Fgfr2c(C342Y/+)基因突变导致颅缝早闭症Crouzon综合征的分子机制研究

项目名称： Fgfr2c(C342Y/+)基因突变导致颅缝早闭症Crouzon综合征的分子机制研究

项目编号： No.81201483

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 医学四处

项目作者： 杨娴娴

作者单位： 上海交通大学

项目金额： 23万元

中文摘要： 颅缝早闭症Crouzon综合征是常见的先天性颅颌面畸形之一，由FFGR2突变所致，确切分子机制不清。Fgfr2c（C342Y/+）小鼠为针对该突变的动物模型，尚未有利用其开展基因分子调控机制的研究报道。课题组前期在患者颅缝组织基因芯片研究中发现RBP4、GPC3、C1QTNF3等新致病基因，并在Fgfr2c（C342Y/+）小鼠颅缝组织中验证，结果提示新基因可能处于FGFR2下游通路。申请者设想：上述新基因是否通过FGFR2信号通路调控颅缝细胞增殖与分化失衡，从而导致颅缝早闭？基于此，本课题首先研究新基因在小鼠颅缝细胞中的表达定位；通过激活、抑制FGFR2信号通路，探讨FGFR2调控新基因的作用机制；通过新基因过表达及沉默，双向检测主要通路蛋白的表达改变，并验证新基因与通路蛋白的相互作用。旨在初步阐明新基因通过FGFR2信号通路的作用模式，为进一步开展针对新致病基因的靶向治疗提供理论依据。

中文关键词： 颅缝早闭症；Crouzon综合征；Fgfr2c(C342Y/+)基因突变；FGF2信号通路；分子机制

英文摘要： Crouzon syndrome is one of the frequent congenital deformities with patients exhibit premature fusion of multiple cranial sutures resulting in craniosynostosis during early childhood. The C342Y gain-of-function substitution in the Fgfr2 gene is a commonly reported mutation for this condition. A previous microarray study in human identified a number of novel genes incluing RBP4, GPC3 and C1QTNF3 were downregulated during this process and were similarly expressed in the Fgfr2c(C342Y/+) Crouzon mouse model. This strongly suggest they are downstream effector of FGF2 signaling. In this study, the onset of cranial suture fusion in Fgfr2c(C342Y/+) mice will be assessed from E16.5 to P10 using microCT and histology. Expression levels and localization of target genes with osteogenic markers of coronal suture-derived cell cultures from wildtype and Fgfr2c(C342Y/+) mice during osteogenesis will be determined using Realtime PCR and immunocytochemistry respectively. The metabolic and proliferation rates of coronal suture-derived and parietal bone-derived cell cultures will also be assessed by MTS and DNA assay. We will further indentify levels of target gene expression by both FGF2 stimulation and FGFR2 inhibition in these cells. Levels of FGF2 signaling proteins expression will then be detacted after overexpression and siR

英文关键词： craniosynostosis；Crouzon syndrome；Fgfr2c(C342Y/+) gene mutation；FGF2 signaling；molecular mechanism