全外显子组测序法鉴定两个近亲婚配的低血磷性骨软化家系致病基因突变及功能研究

项目名称： 全外显子组测序法鉴定两个近亲婚配的低血磷性骨软化家系致病基因突变及功能研究

项目编号： No.81200646

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 医学二处

项目作者： 顾洁梅

作者单位： 上海交通大学

项目金额： 23万元

中文摘要： 低血磷性佝偻病/骨软化是以骨痛、肌无力为特征，严重者出现假骨折、骨折甚至骨骼畸形。生化特点为血磷降低，碱性磷酸酶显著升高等。近几年对遗传性低磷性佝偻病/骨软化致病基因研究获得重大进展，相继鉴定了XLH，FGF23，SLC34A3，DMP1和ENPP1等致病基因。但仍有很多遗传性及散发病例的致病基因仍未探明。我们临床诊断了两个近亲婚配低磷骨软化家系，基因筛查中并未发现上述致病基因突变。为此，本项目拟使用全外显子组测序法对2个先证者及其父母进行检测以鉴定致病基因突变；其次开展功能研究，证实其发病机制，将突变体转入体外培养的C3H10T1/2细胞，研究该基因在转染细胞中的表达，并分析该基因转染对细胞生物学行为的影响；最后对42例散发性低磷性佝偻病/骨软化患者检测本研究中找到的突变基因。本项目有望鉴定到导致低磷佝偻病/骨软化的新致病基因，对深化该疾病的分子机制具有重要意义。

中文关键词： 低磷佝偻病/骨软化；全外显子组测序；目标区域；近亲婚配；

英文摘要： Hypophosphatemic rickets/osteomalacia is characterized by bone pain, mucle weakness, and serious patients may suffer from pseudo fractures, fractures and bone deformities. Blood tests show low serum phosphorus and high serum alkaline phosphatase. Recently, significant progress has been made in virulence gene research of this disease. The gene responsible for XLH was identified as PHEX in 1995. After that, The genes mutated in ADHR、HHRH、ARHR1 and ARHR2 have been identi?ed as FGF23, SLC34A3, DMP1 and ENPP1. However, the virulence gene of many sporadic cases have not been found. We make clinical diagnosis of two probands of hypophosphatemic osteomalacia who had family histories of consanguineous marriage. And we failed to find mutations of the genes above in the gene screening. To this end, the project intends to use the whole-exome sequencing method to detect the two probands and their parents to identify the virulence gene mutations. And we will do functional studies after that. The gene mutant will be transferred to the cultured C3H10T1/2 cells. Then we will study the gene expression in transfected cells, and analyze the impact of the gene transfection on the biological behavior of the cells. Finally, we will do direct sequencing to check the identified mutations among 42 sporadic patients.The project is expecte

英文关键词： hypophosphatemic rickets / osteomalacia；whole exome sequencing；target area；consanguineous marriage；