Study On Identification Identification Of The Cyclosporin A Plus Corticosteroid Sensitive Molecule In Pediatric Uveitis And Aqueous Metabolomic Analysis In Uveitis

PART I STUDY ON IDENTIFICATION IDENTIFICATION OF THE CYCLOSPORIN A PLUS CORTICOSTEROID SENSITIVE MOLECULE IN PEDIATRIC IDIOPATHIC UVEITISBackground:Immature abilities on expression and sensation of children would lead to the delayed diagnosis and treatment of pediatric diseases.Pediatric idiopathic uveitis(PIU)is the most commonly seen pediatric uveitis entity.Combination of cyclosporin A(Cs A)with corticosteroids(CS)is widely used in the treatment of PIU.Although some PIU patients develop resistant to this combination regimen,most of them develop sensitive.However,the key sensitive molecule on this combination regimen remain unknown.Purpose:To investigate the molecules of Cs A&CS sensitive in PIU and the mechanism whereby.Methods:Firstly,transcriptomics and tandem mass tag(TMT)based proteomics were performed to investigate potential Cs A&CS sensitive molecules in PIU.Then RT-q PCR,PRM and WB were subjected to measure the m RNA and protein levels of the candidate sensitive molecules.Finally,loss of function experiments in vitro were conducted to assess the sensitive effect of B2M on Cs A&CS.Results:A total of 368 differentially expressed genes(DEGs)were detected by transcriptomics,and 101 differentially expressed proteins(DEPs)were detected by proteomics in CD4~+T cells from Cs A&CS sensitive and resistant PIU patients.Through integrative omics analysis,B2M,GGACT and ZNF236 were considered as three candidate molecules.The results of RT-q PCR,PRM and WB showed that B2M was up-regulated in Cs A&CS sensitive patients.In vitro,loss of function experiment showed that inhibition of B2M could increase the percentage of helper T(Th)1cells in sensitive patients and the IFN-γlevel.Conclusion:The current study identified B2M as an important Cs A&CS sensitive molecule for PIU,and it works through regulating Th1cells.PART II STUDY ON AQUEOUS METABOLOMIC ANALYSIS IN VOGT-KOYANAGI-HARADA AND BEHCET’S DISEASEBackground: Uveitis is known as one of the primary causes of blindness in the world.Vogt-Koyanagi-Harada(VKH)disease and Behcet’s disease(BD)are two of the most commonly seen uveitis entities in China.In view of the characteristics of recurrent and refractory of these two diseases,the study on the pathogenesis contributes to the prevention and treatment.Metabolomics can reflect local physiological events.Therefore,aqueous metabolomic analysis could advance our knowledgement about these two diseases.Purpose: To investigate aqueous metabolic profiles in VKH disease and BD.Methods: We applied ultra-high-performance liquid chromatography equipped with quadrupole time-of flight mass spectrometry in aqueous humor samples collected from these patients and controls.Metabolite levels in these three groups were analyzed by univariate logistic regression.The differential metabolites were subjected to subsequent pathway analysis by Metabo Analyst.Results: The results showed that both partial-least squares discrimination analysis and hierarchical clustering analysis showed specific aqueous metabolite profiles when comparing VKH,BD,and controls.There were 28 differential metabolites in VKH compared to controls and 29 differential metabolites in BD compared to controls.Amino acids and fatty acids were the two most abundant categories of differential metabolites.Furthermore,pathway enrichment analysis identified several perturbed pathways,including pantothenate and Co A biosynthesis when comparing VKH with the control group,and D-arginine and D-ornithine metabolism and phenylalanine metabolism when comparing BD with the control group.Aminoacyl-t RNA biosynthesis was altered in both VKH and BD when compared to controls.Conclusion: Our findings suggest that amino acids metabolism as well as two fatty acids,palmitic acid and oleic acid,may be involved in the pathogenesis of VKH disease and BD.