iPSC-Derived Vascularized Human Lung Organoids and Interaction Between Lung Endothelial Cells and Alveolar Epithelial Cells

摘要 人类诱导多能干细胞（iPSC）可用于产生三维肺类器官 结构.然而，大多数肺类器官研究集中在人iPSC衍生的肺上皮亚型。 迄今为止，他们还没有包括人iPSC衍生的内皮细胞，并且系统地解决了关键的问题。 肺血管内皮细胞和血管灌注本身在肺血管生成和成熟中的作用 模拟人类肺部结构的类器官。肺泡单位由两种主要的细胞类型组成- 上皮细胞（EpiC）（总细胞的40-45%）和内皮细胞（EC）（总细胞的45-50%）。我们的关键 支持性数据支持迄今为止被低估的人肺血管内皮细胞的关键作用 细胞在人肺上皮祖细胞分化和血管化人肺形成中的作用 类器官我们建议使用这种通过整合hiPSC衍生的上皮细胞和 目的1：测试内皮细胞来源的内皮细胞的假设， 肺类器官中的血管分泌信号激活Wnt信号传导并介导肺泡单位的成熟， EpiC的相互上分泌信号调节肺EC命运的推论假说，最近产生的 描述了特定的肺EC群体和肺泡水平的肺微血管模式。目标2将测试 假设血管化和灌注的人肺类器官作为一个与肺相关的 简化模型，用于梳理炎性损伤的难以捉摸的信号传导和分子机制， 肺泡单位的水平和损伤的解决。目的3将检验肺EC信号通过 肺泡II型上皮细胞中ACE 2的上调促进SARS-CoV-2进入并感染肺部。 通过关注肺EC和人肺类器官的血管化， 将肺泡上皮细胞纳入该系统将揭示如何在细胞内表达的基本机制。 血管化肺泡单位在健康中的功能是维持体内平衡，以及EC之间如何存在缺陷的串扰。 和肺泡上皮细胞导致炎性肺病。

ABSTRACT Human induced pluripotent stem cells (iPSCs) can be used to generate 3-dimensional lung organoid structures. However, most lung organoid studies have focused on human iPSC-derived lung epithelial subtypes. They have not to date included human iPSC-derived endothelial cells and systematically addressed the critical role of lung vascular endothelial cells and vascular perfusion itself in the generation and maturation of lung organoids which model human lung structures. The alveolar units consist of two predominant cell types – epithelial cells (EpiC)(40-45% of total cells) and endothelial cells (EC) (45-50% of total cells). Our key Supporting Data support the critical and heretofore underestimated role of human lung vascular endothelial cells in guiding differentiation of human lung epithelial progenitor cells and formation of vascularized human lung organoid. We propose to use this novel platform generated by integration of hiPSC-derived epithelial and endothelial cells to address the following aims: Aim 1 tests the hypothesis that endothelial cell-derived angiocrine signals in lung organoids activate Wnt signaling and mediate the maturation of lung alveolar units and the corollary hypothesis that reciprocal epicrine signaling of EpiC regulates lung EC fate, generation of recently described specific lung EC populations and lung microvessel patterning at the level of alveoli. Aim 2 will test the hypothesis that the vascularized and perfused human lung organoid serves as a translationally relevant reductionist model for teasing apart the elusive signaling and molecular mechanisms of inflammatory injury at the level of the alveolar unit and resolution of injury. Aim 3 will test the hypothesis that lung EC signaling through the upregulation of ACE2 in alveolar Type II epithelial cells promotes SARS-CoV-2 entry and infection of lungs. Together the proposed studies through their focus on lung EC and vascularization of human lung organoid and incorporation of alveolar epithelial cells in this system will uncover fundamental mechanisms of how the vascularized alveolar unit functions in health to maintain homeostasis and how defective cross-talk between EC and alveolar epithelial cells contributes to inflammatory lung disease.