美国加州大学旧金山分校Julia Carnevale小组取得一项新突破。他们研制了实体肿瘤中人类T细胞的体内全基因组CRISPR筛选。相关论文于2026年8月12日发表在《自然》杂志上。

在这里，该课题组开发了一种体内模型，可以有效地从实体肿瘤中恢复人类T细胞，允许使用少量小鼠进行全基因组CRISPR筛选。与脾T细胞相比，该模型的肿瘤浸润性T细胞表现出功能障碍的特征，创造了理想的筛选环境。课题组人员进行了两个全基因组CRISPR敲除筛选，以鉴定细胞内T细胞丰度和效应功能的调节因子。丰度筛选显示P2RY8–Gα13 GPCR信号轴是T细胞肿瘤浸润的负调控因子。效应功能筛选发现GNAS是肿瘤中T细胞功能障碍的关键驱动因素，其产物Gαs作为多个GPCR下游的聚合节点，感知不同的抑制配体。

在嵌合抗原受体（CAR）和T细胞受体（TCR）系统中，敲除GNAS使T细胞对多种抑制信号产生抗性，并显著提高了不同实体肿瘤模型的疗效。P2RY8-GNAS的组合敲除进一步增强了对肿瘤的控制，表明互补的体内筛选可以识别出正交靶点，其联合编辑可以提高治疗效力。这种灵活的、可扩展的平台可以用于系统地发现遗传策略，以改善实体肿瘤T细胞疗法。

研究人员表示，在人类T细胞中进行大规模CRISPR筛选，对于识别增强细胞免疫治疗的遗传修饰具有重要的前景。然而，在实体肿瘤中，许多T细胞表现的调节因子并没有在体外被发现。在荷瘤小鼠体内进行筛选更具有生理性，但由于瘤内T细胞恢复较低而受到限制。

附：英文原文

Title: In vivo genome-wide CRISPR screens of human T cells in solid tumours

Author: Liu, Qi, Chen, Peixin Amy, Urs, Esha, Zhang, Shimin, Arce, Maya M., Wang, Charlotte H., Yan, Jun, Nguyen, Vinh Q., Li, Zhongmei, Seo, Jin, Kale, Nupura, Peng, Fanglue, Luo, Yikai, Goudy, Laine, LaFlam, Taylor N., Zhong, Haixia, Modak, Chandrima, Dann, Emma, Jung, Jae Hyung, Kirane, Amanda, Warner, Allison Betof, Quach, Boi Bryant, Good, Zinaida, Shy, Brian R., Shifrut, Eric, Bapat, Sagar P., Allen, Greg M., Eyquem, Justin, Fuh, Katherine, Dodgson, Stacie E., Cyster, Jason G., Marson, Alexander, Carnevale, Julia

Issue&Volume: 2026-08-12

Abstract: Large-scale CRISPR screening in human T cells holds significant promise for identifying genetic modifications that enhance cellular immunotherapy. Yet, many regulators of T cell performance in solid tumours are not revealed in vitro1,2. In vivo screening in tumour-bearing mice is more physiological but has been limited by low intratumoural T cell recovery. Here we developed an in vivo model that efficiently recovers human T cells from solid tumours, permitting genome-wide CRISPR screens with few mice. Tumour-infiltrating T cells from this model exhibit hallmarks of dysfunction compared with splenic T cells, creating an ideal screening context. We performed two genome-wide CRISPR knockout screens to identify regulators of intratumoural T cell abundance and effector function. The abundance screen revealed the P2RY8–Gα13 GPCR signalling axis as a negative regulator of T cell tumour infiltration. The effector function screen identified GNAS as a key driver of T cell dysfunction in tumours, whose product, Gαs, acts as a convergent node downstream of multiple GPCRs sensing distinct suppressive ligands. Knockout of GNAS rendered T cells resistant to multiple suppressive cues and significantly improved efficacy across diverse solid tumour models in chimeric antigen receptor (CAR) and T cell receptor (TCR) systems. Combinatorial knockout of P2RY8–GNAS further enhanced tumour control, demonstrating that complementary in vivo screens can identify orthogonal targets whose combined editing improves therapeutic potency. This flexible, scalable platform can be adapted for systematic discovery of genetic strategies to improve solid tumour T cell therapies.

DOI: 10.1038/s41586-026-10906-9

Source: https://www.nature.com/articles/s41586-026-10906-9