Table1_Lipidic Profile Changes in Exosomes and Microvesicles Derived From Plasma of Monoclonal Antibody-Treated Psoriatic Patients.DOCX

Psoriasis is a chronic immune-mediated inflammatory skin disorder affecting children and adults. To date no approved biomarkers for diagnosis of this disease and follow up of patients have been translated into clinical practice. Recently, extracellular vesicles (EVs) secreted by all cells and present in almost all biological fluids are playing a crucial role in diagnosis and follow up of several diseases, including psoriasis. Since many psoriatic patients show altered plasma lipid profiles and since EVs have been involved in psoriasis pathogenesis, we studied the phospholipid profile of EVs, both microvesicles (MV) or exosomes (Exo), derived from plasma of psoriatic patients undergoing systemic biological treatment (secukinumab, ustekinumab, adalimumab), in comparison with EVs of untreated patients and healthy donors (HD). EVs were evaluated by immune electronmicroscopy for their morphology and by NanoSight for their amount and dimensions. EV phospholipid profiling was performed by High Resolution Liquid Chromatography-Mass Spectrometry and statistical Partial Least Squares Discriminant Analysis. Our results demonstrated that psoriatic patients showed a higher concentration of both MV and Exo in comparison to EVs from HD. The phospholipid profile of Exo from psoriatic patients showed increased levels of phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylglycerol and lysoPC compared to Exo from HD. Sphingomyelin (SM) and phosphatidylinositol (PI) are the only phospholipid classes whose levels changed in MV. Moreover, the therapy with ustekinumab seemed to revert the PE and PC lipid composition of circulating Exo towards that of HD and it is the only one of the three biological drugs that did not alter SM expression in MV. Therefore, the determination of lipid alterations of circulating EVs could harbor useful information for the diagnosis and drug response in psoriatic patients.

银屑病（Psoriasis）是一种累及儿童与成人的慢性免疫介导性炎症性皮肤疾病。截至目前，尚无经批准的该疾病诊断与患者随访用生物标志物转化应用于临床实践。近年来，由所有细胞分泌、几乎存在于所有生物体液中的细胞外囊泡（extracellular vesicles, EVs）在包括银屑病在内的多种疾病的诊断与随访中发挥关键作用。鉴于诸多银屑病患者存在血浆脂质谱异常，且细胞外囊泡已被证实参与银屑病发病机制，本研究针对接受系统性生物治疗（司库奇尤单抗（secukinumab）、乌司奴单抗（ustekinumab）、阿达木单抗（adalimumab））的银屑病患者血浆来源的微囊泡（microvesicles, MV）与外泌体（exosomes, Exo）的磷脂谱进行了分析，并与未接受治疗的银屑病患者及健康受试者（healthy donors, HD）的细胞外囊泡进行对照。
研究通过免疫电子显微镜评估细胞外囊泡的形态，借助NanoSight检测其浓度与粒径尺寸。采用高分辨液相色谱-质谱联用法完成细胞外囊泡的磷脂谱分析，并运用偏最小二乘判别分析进行统计学检验。
研究结果显示，与健康受试者的细胞外囊泡相比，银屑病患者的微囊泡与外泌体浓度均显著更高。银屑病患者外泌体的磷脂谱中，磷脂酰胆碱（phosphatidylcholine, PC）、磷脂酰乙醇胺（phosphatidylethanolamine, PE）、磷脂酰甘油及溶血磷脂酰胆碱（lysoPC）的水平较健康受试者明显升高。仅鞘磷脂（sphingomyelin, SM）与磷脂酰肌醇（phosphatidylinositol, PI）这两类磷脂在微囊泡中出现水平变化。此外，乌司奴单抗治疗似乎可将循环中外泌体的磷脂酰乙醇胺与磷脂酰胆碱的脂质组成恢复至健康受试者水平，且是三种生物制剂中唯一未改变微囊泡中鞘磷脂表达的药物。因此，检测循环细胞外囊泡的脂质异常，可为银屑病患者的诊断及药物响应评估提供有价值的参考信息。