Genes limiting T cell antitumor activity may serve as therapeutic targets. It has not been systematically studied whether there are regulators that uniquely or broadly contribute to T cell fitness. We perform genome-scale CRISPR-Cas9 knockout screens in primary CD8 T cells to uncover genes negatively impacting fitness upon three modes of stimulation: (1) intense, triggering activation-induced cell death (AICD); (2) acute, triggering expansion; (3) chronic, causing dysfunction. Besides established regulators, we uncover genes controlling T cell fitness either specifically or commonly upon differential stimulation. Dap5 ablation, ranking highly in all three screens, increases translation while enhancing tumor killing. Loss of Icam1-mediated homotypic T cell clustering amplifies cell expansion and effector functions after both acute and intense stimulation. Lastly, Ctbp1 inactivation induces functional T cell persistence exclusively upon chronic stimulation. Our results functionally annotate fitness regulators based on their unique or shared contribution to traits limiting T cell antitumor activity.

限制T细胞抗肿瘤活性的基因可以作为治疗靶点。目前尚未系统研究是否存在对 T 细胞适应性有独特或广泛贡献的调节因子。我们在原代 CD8 T 细胞中进行基因组规模的 CRISPR-Cas9 敲除筛选，以发现对三种刺激模式产生负面影响的基因：(1) 强烈的、触发激活诱导的细胞死亡 (AICD)； (2) 急性、引发扩张； (3)慢性的，引起功能障碍。除了已建立的调节因子外，我们还发现了在差异刺激下特异性或普遍控制 T 细胞适应性的基因。Dap5消融在所有三个筛选中均排名靠前，可增加翻译，同时增强肿瘤杀伤能力。Icam1介导的同型 T 细胞簇的丧失会在急性和强烈刺激后放大细胞扩增和效应器功能。最后，Ctbp1失活仅在慢性刺激下诱导功能性 T 细胞持续存在。我们的结果根据适应性调节因子对限制 T 细胞抗肿瘤活性的特征的独特或共同贡献，对它们进行功能注释。