GSE171745
Experimental and clinical assessment of angiogenesis-related parameters inpatients with juvenile idiopathic arthritis-associated uveitis
Introduction: In chronic and treatment-resistant cases of juvenile idiopathic arthritis-associated uveitis (JIAU), alterations of the iris vasculature may be observed clinically. We sought to investigate this phenomenon by combining clinical and laboratory analyses.  Methods: Iris tissue (IT), aqueous humor (AH) and serum samples from patients with clinically inactive JIAU (n=30) or acute anterior uveitis (AAU; n=18), and primary open angle glaucoma (POAG; n=20) were obtained during ocular surgery. Samples were analyzed by RNA-sequencing (RNA-Seq), qRT-PCR, liquid chromatography, high–definition mass spectrometry technique (LC-MS), Western-Blot (WB), and LEGENDplex? analysis. Pattern of iris vasculature in JIAU patients was assessed qualitatively via fluorescein and indocyanine green angiography (FLA/ICGA).  Results: RNA-Seq of IT showed significantly differential expression (DE) of 136 genes between JIAU and POAG, of which 15 were associated with angiogenesis. qRT-PCR, performed to validate RNA-Seq results, showed significant upregulation of the angiogenesis-related genes Kdr, Angpt-1, Tie-1, Tie-2 and Mmrn2 in IT (JIAU vs POAG). LC-MS of IT revealed a total number of 56 DE proteins (JIAU vs POAG), of which asporin, lumican and decorin were associated with angiogenesis and showed increased (p>0.05) expression on WB analysis. LEGENDplex? analysis showed upregulation of ANGPT-2 in AH from JIAU compared to AAU and POAG, whereas VEGF was upregulated in AAU. Iris vascular leakage, hypoperfusion and neovascularization were observed by FLA/ICGA in JIAU patients with treatment-refractory complicated course of uveitis.  Conclusion: Angiogenesis-related genes and proteins are upregulated in the iris tissue of JIAU patients, correlating with increased vascular permeability, hypoperfusion, and neovascularization of the anterior uveal tissue in chronic and complicated JIAU. Overall design: JIAU (study group): 4 samples; POAG (control group): 4 samples