Crouzon综合征中FGFR2^C342Y基因突变的血清exosome蛋白组学变化

Proteomic analysis of serum exosomes derived from Crouzon syndrome patients carrying a mutant FGFR2^C342Y

目的研究FGFR2C342Y基因突变引起的Crouzon综合征患者与正常人血清来源外泌体的蛋白组分的差异。方法筛选出Crouzon综合征FGFR2^C342Y突变患者的血清作为实验组，健康献血者的血清作为对照组，并使用超速离心法从中提取外泌体，展示了同位素标记相对和绝对定量（iTRAQ）蛋白组学分析结果，利用IPA通路分析软件分析其差异蛋白，以探讨exosome差异蛋白在颅缝早闭中的作用。结果通过质谱分析共检测到62个蛋白，其中22个蛋白与骨相关数据库重合，其功能主要关于发育异常、遗传性异常、肌肉骨骼系统发育和功能、组织形态、视觉系统发育和功能等，均与Crouzon综合征的临床表现相关。网络分析发现“组织损伤和异常、血液系统的发育和功能、细胞信号转导和相互影响”相关的网络。结论我们的研究显示了FGFR2^C342Y综合征患者的蛋白组学特点，展示了差异蛋白参与的功能和网络，并找到了关键蛋白FN1，FN1促进成骨细胞黏附增殖和矿化，这个蛋白可能对于Crouzon综合征的进一步研究提供新思路。

Objective Reveal the global expression profile of serum exosomal proteins of Crouzon syndrome patients. Methods We isolated microvesicles from serum of Crouzon children with a C342Y mutation in FGFR2 by uhracentrifugation, which were further characterized by electron microscopy and immunoblotting. The protein profiling in normal subjects and Crouzon patients was systematically compared by iTRAQ proteomic analysis. Results The result demonstrated that microvesicles were between 30--100 nm in diameter, round shape with cup-like concavity and expressed exosomal marker tumor susceptibility gene （TSG） 101 and flotillin （Flot） 1. We identified a total number of 62 proteins, among which 22 proteins overlap with ExoCarta database and were different between the Crouzon patient and the normal subject. The Ingenuity Pathway Analysis showed that the functions of these proteins are mostly involved in Developmental Disorder, Hereditary Disorder, Skeletal and Muscular Disorders, which are all related to the clinical manifestations of Crouzon syndrome. In addition, the proteins were focused on the network of ＂Organismal Injury and Abnormalities, Hematological System Development and Function, Cell-To-Cell Signaling and Interaction＂. The central protein FN1 was presented as the key protein in the network. Conclusions Our data demonstrated that serum exosomes harbor informative proteins and FN1 was selected as a potential candidate for its role in promoting osteoblast adhesion, proliferation and mineralization.