Pericyte-like cells undergo transcriptional reprogramming and distinct functional adaptations in acute lung injury.
Pericyte-like cells undergo transcriptional reprogramming and distinct functional adaptations in acute lung injury.
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DOI：
10.1096/fj.201903192rr
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发表时间：
2021-04
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我们之前报道了周细胞样细胞在肺损伤中作为功能性前哨免疫细胞的作用。然而，关于周细胞在肺损伤中的生物学作用仍不清楚。由于肺周细胞样细胞在肺泡内的解剖位置，它们可以很好地感知上皮屏障的破坏并协调局部炎症反应。在本报告中，我们描述了急性肺损伤（ALI）小鼠模型中受损和未受损肺的肺泡成分激活后周细胞样细胞的转录反应和功能变化。使用PDGFRβ作为选择标记，从肺消化液中纯化的周细胞样细胞，如前所述在培养中扩增。我们用重组人Fas配体（rhFasL）灌注机械通气诱导小鼠无菌急性肺损伤。然后，我们从受伤和未受伤的小鼠中收集支气管肺泡灌洗液（BALF）。将纯化的周细胞样细胞仅暴露于生长培养基（对照组）、未损伤小鼠的BALF和损伤小鼠的BALF中6和24小时。从这些处理条件中收集的RNA被处理为RNAseq。通过途径分析确定的感兴趣的靶点通过体外和体内试验进行验证。我们观察到，在未损伤和损伤的BALF处理后6小时，周细胞样细胞的整体转录变化强劲，但这种反应仅在暴露于损伤的BALF后持续24小时。损伤BALF处理的周细胞功能富集分析显示，促炎、细胞迁移和血管生成相关通路被激活，而与组织发育和细胞分化相关的过程被下调。我们在体外和体内使用功能生物学试验验证了炎症、血管生成和细胞迁移途径中选择的上调靶点。我们得出结论，肺周细胞样细胞对未损伤和损伤肺的肺泡室内容物均有高度反应，但损伤的BALF引起更持久的反应。激活的周细胞样细胞所表现出的炎症、血管生成和迁移变化强调了这些特化基质细胞在急性肺损伤背景下的表型可塑性。
We previously reported on the role of pericyte-like cells as functional sentinel immune cells in lung injury. However, much about the biological role of pericytes in lung injury remains unknown. Lung pericyte-like cells are well-positioned to sense disruption to the epithelial barrier and coordinate local inflammatory responses due to their anatomic niche within the alveoli. In this report, we characterized transcriptional responses and functional changes in pericyte-like cells following activation by alveolar components from injured and uninjured lungs in a mouse model of acute lung injury (ALI). Purified pericyte-like cells from lung digests using PDGFRβ as a selection marker were expanded in culture as previously described. We induced sterile acute lung injury in mice with recombinant human Fas ligand (rhFasL) instillation followed by mechanical ventilation. We then collected bronchoalveolar lavage fluid (BALF) from injured and uninjured mice. Purified pericyte-like cells in culture were exposed to growth media only (control), BALF from uninjured mice, and BALF from injured mice for 6 and 24 h. RNA collected from these treatment conditions were processed for RNAseq. Targets of interest identified by pathway analysis were validated using in vitro and in vivo assays. We observed robust global transcriptional changes in pericyte-like cells following treatment with uninjured and injured BALF at 6 h, but this response persisted for 24 h only after exposure to injured BALF. Functional enrichment analysis of pericytes treated with injured BALF revealed activation of pro-inflammatory, cell migration and angiogenesis-related pathways, whereas processes associated with tissue development and cell differentiation were down-regulated. We validated select upregulated targets in the inflammatory, angiogenic, and cell migratory pathways using functional biological assays in vitro and in vivo. We conclude that lung pericyte-like cells are highly responsive to alveolar compartment content from both uninjured and injured lungs, but injured BALF elicits a more sustained response. The inflammatory, angiogenic, and migratory changes exhibited by activated pericyte-like cells underscore the phenotypic plasticity of these specialized stromal cells in the setting of acute lung injury.