Galderma R&D, 2400, route des colles, 06410, Biot, France.
132 rue d'Assas 75006, Paris, France.
Br J Dermatol. 2017 Aug;177(2):470-488. doi: 10.1111/bjd.15346. Epub 2017 May 18.
Protein expression is disturbed in the psoriatic stratum corneum (SC). Noninvasive methods for the description of pathophysiological changes and drug profiling in psoriasis are desirable.
Undertake large-scale noninvasive protein expression studies in psoriatic SC to identify biomarkers of pathophysiological processes and use them for drug profiling.
Psoriatic SC was harvested through repetitive tape-stripping. Nonlesional and lesional SC, as well as vehicle-treated and drug-treated lesional SC samples were collected. Protein extracts from nonlesional and lesional skin biopsies were used for comparison. Calcipotriol-betamethasone (CB) was used as a reference medication. Proteins extracted from pooled tape strips were quantified using mass spectrometry (MS), Western blotting, enzyme-linked immunosorbent assay and Luminex technologies.
MS-based methods identified 140 proteins differentially expressed in psoriatic SC. Epidermis development, glycolysis, regulation of apoptosis, cytoskeleton organization and peptide cross-linking were modulated, all reflecting perturbed epidermal differentiation. Using antibody-based techniques, increased levels of sICAM1, of CXCL1- and CXCL8-attracting neutrophils, of CXCL10- and CCL4-attracting T helper (Th) 1 cells, and of CCL2- and CCL4-attracting monocytes and dendritic cells were observed. Quantification of the Th1 and Th17 markers tumour necrosis factor, interleukin (IL) 12B, IL17A and IL17F in lesional SC was successful, while the Th2 cytokines IL4, IL5 and IL13, not involved in the disease process, were not detected. The pruritic cytokine IL31 was detected in lesional SC. CXCL1, CXCL8, CXCL10 and sICAM were used to investigate disease remission, ranking three topical treatments according to their known clinical efficacy.
Protein biomarker quantification in psoriatic SC detects key pathophysiological mechanisms and enables noninvasive drug profiling in translational medicine settings.
银屑病患者表皮蛋白质表达紊乱。人们期望能找到一种非侵入性的方法来描述银屑病的病理生理变化和药物作用谱。
在银屑病表皮中进行大规模的非侵入性蛋白质表达研究，以鉴定病理生理过程的生物标志物，并用于药物作用谱分析。
通过重复胶带撕除法采集银屑病表皮。采集非皮损和皮损表皮，以及接受药物和赋形剂处理的皮损表皮。将非皮损和皮损皮肤活检的蛋白提取物进行对比。使用卡泊三醇倍他米松（CB）作为参考药物。使用质谱（MS）、Western blot、酶联免疫吸附测定和 Luminex 技术对从胶带中提取的蛋白质进行定量。
MS 方法鉴定出 140 种在银屑病表皮中差异表达的蛋白质。表皮发育、糖酵解、细胞凋亡调控、细胞骨架组织和肽交联被调节，这反映了表皮分化失调。使用抗体技术，观察到 sICAM1、吸引中性粒细胞的 CXCL1 和 CXCL8、吸引 Th1 细胞的 CXCL10 和 CCL4、吸引单核细胞和树突状细胞的 CCL2 和 CCL4 的水平升高。成功定量了皮损表皮中的 Th1 和 Th17 标志物肿瘤坏死因子、白细胞介素（IL）12B、IL17A 和 IL17F，而不参与疾病过程的 Th2 细胞因子 IL4、IL5 和 IL13 则没有检测到。在皮损表皮中检测到瘙痒细胞因子 IL31。CXCL1、CXCL8、CXCL10 和 sICAM 用于研究疾病缓解情况，根据其已知的临床疗效对三种局部治疗方法进行排序。
在银屑病表皮中进行蛋白质生物标志物定量分析可以检测到关键的病理生理机制，并能在转化医学背景下进行非侵入性药物作用谱分析。
