miR-142-5p promotes TSCM differentiation and suppresses progressive T-cell maturation via targeting PRKCB.

Introduction: Adoptive cellular immunotherapy (ACT) such as CAR‑T therapy holds promise for cancer treatment. However, genetically engineered T cells often undergo terminal differentiation during ex vivo expansion, which limits their persistence and antitumor efficacy Methods: Bioinformatics analysis suggested that miR‑142‑5p target genes are involved in transcription regulation. We identified four candidate targets through reverse enrichment analysis. Dual‑luciferase reporter assays were used to validate direct targets. Functional studies were then performed in T cells overexpressing miR‑142‑5p to assess expression of differentiation‑associated and effector‑related genes, surface markers (CCR7, CD62L, CD95), cell subset proportions (T Results: Dual‑luciferase reporter assays confirmed PRKCB as a direct target of miR‑142‑5p, and miR‑142‑5p suppressed PRKCB expression in T cells. miR‑142‑5p overexpression upregulated early differentiation‑associated genes (LEF1, CD62L, CCR7) and the anti‑apoptotic gene BCL2, while downregulating late differentiation‑associated genes (KLRG1, EOMES, PDCD1) and effector function‑related genes (GZMB, PRF1). Consistently, it enhanced early differentiation markers (CCR7, CD62L) and reduced the late marker CD95. It also increased T Discussion: In summary, miR‑142‑5p inhibits progressive T‑cell differentiation by directly targeting PRKCB, helping maintain an early‑differentiated phenotype. This offers a potential strategy to improve the persistence and efficacy of adoptive T‑cell‑based immunotherapies.