Department of Medical Genetics, University of Wisconsin, Madison, Wisconsin;
Physiol Genomics. 2014 Apr 15;46(8):277-89. doi: 10.1152/physiolgenomics.00126.2013. Epub 2014 Feb 18.
Increased angiogenesis, inflammation, and proliferation are hallmarks of diseased tissues, and in vivo models of these disease phenotypes can provide insight into disease pathology. Dstn(corn1) mice, deficient for the actin depolymerizing factor destrin (DSTN), display an increase of serum response factor (SRF) that results in epithelial hyperproliferation, inflammation, and neovascularization in the cornea. Previous work demonstrated that conditional ablation of Srf from the corneal epithelium of Dstn(corn1) mice returns the cornea to a wild-type (WT) like state. This result implicated SRF as a major regulator of genes that contributes to abnormal phenotypes in Dstn(corn1) cornea. The purpose of this study is to identify gene networks that are affected by increased expression of Srf in the Dstn(corn1) cornea. Microarray analysis led to characterization of gene expression changes that occur when conditional knockout of Srf rescues mutant phenotypes in the cornea of Dstn(corn1) mice. Comparison of gene expression values from WT, Dstn(corn1) mutant, and Dstn(corn1) rescued cornea identified >400 differentially expressed genes that are downstream from SRF. Srf ablation had a significant effect on genes associated with epithelial cell-cell junctions and regulation of actin dynamics. The majority of genes affected by SRF are downregulated in the Dstn(corn1) mutant cornea, suggesting that increased SRF negatively affects transcription of SRF gene targets. ChIP-seq analysis on Dstn(corn1) mutant and WT tissue revealed that, despite being present in higher abundance, SRF binding is significantly decreased in the Dstn(corn1) mutant cornea. This study uses a unique model combining genetic and genomic approaches to identify genes that are regulated by SRF. These findings expand current understanding of the role of SRF in both normal and abnormal tissue homeostasis.
血管生成、炎症和增殖增加是病变组织的特征,这些疾病表型的体内模型可以深入了解疾病的病理。缺乏肌动蛋白解聚因子 destrin (DSTN)的 Dstn(corn1)小鼠表现出血清反应因子 (SRF)的增加,导致角膜上皮过度增殖、炎症和新生血管形成。先前的工作表明,条件性敲除 Dstn(corn1)小鼠角膜上皮中的 Srf 可使角膜恢复到类似于野生型 (WT)的状态。这一结果表明 SRF 是主要调节因子之一,它有助于 Dstn(corn1)角膜的异常表型。本研究的目的是鉴定受 Dstn(corn1)角膜中 SRF 表达增加影响的基因网络。微阵列分析导致了特征描述,即在 Dstn(corn1)小鼠角膜中条件性敲除 Srf 可挽救突变表型时发生的基因表达变化。WT、Dstn(corn1)突变体和 Dstn(corn1)挽救型角膜的基因表达值比较鉴定出了 >400 个受 SRF 下游影响的差异表达基因。Srf 敲除对与上皮细胞-细胞连接和肌动蛋白动力学调节相关的基因有显著影响。受 SRF 影响的大多数基因在 Dstn(corn1)突变体角膜中下调,这表明增加的 SRF 负调控 SRF 基因靶标的转录。对 Dstn(corn1)突变体和 WT 组织进行 ChIP-seq 分析表明,尽管 SRF 结合的丰度较高,但在 Dstn(corn1)突变体角膜中,SRF 结合显著减少。本研究使用一种独特的模型,结合遗传和基因组方法来鉴定受 SRF 调节的基因。这些发现扩展了当前对 SRF 在正常和异常组织稳态中的作用的理解。