体外谷氨酰胺抑制的代谢重编程赋予 CAR-T 细胞低分化表型和持久抗肿瘤活性

The inadequate in vivo persistence of chimeric antigen receptor (CAR)-modified T cells has been shown to lead to poor therapeutic efficacy and disease recurrence. In vivo persistence is associated with the differentiation subsets infused, with less differentiated T N or T CM conferring superior renewal capacity and antitumor immunity compared to T EM or T EFF .

However, ex vivo expanded CAR-T cells exhibit phenotypic heterogeneity with majority of T EM or T EFF subsets and very low populations of T N and T CM . The transition of differentiation subsets is closely correlated with T cell metabolism fitness. Effector T cell differentiation from T N or T CM requires glutamine uptake and metabolism.

Using a CD19-specific CAR, we demonstrated that glutamine inhibition by adding the glutamine antagonist 6-Diazo-5-oxo-l-norleucine (DON) into the culture endows CAR-T cells with enhanced mitochondrial OXPHOS utilizing fatty acids and reduced glycolytic activity, and retains more T N or T CM subsets. DON- pretreated CAR-T cells exhibited stronger cytotoxic lysis in vitro and more robust elimination of tumor burdens in vivo.

This study suggests that glutamine inhibition ex vivo would be a potential approach for modulating metabolism and differentiation state to improve the efficacy of CAR-T cell therapy.