RNA-Seq analysis reveals gene expression changes induced by IL-6 trans-signaling activation in retinal endothelial cells

Cytokine. 2021 Mar:139:155375. doi: 10.1016/j.cyto.2020.155375. Epub 2020 Dec 28.


Background: Increasing evidence suggests that interleukin-6 (IL-6) trans-signaling plays a critical role in the pathogenesis of diabetic retinopathy (DR). We have previously shown that activation of IL-6 trans-signaling induces barrier dysfunction in human retinal endothelial cells (HRECs). However, the molecular mechanisms underlying these effects are not clear. The purpose of this study was to discover global gene expression changes in HRECs following activation of IL-6 trans-signaling.

Methods: HRECs were treated with IL-6 and soluble IL-6R to activate IL-6 trans-signaling, and sgp130Fc treatment was used for IL-6 trans-signaling inhibition. RNA-Seq analyses were performed for global gene expression profiling. Differential expression was determined using DESeq2, and bioinformatic analyses were performed to associate the differentially expressed genes with biological functions and pathways.

Results: Our analyses revealed 445 differentially expressed genes (318 upregulated and 127 downregulated) in HRECs after IL-6 trans-signaling activation. We identified several novel genes not previously associated with IL-6 signaling or endothelial dysfunction. Leukocyte adhesion, diapedesis and chemokine signaling pathways are highly enriched in differentially expressed genes. Inhibition of IL-6 trans-signaling with sgp130Fc abrogated these changes, thus highlighting the therapeutic potential of this drug.

Conclusions: This study identified significant gene expression changes caused by IL-6 trans-signaling activation in HRECs. Identification of such changes has the potential to uncover the precise molecular mechanisms of IL-6 trans-signaling mediated effects in the pathology of DR.

Keywords: Bioinformatic analyses; Endothelial cells; Gene expression; IL-6 trans-signaling; RNA-Seq; sgp130Fc.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Cells, Cultured
  • Down-Regulation / genetics
  • Endothelial Cells / metabolism*
  • Gene Expression / genetics*
  • Gene Expression Profiling / methods
  • Humans
  • Interleukin-6 / genetics*
  • RNA-Seq / methods
  • Retina / metabolism*
  • Signal Transduction / genetics*
  • Up-Regulation / genetics


  • IL6 protein, human
  • Interleukin-6