Fritz Ashley L, Mao Sunnie R, West Mary G, Schaffer David V
Department of Chemical and Biomolecular Engineering, University of California, Berkeley, California, 94720.
Biotechnol Bioeng. 2015 Jan;112(1):209-19. doi: 10.1002/bit.25336. Epub 2014 Sep 2.
The induction of pluripotency from adult cells has enormous potential in regenerative medicine. While initial efforts to study mechanisms and improve efficiency of induced pluripotent stem cell (iPSC) reprogramming focused on the direct roles of transcriptional regulators, increasing evidence indicates that cellular signal transduction pathways can modulate this process. Here, we present a medium-throughput system to study the effect of signaling pathways on the early stages of reprogramming. We generated a set of lentiviral vectors encoding 38 genes that upregulate or downregulate major signal transduction pathways and quantified each signaling factor's effect on reprogramming. This approach confirmed the role of several factors previously implicated in reprogramming, as well as identified several GTPases-factors that to date have not been largely studied in reprogramming-that improve or hinder iPSC reprogramming. In addition, this methodology is useful in determining new targets for enhancing pluripotency reprogramming, lineage reprogramming, and/or cell differentiation.
从成体细胞诱导多能性在再生医学中具有巨大潜力。虽然最初研究诱导多能干细胞（iPSC）重编程机制和提高其效率的努力主要集中在转录调节因子的直接作用上，但越来越多的证据表明细胞信号转导通路可以调节这一过程。在此，我们展示了一种中通量系统，用于研究信号通路对重编程早期阶段的影响。我们构建了一组编码38个基因的慢病毒载体，这些基因可上调或下调主要信号转导通路，并量化了每个信号因子对重编程的影响。这种方法证实了先前与重编程相关的几个因子的作用，同时还鉴定出了几个GTP酶因子（这些因子迄今为止在重编程研究中尚未得到广泛研究），它们可促进或阻碍iPSC重编程。此外，这种方法有助于确定增强多能性重编程、谱系重编程和/或细胞分化的新靶点。
