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Abstract：
Mastocytosis, a rare blood disorder characterized by the proliferation of clonal abnormal mast cells, has a variegated clinical spectrum and diagnosis is often difficult and delayed. Recently we proposed the cathepsin inhibitor cystatin D-R26 as a salivary candidate biomarker of systemic mastocytosis (SM). Its C26 variant is able to form multiprotein complexes (mPCs) and since protein-protein interactions (PPIs) are crucial for studying disease pathogenesis, potential markers, and therapeutic targets, we aimed to define the protein composition of the salivary cystatin D-C26 interactome associated with SM. An exploratory affinity purification-mass spectrometry method was applied on pooled salivary samples from SM patients, SM patient subgroups with and without cutaneous symptoms (SM+C and SM-C), and healthy controls (Ctrls). Interactors specifically detected in Ctrls were found to be implicated in networks associated with cell and tissue homeostasis, innate system, endopeptidase regulation, and antimicrobial protection. Interactors distinctive of SM-C patients participate to PPI networks related to glucose metabolism, protein S-nitrosylation, antibacterial humoral response, and neutrophil degranulation, while interactors specific to SM+C were mainly associated with epithelial and keratinocyte differentiation, cytoskeleton rearrangement, and immune response pathways. Proteins sensitive to redox changes, as well as proteins with immunomodulatory properties and activating mast cells, were identified in patients; many of them were involved directly in cytoskeleton rearrangement, a process crucial for mast cell activation. Although preliminary, these results demonstrate that PPI alterations of the cystatin D-C26 interactome are associated with SM and provide a basis for future investigations based on quantitative proteomic analysis and immune validation.
摘要：
肥大细胞增多症,一种罕见的血液疾病，其特征是克隆异常肥大细胞的增殖，具有多样化的临床频谱，诊断通常很困难且延迟。最近，我们提出了组织蛋白酶抑制剂胱抑素D-R26作为系统性肥大细胞增多症（SM）的唾液候选生物标志物。它的C26变体能够形成多蛋白复合物（mPCs），并且由于蛋白质-蛋白质相互作用（PPI）对于研究疾病发病机理至关重要，潜在标记,和治疗目标，我们旨在确定与SM相关的唾液胱抑素D-C26相互作用组的蛋白质组成。一种探索性亲和纯化-质谱方法应用于SM患者的唾液样本，有和没有皮肤症状的SM患者亚组（SM+C和SM-C），和健康控制（Ctrls）。发现在Ctrls中特别检测到的相互作用者与细胞和组织稳态相关的网络有关。先天系统，内肽酶调节，和抗菌保护。SM-C患者独特的相互作用者参与与葡萄糖代谢相关的PPI网络，蛋白质S-亚硝基化，抗菌体液反应，中性粒细胞脱颗粒,而SM+C特异性的相互作用物主要与上皮和角质形成细胞分化有关，细胞骨架重排，和免疫反应途径。对氧化还原变化敏感的蛋白质，以及具有免疫调节特性和激活肥大细胞的蛋白质，在患者中被发现；他们中的许多人直接参与细胞骨架重排，对肥大细胞活化至关重要的过程。虽然是初步的，这些结果表明胱抑素D-C26相互作用组的PPI改变与SM相关,并为基于定量蛋白质组学分析和免疫验证的未来研究提供了基础.
