银屑病发生发展与转归过程中血浆胞外囊泡分子标记研究

Psoriasis is no longer regard as a monolithic entity, but rather as several distinct entities within the spectrum of “psoriasis.” Psoriasis is a heterogeneous disease whose severity depends on the total skin surface affected, the specific areas affected, the severity of scaling and erythema and the comorbidities, including cardiovascular disease, diabetes, hypertension, metabolism syndrome and arthritis. Biomarkers would help us understand more quickly how patients are responding to new medications that are under development for this disease. Extracellular vesicles (EV) have emerged as important mediators of intercellular communication, being involved in the transmission of biological signals between cells to regulate a diverse range of biological processes. Functions proposed for EVs include transfer of drug resistance, growth regulation, quorum sensing, immune regulation, developmental modulation, and neurotransmission. At present, at least three main types of EV are recognized according to an evolving consensus nomenclature: exosomes, microvesicles (MVs; also called ectosomes or microparticles), and apoptotic bodies (ABs). EV-mediated signals can be transmitted by all the different biomolecule categories – protein, lipids, nucleic acids and sugars – and the unique package of this information provides both protection and the option of simultaneous delivery of multiple different messengers even to sites remote to the vesicular origin. Psoriasis is a state of systemic inflammation, because many inflammation biomarkers are detected in the blood of patients with psoriasis and are associated with disease activity. Therefore, we hypotheses that molecular profiling of various EV types in psoriasis may be unique molecular signatures that differentially modulate cellular functions and may be used for development, prognosis, diagnosis, and, possibly, directed therapy of psoriasis. The aim of the present study was to validate whether EVs specifically accumulate in the sera of a large heterogeneous cohort of male and female psoriasis patients compared to healthy controls and patients with other immune disorders including eczema. We also assessed whether EVs levels would be reduced in sera from psoriasis patients with immunosuppressive medication, compared to psoriasis patients without immunosuppressants, and thus if circulating EV levels would be responsive to immunomodulation. Investigation on the molecular pathways that explain the relationship among psoriasis, obesity, diabetes, and the metabolic syndrome. By solving these and related questions, we anticipate that more extensive research to EV research will help clarify the complex biology of psoriasis and contribute to the design of improved diagnostics and therapies.

银屑病不是单一的独立病种，而是具有不同阶段的慢性炎症性系统性病谱性疾病，病情的严重程度取决于皮损面积、部位及红斑鳞屑的严重程度和合并症，包括心血管疾病、糖尿病、高血压、代谢综合征和关节疼痛等。研究发现细胞外囊泡对细胞间信号调节具有重要作用，参与调节多种生物反应过程。胞外囊泡分为外泌体、微囊泡和凋亡小体，运载的信号多种多样，包括蛋白质、脂质、核酸及糖类等，可作为反映机体健康及疾病的生物标记。而银屑病患者血中存在大量与疾病活动相关的炎症标记分子，因此我们的假说：胞外囊泡中的某些分子，可作为银屑病发生发展和转归的标记，以及银屑病的诊断和治疗标记物。因此，我们拟通过比较大样本健康人群、其他免疫性疾病（包括湿疹）与不同类型、不同阶段、不同合并症银屑病患者以及治疗前后血清中胞外囊泡的差异，确立银屑病中特异性表达的、与银屑病发生、发展和转归相关的特异性胞外囊泡成分，阐明银屑病与合并症之间的发病关联机制。

银屑病是一种免疫介导的慢性炎症性皮肤病，越来越多的证据表明胞外囊泡中的microRNAs（miRNAs）在免疫性疾病中发挥重要作用。因此，我们采用RNA高通量测序技术探究了银屑病患者血清胞外囊泡中的miRNAs表达谱。结果显示，与健康对照相比，在银屑病患者血清胞外囊泡中有50种miRNAs差异表达，其中26种miRNAs上调，24种miRNAs下调。经qRT-PCR进一步验证发现，银屑病患者血清胞外囊泡中的miR-199a-3p表达水平较健康对照及玫瑰糠疹患者显著升高，并具有较高的受试者工作特征曲线。同时，miR-199a-3p的表达水平与银屑病面积和严重程度指数、体表受累面积呈正相关。此外，银屑病患者经系统性治疗达PASI 90后，血清胞外囊泡中的miR-199a-3p显著降低。有趣的是，在玫瑰糠疹患者的血清胞外囊泡中，miR-500a-3p、miR-484、miR-185-5p、miR-27a-5p和let-7f-5p的表达水平显著高于银屑病患者和健康对照，而miR-1255b-5p则降低，这些差异表达的miRNAs均具有较好的受试者工作特征曲线，具有潜在的诊断价值。生物信息学分析预测发现了miR-199a-3p的114个mRNA靶标，它们主要富集于局部粘附途径和mTOR途径。这项研究显示，银屑病、玫瑰糠疹患者的血清胞外囊泡中具有独特的miRNA表达谱，这些差异表达的miRNAs不仅可以作为临床诊断和疾病评估的潜在生物标志物，而且有望成为发病机制研究的靶分子。