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

摘要 人诱导多能干细胞(IPSCs)可用于生成三维肺器官 结构。然而，大多数肺器官研究都集中在人IPSC来源的肺上皮亚型上。 到目前为止，他们还没有包括人IPSC来源的内皮细胞，并系统地解决了关键的 肺血管内皮细胞和血管本身在肺发生和成熟中的作用 模拟人类肺部结构的有机化合物。肺泡单位由两种主要的细胞类型组成- 上皮细胞(EPIC)(占细胞总数的40%~45%)和内皮细胞(EC)(占细胞总数的45%~50%)。我们的钥匙 支持数据支持人肺血管内皮细胞的关键和迄今被低估的作用 细胞在人肺上皮祖细胞分化和血运形成中的作用 有机体。我们建议使用这个新的平台，通过整合HiPSC来源的上皮和 内皮细胞解决以下目标：Aim 1检验内皮细胞起源于 肺组织器官中的血管分泌信号激活Wnt信号，介导肺泡单位的成熟和 EPIC相互表观分泌信号调节肺内皮细胞命运的推论 描述了特定的肺内皮细胞种群和肺泡水平的肺微血管构型。AIM 2将测试 假设带血管的和灌流的人肺器官作为翻译相关 用于梳理炎性损伤难以捉摸的信号和分子机制的简化论模型 肺泡单位的水平和损伤的解决。目标3将测试肺内皮细胞信号传递途径的假设 肺泡II型上皮细胞ACE2表达上调促进SARS-CoV-2侵入和肺部感染。 结合建议的研究，通过他们的重点是肺内皮细胞和人肺器官血管形成和 肺泡上皮细胞在这一系统中的结合将揭示肺泡上皮细胞如何 带血管的肺泡单位功能在健康中维持动态平衡和EC之间的串扰 肺泡上皮细胞参与炎症性肺病的发生。

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.