Center for Lung Regenerative Medicine.
Division of Pulmonary Biology, and.
Am J Respir Crit Care Med. 2021 Aug 1;204(3):326-338. doi: 10.1164/rccm.202003-0758OC.
Although pulmonary endothelial progenitor cells (EPCs) hold promise for cell-based therapies for neonatal pulmonary disorders, whether EPCs can be derived from pluripotent embryonic stem cells (ESCs) or induced pluripotent stem cells remains unknown. To investigate the heterogeneity of pulmonary EPCs and derive functional EPCs from pluripotent ESCs. Single-cell RNA sequencing of neonatal human and mouse lung was used to identify the heterogeneity of pulmonary EPCs. CRISPR/Cas9 gene editing was used to genetically label and purify mouse pulmonary EPCs. Functional properties of the EPCs were assessed after cell transplantation into neonatal mice with mutation, a mouse model of alveolar capillary dysplasia with misalignment of pulmonary veins (ACDMPV). Interspecies mouse-rat chimeras were produced through blastocyst complementation to generate EPCs from pluripotent ESCs for cell therapy in ACDMPV mice. We identified a unique population of EPCs, FOXF1cKIT EPCs, as a subset of recently described general capillary cells (gCAPs) expressing SMAD7, ZBTB20, NFIA, and DLL4 but lacking mature arterial, venous, and lymphatic markers. FOXF1cKIT gCAPs are reduced in ACDMPV, and their transcriptomic signature is conserved in mouse and human lungs. After cell transplantation into the neonatal circulation of ACDMPV mice, FOXF1cKIT gCAPs engraft into the pulmonary vasculature, stimulate angiogenesis, improve oxygenation, and prevent alveolar simplification. FOXF1cKIT gCAPs, produced from ESCs in interspecies chimeras, are fully competent to stimulate neonatal lung angiogenesis and alveolarization in ACDMPV mice. Cell-based therapy using donor or ESC/induced pluripotent stem cell-derived FOXF1cKIT endothelial progenitors may be considered for treatment of human ACDMPV.
尽管肺内皮祖细胞（EPC）有望成为治疗新生儿肺部疾病的细胞治疗方法，但 EPC 是否可以从多能胚胎干细胞（ESC）或诱导多能干细胞（iPSC）中获得仍然未知。为了研究肺 EPC 的异质性并从多能 ESC 中获得功能性 EPC。使用单细胞 RNA 测序对新生人和鼠肺进行了研究，以鉴定肺 EPC 的异质性。使用 CRISPR/Cas9 基因编辑对小鼠肺 EPC 进行遗传标记和纯化。在 突变的新生小鼠中进行细胞移植后，评估 EPC 的功能特性，该突变是肺泡毛细血管发育不良伴肺静脉错位（ACDMPV）的小鼠模型。通过胚泡互补产生种间鼠-大鼠嵌合体，以从多能 ESC 中产生 EPC 用于 ACDMPV 小鼠的细胞治疗。我们鉴定了一种独特的 EPC 群体，FOXF1cKIT EPC，作为最近描述的一般毛细血管细胞（gCAP）的一个子集，表达 SMAD7、ZBTB20、NFIA 和 DLL4，但缺乏成熟的动脉、静脉和淋巴管标志物。FOXF1cKIT gCAP 在 ACDMPV 中减少，其转录组特征在小鼠和人类肺部中是保守的。在 ACDMPV 小鼠的新生循环中进行细胞移植后，FOXF1cKIT gCAP 植入肺血管，刺激血管生成，改善氧合，并预防肺泡简化。从种间嵌合体中的 ESC 产生的 FOXF1cKIT gCAP 完全有能力刺激 ACDMPV 小鼠的新生肺血管生成和肺泡化。使用供体或 ESC/iPSC 衍生的 FOXF1cKIT 内皮祖细胞的细胞治疗可考虑用于治疗人类 ACDMPV。
