Genome-wide retinal transcriptome analysis of endotoxin-induced uveitis in mice with next-generation sequencing

Mol Vis. 2017 Jul 3:23:395-406. eCollection 2017.

Abstract

Purpose: Endotoxin-induced uveitis (EIU) is a well-established mouse model for studying human acute inflammatory uveitis. The purpose of this study is to investigate the genome-wide retinal transcriptome profile of EIU.

Methods: The anterior segment of the mice was examined with a slit-lamp, and clinical scores were evaluated simultaneously. The histological changes in the posterior segment of the eyes were evaluated with hematoxylin and eosin (H&E) staining. A high throughput RNA sequencing (RNA-seq) strategy using the Illumina Hiseq 2500 platform was applied to characterize the retinal transcriptome profile from lipopolysaccharide (LPS)-treated and untreated mice. The validation of the differentially expressed genes (DEGs) was analyzed with real-time PCR.

Results: At the 24th hour after challenge, the clinical score of the LPS group was significantly higher (3.83±0.75, mean ± standard deviation [SD]) than that of the control group (0.08±0.20, mean ± SD; p<0.001). The histological evaluation showed a large number of inflammatory cells infiltrated into the vitreous cavity in the LPS group compared with the control group. A total of 478 DEGs were identified with RNA-seq. Among these genes, 406 were upregulated and 72 were downregulated in the LPS group. Gene Ontology (GO) enrichment showed three significantly enriched upregulated terms. Twenty-one upregulated and seven downregulated pathways were remarkably enriched by Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment. Eleven inflammatory response-, complement system-, fibrinolytic system-, and cell stress-related genes were validated to show similar results as the RNA-seq.

Conclusions: We first reported the retinal transcriptome profile of the EIU mouse with RNA-seq. The results indicate that the abnormal changes in the inflammatory response-, complement system-, fibrinolytic system-, and cell stress-related genes occurred concurrently in EIU. These genes may play an important role in the pathogenesis of EIU. This study will lead to a better understanding of the underlying mechanisms and shed light on discovering novel therapeutic targets for ocular inflammation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Anterior Chamber / metabolism
  • Anterior Chamber / pathology
  • Cluster Analysis
  • Down-Regulation / genetics
  • Endotoxins
  • Female
  • Gene Expression Profiling*
  • Gene Ontology
  • Genome*
  • High-Throughput Nucleotide Sequencing*
  • Inflammation / pathology
  • Lipopolysaccharides
  • Mice, Inbred BALB C
  • Real-Time Polymerase Chain Reaction
  • Reproducibility of Results
  • Retina / metabolism*
  • Retina / pathology*
  • Up-Regulation / genetics
  • Uveitis / genetics*
  • Uveitis / pathology

Substances

  • Endotoxins
  • Lipopolysaccharides