Generation of Pulmonary Endothelial Progenitor Cells for Cell-based Therapy Using Interspecies Mouse-Rat Chimeras.
Generation of Pulmonary Endothelial Progenitor Cells for Cell-based Therapy Using Interspecies Mouse-Rat Chimeras.
复制标题
使用种间小鼠-大鼠嵌合体生成肺内皮祖细胞用于基于细胞的治疗。
DOI：
10.1164/rccm.202003-0758oc
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发表时间：
2021
影响因子：
24.7
通讯作者：
Kalinichenko,VladimirV
中科院分区：
文献类型：
--
作者：
Wang,Guolun;Wen,Bingqiang;Ren,Xiaomeng;Li,Enhong;Zhang,Yufang;Guo,Minzhe;Xu,Yan;Whitsett,JeffreyA;Kalin,TanyaV;Kalinichenko,VladimirV
理由：虽然肺内皮祖细胞（EPC）有望用于新生儿肺部疾病的细胞疗法，但EPC是否可以源自多能胚胎干细胞（ESC）或诱导多能干细胞仍不清楚。目的：研究肺内皮祖细胞的异质性并从多能ESC中衍生功能性EPC。方法：利用新生儿和小鼠肺的单细胞RNA测序来鉴定肺内皮祖细胞的异质性。肺 EPC 的异质性。使用 CRISPR/Cas9 基因编辑对小鼠肺部 EPC 进行基因标记和纯化。将细胞移植到具有 S52F Foxf1 突变的新生小鼠后，对 EPC 的功能特性进行了评估，这是一种肺泡毛细血管发育不良伴肺静脉错位 (ACDMPV) 的小鼠模型。通过囊胚互补产生种间小鼠-大鼠嵌合体，从多能 ESC 产生 EPC，用于 ACDMPV 小鼠的细胞治疗。测量和主要结果：我们鉴定了一个独特的 EPC 群体，FOXF1+cKIT+EPC，作为最近描述的表达 SMAD7、ZBTB20、NFIA 和 DLL4 但缺乏成熟动脉、静脉和淋巴标记物。 FOXF1+cKIT+gCAP 在 ACDMPV 中减少，并且它们的转录组特征在小鼠和人类肺部中是保守的。将细胞移植到 ACDMPV 小鼠的新生循环中后，FOXF1+cKIT+gCAP 植入肺血管系统中，刺激血管生成，改善氧合，并防止肺泡简化。 FOXF1+cKIT+gCAP 是由种间嵌合体中的 ESC 产生的，完全有能力刺激 ACDMPV 小鼠的新生肺血管生成和肺泡化。结论：使用供体或 ESC/诱导的多能干细胞衍生的 FOXF1+cKIT+内皮祖细胞的细胞治疗可考虑用于治疗人类 ACDMPV。
Rationale: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.Objectives:To investigate the heterogeneity of pulmonary EPCs and derive functional EPCs from pluripotent ESCs.Methods: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 withS52F Foxf1mutation, 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.Measurements and Main Results:We identified a unique population of EPCs, FOXF1+cKIT+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. FOXF1+cKIT+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, FOXF1+cKIT+gCAPs engraft into the pulmonary vasculature, stimulate angiogenesis, improve oxygenation, and prevent alveolar simplification. FOXF1+cKIT+gCAPs, produced from ESCs in interspecies chimeras, are fully competent to stimulate neonatal lung angiogenesis and alveolarization in ACDMPV mice.Conclusions:Cell-based therapy using donor or ESC/induced pluripotent stem cell–derived FOXF1+cKIT+endothelial progenitors may be considered for treatment of human ACDMPV.