iTRAQ 定量蛋白质组学揭示了可预测甲氨蝶呤疗效的银屑病生物标志物/通路。

iTRAQ-based quantitative proteomics reveals biomarkers/pathways in psoriasis that can predict the efficacy of methotrexate.

机构信息

Department of Dermatology, Institute of Dermatology, Huashan Hospital, Fudan University, Shanghai, China.

CAS Key Laboratory of Receptor Research, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, China.

出版信息

J Eur Acad Dermatol Venereol. 2022 Oct;36(10):1784-1795. doi: 10.1111/jdv.18292. Epub 2022 Jun 24.

DOI:10.1111/jdv.18292

PMID:35666151

Abstract

BACKGROUND

Methotrexate (MTX) is the first-line medicine to treat psoriasis. So far, there has been less research on protein biomarkers to predict its efficacy by the proteomic technique.

OBJECTIVES

To evaluate differentially expressed proteins in peripheral mononuclear cells (PBMCs) between good responders (GRs) and non-responders (NRs) after MTX treatment, compared with normal controls (NCs).

METHODS

We quantified protein expression of PBMCs with four GRs and four NRs to MTX and four NCs by isobaric tags for relative and absolute quantification (iTRAQ), analysing and identifying proteins related to efficacy of MTX in 18 psoriatic patients.

RESULTS

A total of 3177 proteins had quantitative information, and 403 differentially expressed proteins (fold change ≥1.2, P < 0.05) were identified. Compared to NCs, upregulated proteins (ANXA6, RPS27A, EZR, XRCC6), participating in the activation of NF-κB, the JAK-STAT pathway and neutrophil degranulation were detected in GRs. The proteins (GPV, FN1, STOM), involving platelet activation, signalling and aggregation as well as neutrophil degranulation were significantly downregulated in GRs. These proteins returned to normal levels after MTX treatment. Furthermore, Western blotting identified the expression of ANXA6 and STAT1 in PBMCs, which were significantly downregulated in GRs, but not in NRs.

CONCLUSIONS

We identified seven differentially expressed and regulated proteins (ANXA6, GPV, FN1, XRCC6, STOM, RPS27A and EZR) as biomarkers to predict MTX efficacy in NF-κB signalling, JAK-STAT pathways, neutrophil degranulation, platelet activation, signalling and aggregation.

摘要

背景

甲氨蝶呤（MTX）是治疗银屑病的一线药物。迄今为止，利用蛋白质组学技术寻找预测其疗效的蛋白生物标志物的研究较少。

目的

评估甲氨蝶呤（MTX）治疗后外周血单个核细胞（PBMC）中对药物反应良好（GR）与反应不佳（NR）的患者与正常对照（NC）之间的差异表达蛋白。

方法

采用同位素标记相对和绝对定量（iTRAQ）技术，对 18 例银屑病患者的 4 例 GR 患者、4 例 NR 患者、4 例 NC 患者的 PBMC 蛋白表达进行定量分析，筛选与 MTX 疗效相关的蛋白。

结果

共获得 3177 个蛋白的定量信息，鉴定到 403 个差异表达蛋白（fold change≥1.2，P<0.05）。与 NC 相比，GR 中 NF-κB 激活、JAK-STAT 通路和嗜中性粒细胞脱颗粒相关的上调蛋白（ANXA6、RPS27A、EZR、XRCC6）被检测到。GR 中显著下调的蛋白（GPV、FN1、STOM），参与血小板激活、信号转导和聚集以及嗜中性粒细胞脱颗粒。经 MTX 治疗后，这些蛋白水平恢复正常。此外，Western blot 鉴定到 GR 中 ANXA6 和 STAT1 表达显著下调，但 NR 中未见下调。