Ishizuka Shuta, Lai Chen-Yi, Otsu Makoto, Nakauchi Hiromitsu, Nagamune Teruyuki, Kawahara Masahiro
Department of Chemistry and Biotechnology, Graduate School of Engineering , The University of Tokyo , 7-3-1 Hongo , Bunkyo-ku , Tokyo 113-8656 , Japan.
Division of Stem Cell Processing/Stem Cell Bank, Center for Stem Cell Biology and Regenerative Medicine , Institute of Medical Science, The University of Tokyo , 4-6-1, Shirokanedai , Minato-ku , Tokyo 108-8639 , Japan.
ACS Synth Biol. 2018 Jul 20;7(7):1709-1714. doi: 10.1021/acssynbio.8b00163. Epub 2018 Jun 21.
The understanding of signaling events is critical for attaining long-term expansion of hematopoietic stem cells ex vivo. In this study, we aim to analyze the contribution of multiple signaling molecules in proliferation of hematopoietic stem cells. To this end, we design a bottom-up engineered receptor with multiple tyrosine motifs, which can recruit multiple signaling molecules of interest. This is followed by a top-down approach, where one of the multiple tyrosine motifs in the bottom-up engineered receptor is functionally knocked out by tyrosine-to-phenylalanine mutation. The combination of these two approaches demonstrates the importance of Shc in cooperation with STAT3 or STAT5 in the proliferation of hematopoietic stem cells. The platform developed herein may be applied for analyzing other cells and/or other cell fate regulation systems.
了解信号传导事件对于在体外实现造血干细胞的长期扩增至关重要。在本研究中，我们旨在分析多种信号分子对造血干细胞增殖的作用。为此，我们设计了一种具有多个酪氨酸基序的自下而上工程化受体，它可以招募多个感兴趣的信号分子。接下来是自上而下的方法，即通过酪氨酸到苯丙氨酸的突变在自下而上工程化受体中的多个酪氨酸基序之一上进行功能敲除。这两种方法的结合证明了Shc与STAT3或STAT5协同作用在造血干细胞增殖中的重要性。本文开发的平台可用于分析其他细胞和/或其他细胞命运调控系统。
