Cellular programming of naïve T cells can improve the efficacy of adoptive T-cell therapy. However, the current ex vivo engineering of T cells requires the pre-activation of T cells, which causes them to lose their naïve state. In this study, cationic-polymer-functionalized nanowires were used to pre-program the fate of primary naïve CD8⁺ T cells to achieve a therapeutic response in vivo. This was done by delivering single or multiple microRNAs to primary naïve mouse and human CD8⁺ T cells without pre-activation. The use of nanowires further allowed for the delivery of large, whole lentiviral particles with potential for long-term integration. The combination of deletion and overexpression of miR-29 and miR-130 impacted the ex vivo T-cell differentiation fate from the naïve state. The programming of CD8⁺ T cells using nanowire-delivered co-delivery of microRNAs resulted in the modulation of T-cell fitness by altering the T-cell proliferation, phenotypic and transcriptional regulation, and secretion of effector molecules. Moreover, the in vivo adoptive transfer of murine CD8⁺ T cells programmed through the nanowire-mediated dual delivery of microRNAs provided enhanced immune protection against different types of intracellular pathogen (influenza and Listeria monocytogenes). In vivo analyses demonstrated that the simultaneous alteration of miR-29 and miR-130 levels in naïve CD8⁺ T cells reduces the persistence of canonical memory T cells whereas increases the population of short-lived effector T cells. Nanowires could potentially be used to modulate CD8⁺ T-cell differentiation and achieve a therapeutic response in vivo without the need for pre-activation.

初始 T 细胞的细胞编程可以提高过继性 T 细胞疗法的疗效。然而，目前 T 细胞的离体工程需要 T 细胞的预激活，这会导致它们失去原始状态。在这项研究中，阳离子聚合物功能化纳米线用于预编程原代幼稚 CD8 ⁺ T 细胞的命运，以实现体内治疗反应。这是通过将单个或多个 microRNA 传递至原代幼稚小鼠和人类 CD8 ⁺ T 细胞而无需预激活来完成的。纳米线的使用进一步允许递送具有长期整合潜力的大的、完整的慢病毒颗粒。 miR-29 和 miR-130 的缺失和过度表达的组合影响了离体 T 细胞从初始状态的分化命运。使用纳米线共同传递的 microRNA 对 CD8 ⁺ T 细胞进行编程，通过改变 T 细胞增殖、表型和转录调节以及效应分子的分泌来调节 T 细胞的适应性。此外，通过纳米线介导的微小RNA双重递送编程的小鼠CD8 ⁺ T细胞的体内过继转移提供了针对不同类型的细胞内病原体（流感和单核细胞增生李斯特菌）的增强的免疫保护。体内分析表明，初始 CD8 ⁺ T 细胞中 miR-29 和 miR-130 水平的同时改变会降低典型记忆 T 细胞的持久性，同时增加短寿命效应 T 细胞的数量。纳米线有可能用于调节 CD8 ⁺ T 细胞分化并在体内实现治疗反应，而无需预激活。