Department of Dermatology and Venereology, The First Affiliated Hospital, Anhui Medical University, Hefei, Anhui, China.
Institute of Dermatology, Anhui Medical University, Hefei, Anhui, China.
J Eur Acad Dermatol Venereol. 2021 Jun;35(6):1393-1406. doi: 10.1111/jdv.17112. Epub 2021 Jan 28.
Verrucous epidermal naevi (VEN) are benign skin tumours, considered keratinocytic epidermal naevi, that appear at birth or early childhood. VEN may display a range of appearances, depending on patient age. Although the number of studies regarding VEN is increasing, the exact mechanism of VEN is still unknown.
The aim of this study was to analyse the changes in the expression of protein factors in lesions of VEN children by TMT labelling-based quantitative proteomics.
A total of 8 children with VEN (5 for experiment and 3 for validation) and 8 healthy children (5 for experiment and 3 for validation) presented to the Department of Dermatology, the First Affiliated Hospital of Anhui Medical University, Boao Super Hospital, between January 2019 and November 2019. The lesions and lesion-adjacent tissues from children with VEN and naevus-adjacent normal skin tissues from children with pigmented naevi were defined as the VEN group, VENC group and C group, respectively. We performed a proteomics analysis to screen for differentially expressed proteins in the lesions of these individuals. We further performed Western blotting to validate the relative expression levels of nine targeted proteins in the validation group.
According to the proteomics results, a total of 4970 proteins were identified, and 4770 proteins were quantified. Among these proteins, 586 proteins were up- or downregulated at least 1.3-fold with a P-value < 0.05 (upregulated: 399, downregulated: 187) in lesions between the VEN group and the C group. These proteins played important roles in multiple biological functions, such as cornification, epidermal cell differentiation and neutrophil activation, and formed a complicated protein-protein interaction network. Of the 586 up- or downregulated proteins, nine were selected for further validation. According to Western blotting analysis results, the relative expression levels of Involucrin, NDUFA4, Loricrin, Keratin type II cytoskeletal 6A (Cytokeratin 6A), BRAF, Filaggrin, S100A7 and Desmocollin-3 were significantly upregulated in VEN children and may be associated with skin barrier dysfunction, epidermal cell overgrowth and differentiation, inflammation and immune and oxidative phosphorylation, which are involved in the pathogenesis of VEN.
According to TMT-based proteomics and Western blotting results, we identified eight noteworthy proteins, Involucrin, NDUFA4, Loricrin, Keratin type II cytoskeletal 6A, BRAF, Filaggrin, S100A7 and Desmocollin-3, that were upregulated in the lesions of VEN children and may be associated with the pathogenesis of VEN. Our findings provide new starting points for identifying precise pathogenic mechanisms or therapeutic targets for VEN.
疣状表皮痣（VEN）是一种良性皮肤肿瘤，被认为是角朊细胞表皮痣，在出生或幼儿期出现。VEN 可能表现出多种外观，具体取决于患者年龄。尽管关于 VEN 的研究越来越多，但 VEN 的确切机制仍不清楚。
本研究旨在通过 TMT 标记定量蛋白质组学分析 VEN 患儿皮损中蛋白因子的表达变化。
2019 年 1 月至 2019 年 11 月，安徽医科大学第一附属医院皮肤科、博鳌超级医院共纳入 8 例 VEN 患儿（5 例用于实验，3 例用于验证）和 8 例色素痣正常皮肤组织的健康儿童（5 例用于实验，3 例用于验证）。将 VEN 患儿皮损和皮损旁组织定义为 VEN 组，VENC 组，色素痣患儿正常皮肤组织定义为 C 组。我们进行蛋白质组学分析，筛选这些个体皮损中差异表达的蛋白。我们进一步用 Western blotting 验证验证组中 9 个靶向蛋白的相对表达水平。
根据蛋白质组学结果，共鉴定出 4970 种蛋白质，定量了 4770 种蛋白质。其中，VEN 组与 C 组之间病变的至少 1.3 倍上调或下调的蛋白为 586 种（上调：399 种，下调：187 种）。这些蛋白在多种生物学功能中发挥重要作用，如角质化、表皮细胞分化和中性粒细胞激活，并形成了一个复杂的蛋白-蛋白相互作用网络。在 586 种上调或下调的蛋白中，选择了 9 种进行进一步验证。根据 Western blotting 分析结果，角蛋白 6A（Cytokeratin 6A）、BRAF、Filaggrin、S100A7 和 Desmocollin-3 的相对表达水平在 VEN 患儿中明显上调，可能与皮肤屏障功能障碍、表皮细胞过度生长和分化、炎症和免疫以及氧化磷酸化有关，这些都与 VEN 的发病机制有关。
根据 TMT 定量蛋白质组学和 Western blotting 结果，我们鉴定了 8 种有意义的蛋白，即包裹蛋白、NDUFA4、富组蛋白、角蛋白 6A、BRAF、Filaggrin、S100A7 和桥粒芯糖蛋白-3，它们在 VEN 患儿皮损中上调，可能与 VEN 的发病机制有关。我们的研究结果为确定 VEN 的精确发病机制或治疗靶点提供了新的起点。
