摘要
目的 :临床上发现一组特殊病例,一个家族4代人中均发现有同一种神经肌肉性疾病的患者,症状具体表现为上肢肘关节有不同程度的伸直受限,下肢踝关节似马蹄样畸形,足部高弓畸形、无明显内翻内收现象。本研究拟通过全基因组测序,探讨其发病的致病基因及分子机制。方法:利用高通量测序技术,分别对患儿、具有相似症状的患儿父亲及健康母亲的血液样本进行全基因组测序。利用生物信息学分析筛选疾病相关基因突变,并进一步收集该家族中的其他患病及健康成员的血液样本,利用PCR技术进行验证。结果:生物信息学分析发现一系列突变基因,包括ANXA3基因突变[chr4,c.C820T(p.R274*)]、ATP6V1E2基因突变[chr2,c.G136A(p.V46M)]和HIST1H3A基因突变[chr6,c.C205T(p.Q69*)]等。基于家族更多成员血液样本的PCR实验结果进一步证实,ANXA3为该疾病潜在的致病基因,突变可导致其蛋白质高级结构的改变,可能与该疾病发生有关。结论:本研究为从分子水平上了解该家族遗传性疾病的发生提供了一定的理论参考。
Objective：We found a special case in clinic：4 generations in a family were detected neuromuscular disorders. The symptoms included the upper elbows had different degrees of limited straight,lower limbs showed equines deformity,and feet were talipes cavus and were not detected obvious varus and adduction. This research aimed to study genetic mutation and molecular mechanism in crucial gene using whole-genome sequencing. Methods：Blood samples from sick child,father with the similar symptom and healthy mother were used to perform whole-genome sequencing by high-throughput sequencing technique. Bioinformatic analysis was used to filter gene mutations associated with disease. Further PCR validations were performed using blood samples from patients and healthy relatives in the family. Results：A series of gene mutations were obtained based on bioinformatics,including ANXA3 gene mutation（chr4,c.C820T（p.R274＊））,HIST1H3 A gene mutation（chr 6,c.C205T（p.Q69＊）） and MTHFR gene mutation（c.T784C（p.Y262H））. PCR validation using blood samples from the family indicated that ANXA3 may be potential causing gene,and the mutation led to change of three-dimensional structure of ANXA3 protein. Conclusion：These results implicated that the ANXA3 may be related with the occurrence of the disease. This study provides certain reference for further understanding the genetic disease from the molecular levels.
出处
《南京医科大学学报（自然科学版）》
CAS
CSCD
北大核心
2015年第12期1757-1760,共4页
Journal of Nanjing Medical University(Natural Sciences)
基金
国家自然科学基金资助(61271054)
南京医科大学科技发展基金重点项目(2013NJMU095)
