多谱系筛选揭示 mTORC1 抑制增强人类多能干细胞中胚层和造血祖细胞的生成。
A Multi-Lineage Screen Reveals mTORC1 Inhibition Enhances Human Pluripotent Stem Cell Mesendoderm and Blood Progenitor Production.
机构信息
Institute of Biomaterials and Biomedical Engineering, University of Toronto, Toronto, ON M5S 3G9, Canada.
Institute of Biomaterials and Biomedical Engineering, University of Toronto, Toronto, ON M5S 3G9, Canada; Department of Chemical Engineering and Applied Chemistry, University of Toronto, Toronto, ON M5S 3E5, Canada.
出版信息
Stem Cell Reports. 2016 May 10;6(5):679-691. doi: 10.1016/j.stemcr.2016.04.003. Epub 2016 Apr 28.
Human pluripotent stem cells (hPSCs) exist in heterogeneous micro-environments with multiple subpopulations, convoluting fate-regulation analysis. We patterned hPSCs into engineered micro-environments and screened responses to 400 small-molecule kinase inhibitors, measuring yield and purity outputs of undifferentiated, neuroectoderm, mesendoderm, and extra-embryonic populations. Enrichment analysis revealed mammalian target of rapamycin (mTOR) inhibition as a strong inducer of mesendoderm. Dose responses of mTOR inhibitors such as rapamycin synergized with Bone Morphogenetic protein 4 (BMP4) and activin A to enhance the yield and purity of BRACHYURY-expressing cells. Mechanistically, small interfering RNA knockdown of RAPTOR, a component of mTOR complex 1, phenocopied the mesendoderm-enhancing effects of rapamycin. Functional analysis during mesoderm and endoderm differentiation revealed that mTOR inhibition increased the output of hemogenic endothelial cells 3-fold, with a concomitant enhancement of blood colony-forming cells. These data demonstrate the power of our multi-lineage screening approach and identify mTOR signaling as a node in hPSC differentiation to mesendoderm and its derivatives.
人类多能干细胞（hPSCs）存在于具有多种亚群的异质微环境中，这使得对其命运调控的分析变得复杂。我们将 hPSCs 设计成工程化的微环境，并筛选对 400 种小分子激酶抑制剂的反应，测量未分化、神经外胚层、中胚层和胚胎外群体的产量和纯度输出。富集分析显示雷帕霉素靶蛋白（mTOR）抑制是中胚层的强烈诱导剂。mTOR 抑制剂（如雷帕霉素）的剂量反应与骨形态发生蛋白 4（BMP4）和激活素 A 协同作用，可提高 BRACHYURY 表达细胞的产量和纯度。在机制上，mTOR 复合物 1 的组成部分 RAPTOR 的小干扰 RNA 敲低模拟了雷帕霉素对中胚层增强的作用。中胚层和内胚层分化过程中的功能分析表明，mTOR 抑制将造血内皮细胞的产量提高了 3 倍，同时增加了血液集落形成细胞的数量。这些数据证明了我们多谱系筛选方法的强大功能，并确定 mTOR 信号作为 hPSC 向中胚层及其衍生物分化的节点。
Zotero 插件