DataSheet_1_Metabolomic Analysis of Aqueous Humor Identifies Aberrant Amino Acid and Fatty Acid Metabolism in Vogt-Koyanagi-Harada and Behcet’s Disease.doc

To investigate aqueous metabolic profiles in Vogt-Koyanagi-Harada (VKH) and Behcet’s disease (BD), 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 MetaboAnalyst. 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 CoA 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-tRNA biosynthesis was altered in both VKH and BD when compared to controls. 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 BD and VKH.

为探讨伏格特-小柳-原田综合征（Vogt-Koyanagi-Harada, VKH）与贝赫切特病（Behcet’s disease, BD）患者的房水代谢谱，我们采用超高效液相色谱-四极杆飞行时间质谱联用技术，对上述患者及健康对照者的房水样本进行检测。采用单变量logistic回归分析三组样本的代谢物水平，并将筛选得到的差异代谢物通过MetaboAnalyst进行后续通路分析。结果显示，偏最小二乘判别分析（partial-least squares discrimination analysis）与层次聚类分析（hierarchical clustering analysis）均可在对比VKH、BD患者与健康对照时呈现特异性的房水代谢谱特征。VKH患者与健康对照相比共存在28种差异代谢物，BD患者与健康对照相比则有29种差异代谢物。氨基酸与脂肪酸为差异代谢物中丰度最高的两类代谢物。进一步的通路富集分析表明，VKH患者与健康对照相比存在泛酸与辅酶A（CoA）生物合成通路异常；BD患者与健康对照相比则存在D-精氨酸与D-鸟氨酸代谢、苯丙氨酸代谢通路异常。氨基酰-tRNA生物合成通路在VKH与BD患者与健康对照的对比中均出现异常改变。本研究结果提示，氨基酸代谢以及棕榈酸、油酸这两种脂肪酸或参与BD与VKH的发病过程。