Department of Cell Biology, University of Oklahoma Health science Center, Oklahoma City, Oklahoma, 73104, USA.
Department of Cellular Biology & Anatomy, Augusta University, Augusta, GA, 30912, United States.
Sci Rep. 2019 Aug 29;9(1):12567. doi: 10.1038/s41598-019-48983-8.
Keratoconus (KC) is a multi-factorial corneal ectasia with unknown etiology affecting approximately 1:2000 people worldwide. Dysregulated gene expression, using RNA-Seq technology, have been reported in KC corneal tissue. However, the differential expression of genes, in KC corneal stromal cells have been widely ignored. We utilized mRNA-Seq to analyze gene expression in primary human corneal stromal cells derived from five non-Keratoconus healthy (HCF) and four Keratoconus (HKC) donors. Selected genes were further validated using real time PCR (RT-PCR). We have identified 423 differentially expressed genes with 187 down- and 236 up-regulated in KC-affected corneal stromal cells. Gene ontology analysis using WebGestalt indicates the enrichment of genes involved in cell migration, extracellular matrix, adherens junction, and MAPK signaling. Our protein-protein interaction network analysis identified several network seeds, such as EGFR, NEDD4, SNTA1, LGALS3BP, HSPB1, SDC2, MME, and HIF1A. Our work provides an otherwise unknown information on the transcriptional changes in HKCs, and reveals critical mechanisms of the cellular compartment. It also highlights the importance of human-based in vitro studies on a disease that currently lacks strong biomarkers and animal models.
圆锥角膜（KC）是一种多因素的角膜扩张疾病，病因不明，全球约有 1/2000 的人受到影响。使用 RNA-Seq 技术已经报道了 KC 角膜组织中基因表达的失调。然而，KC 角膜基质细胞中基因的差异表达却被广泛忽视。我们利用 mRNA-Seq 技术分析了来自 5 名非圆锥角膜健康（HCF）和 4 名圆锥角膜（HKC）供体的原代人角膜基质细胞中的基因表达。选择的基因进一步通过实时 PCR（RT-PCR）进行验证。我们鉴定了 423 个差异表达基因，其中 187 个下调和 236 个上调在 KC 相关的角膜基质细胞中。使用 WebGestalt 进行基因本体论分析表明，富集了参与细胞迁移、细胞外基质、黏着连接和 MAPK 信号通路的基因。我们的蛋白质-蛋白质相互作用网络分析确定了几个网络种子，如 EGFR、NEDD4、SNTA1、LGALS3BP、HSPB1、SDC2、MME 和 HIF1A。我们的工作提供了关于 HKC 中转录变化的未知信息，并揭示了细胞区室的关键机制。它还强调了在目前缺乏强有力的生物标志物和动物模型的疾病中进行基于人类的体外研究的重要性。
