The genome-wide transcriptional response to neonatal hyperoxia identifies Ahr as a key regulator
The genome-wide transcriptional response to neonatal hyperoxia identifies Ahr as a key regulator
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DOI：
10.1152/ajplung.00200.2014
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发表时间：
2014-10-01
影响因子：
4.9
通讯作者：
O'Reilly, Michael A.
中科院分区：
文献类型：
--
作者：
Bhattacharya, Soumyaroop;Zhou, Zhongyang;O'Reilly, Michael A.
需要补充氧气的早产儿患支气管肺发育不良（BPD）的风险增加。涉及新生儿暴露于过量氧气浓度（高氧）的啮齿动物模型有助于确定 BPD 相关病理机制。对新生小鼠肺部高氧影响的全基因组评估可以识别新的 BPD 相关基因和通路。新生 C57BL/6 小鼠暴露于 100% 氧气中 10 天，并使用全肺组织 RNA 进行高通量、基于测序的转录组分析 (RNA-Seq)。微阵列的显着性分析和独创性路径分析用于识别受影响的基因和路径。通过 qPCR 验证所选基因的表达模式。在培养的小鼠肺上皮细胞中进一步测试了基因之间的机制关系。我们确定了 300 个基因在新生儿急性高氧血症后受到显着和实质性影响。高氧肺中失调的典型途径包括核因子（红细胞衍生 2）样 2 介导的氧化应激信号、p53 信号、eNOS 信号和芳烃受体 (Ahr) 途径。聚类分析确定 Ccnd1、Cdkn1a 和 Ahr 是高氧反应中的关键调节节点，其中 Ahr 是主要效应节点。在肺上皮细胞中评估了 Ahr 的机制作用，我们证实了其调节多种高氧标记物表达的能力，包括 Cdkn1a、Pdgfrb 和 A2m。我们得出的结论是，对 BPD 样病理的急性新生儿高氧模型中的基因调控进行的全球评估已确定 Ahr 是基因失调的驱动因素之一。
Premature infants requiring supplemental oxygen are at increased risk for developing bronchopulmonary dysplasia (BPD). Rodent models involving neonatal exposure to excessive oxygen concentrations (hyperoxia) have helped to identify mechanisms of BPD-associated pathology. Genome-wide assessments of the effects of hyperoxia in neonatal mouse lungs could identify novel BPD-related genes and pathways. Newborn C57BL/6 mice were exposed to 100% oxygen for 10 days, and whole lung tissue RNA was used for high-throughput, sequencing-based transcriptomic analysis (RNA-Seq). Significance Analysis of Microarrays and Ingenuity Pathway Analysis were used to identify genes and pathways affected. Expression patterns for selected genes were validated by qPCR. Mechanistic relationships between genes were further tested in cultured mouse lung epithelial cells. We identified 300 genes significantly and substantially affected following acute neonatal hyperoxia. Canonical pathways dysregulated in hyperoxia lungs included nuclear fctor (erythryoid-derived-2)-like 2-mediated oxidative stress signaling, p53 signaling, eNOS signaling, and aryl hydrocarbon receptor (Ahr) pathways. Cluster analysis identified Ccnd1, Cdkn1a, and Ahr as critical regulatory nodes in the response to hyperoxia, with Ahr serving as the major effector node. A mechanistic role for Ahr was assessed in lung epithelial cells, and we confirmed its ability to regulate the expression of multiple hyperoxia markers, including Cdkn1a, Pdgfrb, and A2m. We conclude that a global assessment of gene regulation in the acute neonatal hyperoxia model of BPD-like pathology has identified Ahr as one driver of gene dysregulation.