发育性髋关节脱位家系全基因组外显子测序研究  

Whole-exome analysis in pedigree of developmental dysplasia of the hip

作　　者：王隼[1] 焦勤[1] 周在威[1] 吴静[2] 范玲燕[1] 陈梦婕[1] 王一臣[1] 马峰[1] 应灏[1] 赵利华[1] Wang Sun;Jiao Qin;Zhou Zaiwei;Wu J ing;Fan Lingyan;Chen Mengjie;Wang Yichen;Ma Feng;Ying Hao;Zhao Lihua(Department of Orthopedics, Affiliated Shanghai Children＇s Hospital, Shanghai J iao Tong University, Shanghai 200062, China （Wang S, Jiao Q, Zhou ZW, Wu J, Fan LY, Chen MJ , Wang YC, Ma F, Ying H, Zhao LH;Institute of Pediatric Translational Medicine, Shanghai Children＇s Medical Center, School of Medicine, Shanghai Jiao Tong University, Shanghai 200127,China （Wu J)

机构地区：[1]上海市儿童医院/上海交通大学附属上海市儿童医院骨科,200062 [2]上海交通大学医学院附属上海儿童医学中心儿科转化医学研究所

出　　处：《中华小儿外科杂志》2018年第3期171-177,共7页Chinese Journal of Pediatric Surgery

基　　金：国家自然科学基金（81401763）;上海市卫生计生委科研项目（20144Y0176）;上海市科学技术委员会科研项目（12DZ2295006）;上海交通大学医学院“985”计划基金（YBKL2013008）

摘　　要：目的对一个三代四人发病的DDH家系进行研究，以期发现与DDH发病相关的致病基因或易感基因。方法对一个三代四人发病的DDH家系中的3个DDH患者和2个健康成员的血液样本进行全外显子测序并对测序结果进行生物信息学分析。采用Sanger测序验证分析获得的致病候选变异在其他可获得血液样本的家系成员中突变情况。结果外显子测序结果经过生物信息学分析后发现有2个基因可能与DDH的发病有关：①骨形成蛋白2诱导激酶基因（bone morphogenic proteins-2-inducible kinase gene，BMP2K）中存在的改变蛋白修改功能的多核苷酸缺失/插入突变（c.1432_1440delCAGCAGCAG, p．Gln478_480del或c.1440_1441insCAG, p．Gln480ins），该突变位于第4号染色体第11外显子。②前列腺素F2α受体基因（prostaglandin F2α receptor，PTGFR）中存在的终止密码子突变（c.C922T, p．R308X），该突变位于第1号染色体PTGFR基因的第2/4外显子。DDH患者和具有潜在突变携带可能的正常母亲血液样本中均有BMP2K和PTGFR突变，而作为对照的正常姐姐的血液样本中没有BMP2K和PTGFR突变。Sanger测序验证结果显示，家系中的全部4例DDH患者（100%）和有血缘关系的12例正常家系成员中的3例（25%）存在BMP2K基因的杂合突变和PTGFR的终止密码子突变，而8例无血缘关系的正常家系成员无BMP2K基因和PTGFR基因突变。结论研究结果提示BMP2K基因的杂合突变和PTGFR的终止密码子突变可能是导致该家系DDH发病的致病基因或易感基因。结合家系的宝贵遗传信息和二代测序的高效、高通量技术是发现与DDH发病相关的致病位点的有效途径。ObjectiveDevelopmental dysplasia of the hip （DDH） is a complex disorder of hip joint affecting 1‰-5‰ of newborns.Despite unknown genetic impact on DDH, no functionally comparable genetic event has been found.Here we reported the variants contributing to DDH susceptibility in a family with four individuals affected across three generations.MethodsWhole-exome sequencing was performed for three affected and two unaffected individuals of a DDH pedigree.Potential candidate variants were confirmed by Sanger sequencing and then validated in available family members.ResultsNot previously reported mutations were identified in two genes.One was protein-altering heterozygous, deletion or insertion mutation （c.1432_1440delCAGCAGCAG, p.Gln478_480del or c. 1440_1441insCAG, p.Gln480ins） in exon 11 of chromosome 4 in bone morphogenic proteins-2-inducible kinase （BMP2K） and another one termination codon mutation in exon 2/4 of chromosome 1 in prostaglandin F2α receptor （PTGFR）. Both mutations in BMP2K and PTGFR were shared by DDH-affected members and unaffected grandmother deemed to be a carrier of potential mutations but not by unaffected normal control aunt.Mutations were present in 4/4 （100%） of affected family members, 3/12 （25%） of unaffected related family members and 0/8 （0%） of unaffected unrelated family members by Sanger sequencing.ConclusionsBMP2K and PTGFR variants are probably DDH susceptibility inducing candidates.And a combination of pedigree information and next generation sequencing is effective for identifying pathogenic sites associated with DDH.

关 键 词：髋关节脱位 外显子 序列测定 基因突变 易感基因 

分 类 号：R726.8[医药卫生—儿科]