of treatment failure1. Here we present CELLFIE, a CRISPR screening platform for enhancing CAR T cells across multiple clinical objectives. We performed genome-wide screens in human primary CAR T cells, with readouts capturing key aspects of T cell biology, including proliferation, target cell recognition, activation, apoptosis and fratricide, and exhaustion. Screening hits were prioritized using a new in vivo CROP-seq2 method in a xenograft model of human leukaemia, establishing several gene knockouts that boost CAR T cell efficacy. Most notably, we discovered that RHOG knockout is a potent and unexpected CAR T cell enhancer, both individually and together with FAS knockout, which was validated across multiple in vivo models, CAR designs and sample donors, and in patient-derived cells. Demonstrating the versatility of the CELLFIE platform, we also conducted combinatorial CRISPR screens to identify synergistic gene pairs and saturation base-editing screens to characterize RHOG variants. In summary, we discovered, validated and biologically characterized CRISPR-boosted CAR T cells that outperform standard CAR T cells in widely used benchmarks, establishing a foundational resource for optimizing cell-based immunotherapies.

DOI: 10.1038/s41586-025-09507-9

Source: https://www.nature.com/articles/s41586-025-09507-9