Guo Lin, Lin Jiechun, Ren Qiwen, Sun Hao, Wu Yanhua, Ge Haofei, Wu Xiaolan, Lin Lihui, Liang Lining, Li Changpeng, Liu He, Mai Yuangbang, Chu Shilong, Liu Jiadong, Liu Jing, Chen Jiekai, Pei Duanqing, Zheng Hui
Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangdong-Hong Kong Joint Laboratory for Stem Cell and Regenerative Medicine, GIBH-CUHK Joint Research Laboratory on Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, 510530, China.
Joint School of Life Sciences, Guangzhou Medical University, Guangzhou, 511436, China.
Adv Sci (Weinh). 2025 May 31:e15528. doi: 10.1002/advs.202415528.
To investigate the interplay among OCT4, SOX2, and KLF4, the VP16 activation domain is fused to one or multiple of these factors during reprogramming. Fusion of VP16 with OCT4 and SOX2 (OvSvK) significantly increases the efficiency of induced pluripotent stem cell (iPSC) generation compared to other combinations. The enhanced activities of OCT4 and SOX2 directly facilitated reprogramming by activating downstream targets, some of which are involved in cell cycle regulation. This leads to a shortened G1 phase and a shift in cell cycle dynamics toward an iPSCs-like state. Further analysis reveals that these cell cycle alterations reduced H3K27me3 levels on specific genes, thereby promoting reprogramming. Consistently, knockdown of Ccnd1 and Cdkn2a (siCcnd1, siCdkn2a) as well as overexpression of Ccne1 effectively shortened the G1 phase and enhanced reprogramming efficiency. These findings highlight the role of cell cycle modulation in epigenetic remodeling and provide mechanistic insights for optimization during somatic cell reprogramming.
为了研究OCT4、SOX2和KLF4之间的相互作用，在重编程过程中将VP16激活结构域与这些因子中的一个或多个融合。与其他组合相比，VP16与OCT4和SOX2融合（OvSvK）显著提高了诱导多能干细胞（iPSC）的生成效率。OCT4和SOX2增强的活性通过激活下游靶点直接促进重编程，其中一些靶点参与细胞周期调控。这导致G1期缩短，细胞周期动力学向iPSC样状态转变。进一步分析表明，这些细胞周期改变降低了特定基因上的H3K27me3水平，从而促进了重编程。同样，敲低Ccnd1和Cdkn2a（siCcnd1，siCdkn2a）以及过表达Ccne1有效地缩短了G1期并提高了重编程效率。这些发现突出了细胞周期调控在表观遗传重塑中的作用，并为体细胞重编程过程中的优化提供了机制性见解。
