Origin and Characterization of Alpha Smooth Muscle Actin-Positive Cells During Murine Lung Development
Origin and Characterization of Alpha Smooth Muscle Actin-Positive Cells During Murine Lung Development
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DOI：
10.1002/stem.2615
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发表时间：
2017-06-01
期刊：
影响因子：
5.2
通讯作者：
Bellusci, Saverio
中科院分区：
文献类型：
--
作者：
Moiseenko, Alena;Kheirollahi, Vahid;Bellusci, Saverio
ACTA 2表达鉴定肺气道和血管平滑肌细胞（SMC）以及肺泡肌成纤维细胞（MYF）。表达成纤维细胞生长因子10（Fgf 10）、Wilms肿瘤1（Wt 1）或胶质瘤相关癌基因1（Gli 1）的间充质祖细胞有助于肺发育早期SMC的形成。然而，他们各自的贡献和特异性SMC和/或肺泡MYF谱系仍然存在争议。此外，经历活跃WNT信号传导的间充质细胞的贡献仍然未知。使用Fgf 10（CreERT 2）、Wt 1（CreERT 2）、Gli 1（CreERT 2）和Axin 2（CreERT 2）诱导型驱动细胞系与tdTomato（flox）报告细胞系的组合，定量每个标记祖细胞池沿着SMC和肺泡MYF谱系的各自分化。结果显示，虽然FGF 10（+）和WT 1（+）细胞对SMC谱系的贡献较小，但GLI 1（+）和AXIN 2（+）细胞对SMC和肺泡MYF谱系均有显著贡献，但特异性有限。使用Acta 2-CreERT 2转基因系的谱系追踪显示，在胚胎日（E）11.5标记的ACTA 2（+）细胞在E18.5没有显著扩增以产生新的SMC。然而，在E15.5标记的ACTA 2（+）细胞在E18.5产生肺中的大多数（85%-97%）SMC以及肺实质中的肺泡MYF祖细胞。通过基因阵列技术分离ACTA 2（+）谱系示踪细胞的不同亚群，采用基于细胞分选的方法，在E18.5时鉴定了肺泡MYF祖细胞与气道和血管SMC的转录组学特征。我们的研究结果为进一步的实验建立了一个新的转录景观，解决了ACTA 2（+）细胞不同亚群形成中信号通路的功能。
ACTA2 expression identifies pulmonary airway and vascular smooth muscle cells (SMCs) as well as alveolar myofibroblasts (MYF). Mesenchymal progenitors expressing fibroblast growth factor 10 (Fgf10), Wilms tumor 1 (Wt1), or glioma-associated oncogene 1 (Gli1) contribute to SMC formation from early stages of lung development. However, their respective contribution and specificity to the SMC and/or alveolar MYF lineages remain controversial. In addition, the contribution of mesenchymal cells undergoing active WNT signaling remains unknown. Using Fgf10(CreERT2), Wt1(CreERT2), Gli1(CreERT2), and Axin2(CreERT2) inducible driver lines in combination with a tdTomato(flox) reporter line, the respective differentiation of each pool of labeled progenitor cells along the SMC and alveolar MYF lineages was quantified. The results revealed that while FGF10(+) and WT1(+) cells show a minor contribution to the SMC lineage, GLI1(+) and AXIN2(+) cells significantly contribute to both the SMC and alveolar MYF lineages, but with limited specificity. Lineage tracing using the Acta2-CreERT2 transgenic line showed that ACTA2(+) cells labeled at embryonic day (E)11.5 do not expand significantly to give rise to new SMCs at E18.5. However, ACTA2(+) cells labeled at E15.5 give rise to the majority (85%-97%) of the SMCs in the lung at E18.5 as well as alveolar MYF progenitors in the lung parenchyma. Fluorescence-activated cell sorting-based isolation of different subpopulations of ACTA2(+) lineage-traced cells followed by gene arrays, identified transcriptomic signatures for alveolar MYF progenitors versus airway and vascular SMCs at E18.5. Our results establish a new transcriptional landscape for further experiments addressing the function of signaling pathways in the formation of different subpopulations of ACTA2(+) cells.