The in vivo proliferation and viability of transfused engineered T cells markedly limits the long-term effect of adoptive cell therapy on tumors. The therapeutic efficacy and proliferative potential of T cells are reported to be dependent on the differentiation status of T cells. The T cells at the early stage of progressive differentiation have a long lifespan, strong proliferative potential, and the ability to reconstruct intact T cell subsets. Thus, they are more suitable for adoptive immunotherapy. Previously, it was difficult to obtain a sufficient number of early differentiated T cells by inhibiting the progressive differentiation of T cells or by two-step programming. A more effective strategy is to directly reprogram and dedifferentiate the easily available terminal effector T (TEFF) cells, which are generated in large numbers, into early T cells. This study attempted to overexpress eight (candidate) early differentiation-specific transcription factors (TFs) (LEF1, KLF7, ID3, EOMES, BCL6, TCF7, FOXP1, and FOXO1) in the TEFF cells, which were activated by in vitro stimulation, to promote dedifferentiation into early T cells. In the mature TEFF cells simultaneously overexpressing these specific TFs, the expression pattern of T cell differentiation markers (CCR7 and CD45RO) exhibited a tendency to change to the pattern observed during early differentiation. The transcriptome analysis revealed that the function of differentially expressed genes was mainly concentrated in the cell cycle, growth and development, and effector function. Moreover, many genes related to early differentiated T cells (such as BCL2 and PIM1) were significantly upregulated, while those related to the effector function of TEFF cells were significantly downregulated (such as GZMB, PRF1, and GNLY). Additionally, the TEFF cells overexpressing characteristic TFs exhibited enhanced anti-apoptotic capabilities and decreased secretion of cytokines (IFN-γ and TNF-α). Based on these results, we believe that the TEFF cells were reprogrammed into a less differentiated state after overexpression of the eight specific TFs.

中文翻译：

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早期T细胞分化特异性转录因子的过表达将终末分化的效应T细胞转化为分化程度较低的状态。

输注改造的T细胞的体内增殖和生存能力明显限制了过继性细胞疗法对肿瘤的长期影响。据报道，T细胞的治疗功效和增殖潜力取决于T细胞的分化状态。进行性分化早期的T细胞具有较长的寿命，强大的增殖潜能以及重建完整T细胞亚群的能力。因此，它们更适合于过继免疫疗法。以前，很难通过抑制T细胞的逐步分化或通过两步编程来获得足够数量的早期分化T细胞。一种更有效的策略是直接对容易产生的大量产生的末端效应子T（TEFF）细胞进行重新编程和去分化，进入早期的T细胞。这项研究试图在体外刺激下激活的TEFF细胞中过表达八个（候选）早期分化特异性转录因子（TF）（LEF1，KLF7，ID3，EOMES，BCL6，TCF7，FOXP1和FOXO1）促进去分化成早期T细胞。在同时过量表达这些特定TF的成熟TEFF细胞中，T细胞分化标志物（CCR7和CD45RO）的表达模式表现出改变为早期分化过程中观察到的模式的趋势。转录组分析表明差异表达基因的功能主要集中在细胞周期，生长发育和效应子功能上。此外，许多与早期分化的T细胞相关的基因（例如BCL2和PIM1）都显着上调，而与TEFF细胞的效应子功能相关的那些则被显着下调（例如GZMB，PRF1和GNLY）。另外，过表达特征性TF的TEFF细胞表现出增强的抗凋亡能力和减少的细胞因子（IFN-γ和TNF-α）的分泌。基于这些结果，我们相信在八个特定TF的过度表达后，TEFF细胞被重新编程为分化程度较低的状态。