Klf9 is a key feedforward regulator of the transcriptomic response to glucocorticoid receptor activity
Klf9 is a key feedforward regulator of the transcriptomic response to glucocorticoid receptor activity
复制标题
DOI：
10.1038/s41598-020-68040-z
复制
发表时间：
2020-07-10
影响因子：
4.6
通讯作者：
Coffman, James A.
中科院分区：
文献类型：
--
作者：
Gans, Ian;Hartig, Ellen, I;Coffman, James A.
斑马鱼最近成为研究糖皮质激素信号传导的发育作用和糖皮质激素诱导的发育编程机制的模型系统。为了评估糖皮质激素受体（GR）在这种编程中的作用，我们使用CRISPR-Cas9产生了一种新的移码突变GR（369-），它消除了DNA结合结构域上游所有潜在的框内起始密码子。使用RNA-seq询问这种突变如何影响正常条件下和慢性皮质醇治疗下的幼虫转录组，我们发现GR介导了治疗的大部分效果，矛盾的是，皮质醇治疗的幼虫的转录组更像缺乏GR的幼虫，而不是GR的幼虫，这表明皮质醇治疗的幼虫产生了GR抗性。在GR（369-）幼虫中低表达和在皮质醇处理的幼虫中一贯过表达的一个转录调节因子是klf 9。因此，我们使用CRISPR-Cas9介导的klf 9突变和RNA-seq来评估正常和皮质醇处理的幼虫中Klf 9依赖性基因的表达。我们的研究结果表明Klf 9对慢性皮质醇暴露的转录组学反应有显著贡献，介导了我们先前报道的促炎基因的上调。
The zebrafish has recently emerged as a model system for investigating the developmental roles of glucocorticoid signaling and the mechanisms underlying glucocorticoid-induced developmental programming. To assess the role of the Glucocorticoid Receptor (GR) in such programming, we used CRISPR-Cas9 to produce a new frameshift mutation, GR(369-), which eliminates all potential in-frame initiation codons upstream of the DNA binding domain. Using RNA-seq to ask how this mutation affects the larval transcriptome under both normal conditions and with chronic cortisol treatment, we find that GR mediates most of the effects of the treatment, and paradoxically, that the transcriptome of cortisol-treated larvae is more like that of larvae lacking a GR than that of larvae with a GR, suggesting that the cortisol-treated larvae develop GR resistance. The one transcriptional regulator that was both underexpressed in GR(369-) larvae and consistently overexpressed in cortisol-treated larvae was klf9. We therefore used CRISPR-Cas9-mediated mutation of klf9 and RNA-seq to assess Klf9-dependent gene expression in both normal and cortisol-treated larvae. Our results indicate that Klf9 contributes significantly to the transcriptomic response to chronic cortisol exposure, mediating the upregulation of proinflammatory genes that we reported previously.