Department of Microbiology and Immunology, David H. Smith Center for Vaccine Biology and Immunology, University of Rochester, Rochester, NY, USA.
Department of Pathology, University of Rochester Medical Center, Rochester, NY, USA.
Nat Immunol. 2023 Jun;24(6):1007-1019. doi: 10.1038/s41590-023-01498-x. Epub 2023 Apr 17.
Adoptive transfer of genetically engineered chimeric antigen receptor (CAR) T cells is becoming a promising treatment option for hematological malignancies. However, T cell immunotherapies have mostly failed in individuals with solid tumors. Here, with a CRISPR-Cas9 pooled library, we performed an in vivo targeted loss-of-function screen and identified ST3 β-galactoside α-2,3-sialyltransferase 1 (ST3GAL1) as a negative regulator of the cancer-specific migration of CAR T cells. Analysis of glycosylated proteins revealed that CD18 is a major effector of ST3GAL1 in activated CD8 T cells. ST3GAL1-mediated glycosylation induces the spontaneous nonspecific tissue sequestration of T cells by altering lymphocyte function-associated antigen-1 (LFA-1) endocytic recycling. Engineered CAR T cells with enhanced expression of βII-spectrin, a central LFA-1-associated cytoskeleton molecule, reversed ST3GAL1-mediated nonspecific T cell migration and reduced tumor growth in mice by improving tumor-specific homing of CAR T cells. These findings identify the ST3GAL1-βII-spectrin axis as a major cell-intrinsic program for cancer-targeting CAR T cell migration and as a promising strategy for effective T cell immunotherapy.
过继输注基因工程嵌合抗原受体（CAR）T 细胞正在成为血液恶性肿瘤的一种很有前途的治疗选择。然而，T 细胞免疫疗法在实体瘤患者中大多失败。在这里，我们使用 CRISPR-Cas9 文库进行了体内靶向功能丧失筛选，并鉴定 ST3 β-半乳糖苷 α-2,3-唾液酸转移酶 1（ST3GAL1）为 CAR T 细胞对癌症特异性迁移的负调节因子。糖基化蛋白分析表明，CD18 是激活的 CD8 T 细胞中 ST3GAL1 的主要效应因子。ST3GAL1 介导的糖基化通过改变淋巴细胞功能相关抗原-1（LFA-1）内吞循环，诱导 T 细胞自发的非特异性组织隔离。通过增强 CAR T 细胞中βII- spectrin 的表达，一种中央 LFA-1 相关细胞骨架分子，可逆转 ST3GAL1 介导的非特异性 T 细胞迁移，并通过改善 CAR T 细胞对肿瘤的特异性归巢来减少小鼠肿瘤生长。这些发现确定了 ST3GAL1-βII- spectrin 轴作为癌症靶向 CAR T 细胞迁移的主要细胞内在程序，并为有效的 T 细胞免疫治疗提供了有前途的策略。
