Abstract
Purpose :
To uncover molecules of CsA & CS resistance and the mechanism whereby CLB overcomes the resistance.
Methods :
RNA sequencing (RNA-seq) based transcriptomics and isobaric tags for relative and absolute quantification (iTRAQ) based proteomics were performed to screen potential resistant molecules in VKH disease. Gain- and loss-of-function assays in vitro were carried out to assess the resistant effect of RPS4Y1 on CsA & CS. A rescue experiment was used to identify the improved effect of CLB on the CsA & CS resistance by RPS4Y1 suppression.
Results :
A total of 1,697 differentially expressed genes (DEGs) and 21 differentially expressed proteins (DEPs) were screened by transcriptomics and proteomics in CD4+ T cells between CsA & CS-resistant and -sensitive VKH patients. RPS4Y1 and HLA-DQA1 were selected as candidate genes after integrative omics analysis. Quantitative real time PCR (RT-qPCR) and Parallel Reaction Monitoring (PRM) demonstrated that RPS4Y1 was up-regulated at the mRNA and protein level in CsA & CS-resistant VKH patients. Gain-and loss-of-function assays showed that RPS4Y1 regulated the resistance of CD4+ T cells from male VKH patients to CsA & CS. Importantly, A rescue experiment indicated that CLB could reverse the resistance by suppressing RPS4Y1 expression.
Conclusions :
The present study identified RPS4Y1 as an important CsA & CS resistant gene and showed that CLB could counteract CsA & CS resistance by RPS4Y1 suppression.
This is a 2020 ARVO Annual Meeting abstract.