骨质疏松易感SNP rs4325274通过增强子远程调控SOX6基因的功能机制研究  

The osteoporosis susceptible SNP rs4325274 remotely regulates the SOX6 gene through enhancers

作　　者：妥晓梅 朱东丽[1,2] 陈晓峰 荣誉[1] 郭燕 杨铁林[1,2] Xiaomei Tuo;Dongli Zhu;Xiaofeng Chen;Yu Rong;Yan Guo;Tielin Yang(Biomedical Informatics&Genomics Center,Key Laboratory of Biomedical Information Engineering of Ministry of Education,School of Life Science and Technology,Xi'an Jiaotong University,Xi'an 710049;Research Institute of Zhejiang Xi'an Jiaotong University,Hangzhou 311215)

机构地区：[1]西安交通大学生命科学与技术学院,生物医学信息工程教育部重点实验室,生物医学信息与基因组学中心,西安710049 [2]浙江西安交通大学研究院,杭州311215

出　　处：《遗传》2020年第9期889-897,共9页Hereditas（Beijing)

基　　金：国家自然科学基金面上项目(编号:31771399,31970569);中国博士后基金项目(编号:2019M650261);陕西省自然科学基础研究计划项目(编号:2020JQ-026);浙江省自然科学基金(编号:GF18C060003)资助。

摘　　要：骨质疏松症是一种典型的多基因复杂疾病,遗传力高达85%,其发病率已跃居常见疾病的第5位。尽管已经鉴定出大量骨质疏松易感SNP,但大多数SNP位点位于基因组非编码区,且功能机制未知。本研究旨在通过生物信息学分析和功能实验探究骨质疏松非编码功能性易感SNP rs4325274的分子调控机制。首先,通过表观注释发现该SNP所在区域处在增强子上,eQTL和Hi-C分析结果发现SNP调控的潜在靶基因是SOX6;然后,利用多种数据库进行Motif预测,并结合GEO数据库中的ChIP-seq数据分析进行了验证,结果发现转录因子HNF1A更倾向于结合SNP rs4325274-G碱基;进一步通过双荧光素酶报告基因实验验证了该SNP对SOX6基因表达的增强作用;最后,利用shRNA敲低转录因子HNF1A实验,检测靶基因SOX6的表达变化。以上研究结果初步解析了非编码区功能性SNPrs4325274作为增强子远程调控SOX6基因表达的分子机制,为复杂疾病非编码易感SNP的遗传调控研究提供新思路。Osteoporosis is a typical polygenic disease,and its heritability is as high as 85%.The incidence of osteoporosis has jumped to the fifth among the common diseases.Although a large number of osteoporosis-susceptible SNPs have been identified,most of them are in the non-coding regions of the genome and the functional mechanisms are unknown.The purpose of this study was to explore the function of non-coding osteoporosis-susceptible SNP rs4325274 and dissect the molecular regulatory mechanisms through integrating bioinformatics analysis and functional experiments.Firstly,we found the SNP rs4325274 resided in a putative enhancer element through functional annotation.eQTL and Hi-C analysis found that the SOX6 gene might be a potential distal target of rs4325274.We conducted the motif prediction using multiple databases and verified the result using ChIP-seq data from GEO database.The result showed that the transcription factor HNF1A could preferentially bind to SNP rs4325274-G allele.We further demonstrated that SNP rs4325274 acted as an enhancer regulating SOX6 gene expression by using dual-luciferase reporter assays.Knockdown of HNF1A decreased the SOX6 gene expression.Taken together,our results uncovered a new mechanism of a non-coding functional SNP rs4325274 as a distal enhancer to modulate SOX6 expression,which provides new insights into deciphering molecular regulatory mechanisms underlying non-coding susceptibility SNPs on complex diseases.

关 键 词：SNP rs4325274 SOX6基因 转录因子 骨质疏松症发生机制 

分 类 号：R580[医药卫生—内分泌]