整合分析证实骨保护素基因与骨质疏松症的相关性

doi:10.24920/003466

摘要/Abstract

摘要： 目的通过整合分析的方法证实骨保护素（osteoprotegerin,OPG）基因及其遗传变异与骨质疏松症（osteoporosis,OP）的相关性。方法我们使用KGG软件进行基于基因的关联分析,整合了所有可公开获取的基于单核苷酸多态性（single-nucleotide polymorphism,SNP）的P值,并获得OPG基因的总体P值。筛选显著的SNP表达数量性状位点（expression quantitative trait locus,eQTL）。采用Meta分析的方法整合文献报道中OPG变异基因与骨密度（bone mineral density,BMD)的关联性。使用双荧光素酶报告基因系统对有关联性的基因进行体外功能验证。结果在基于基因的关联分析中, OPG基金的整体P值在股骨颈（femoral neck,FN）骨密度（bone mineral density, BMD）为6.24×10 ^-13,在腰椎（lumbar spine,LS）的BMD为7.37×10 ^-17,表明OPG基因对OP的重要性。公开的eQTL数据库确定了5个eQTL,它们在FN和LS部位对OPG基因存在顺式调控效应。文献检索发现rs2073617（又称为T950C）是热点SNP。除了GEFOS-2的研究外,在PubMed检索到13项关于rs2073617的相关研究。通过荟萃分析揭示了FN（P=0.047）和LS（P=0.025）的BMD在TT,TC和CC基因型之间均存在显著差异,证明了T950C多态性与BMD之间的关联。在等位基因C存在的293T细胞中,荧光素酶基因表达显著高于等位基因T（t=-9.47,P<0.01）。结论整合分析进一步证实了OPG基因对OP的重要性,以及T950C多态性与OP的骨密度的相关性。该策略可为其它疾病相关基因的功能解释提供参考。

关键词: 骨保护素, 骨质疏松症, 单核苷酸多态性, 整合分析

Abstract: Objective This study aimed to verify the association between osteoprotegerin gene (OPG) and its variants with osteoporosis (OP) by performing integrative analysis.Methods We used the KGG software to perform gene-based association analysis, which integrated all publicly available single-nucleotide polymorphism (SNP)-based P values and obtained an overall P value for the OPG. The significant SNPs were screened for expression quantitative trait loci (eQTLs). Meta-analysis was used to combine the associations between the variants of OPG and bone mineral density (BMD) reported in the literatures. Then we performed dual-luciferase reporter gene systems for the functional verification of the variants of OPG in vitro.Results In the gene-based association analysis, the over all P value of OPG was 6.24×10 ^-13for BMD at femoral neck (FN) and 7.37×10 ^-17 for BMD at lumbar spine (LS), indicating the importance of OPG for OP. The publicly available eQTL database identified 5 eQTLs which exert cis-regulation effects on OPG at FN and LS. Literature searching found that rs2073617 (known as T950C) was the hot spot SNP. There were 13 relevant studies on rs2073617 besides the GEFOS-2 study identified from the PubMed. Significant differences among TT, TC and CC genotypes at FN (P= 0.047) and LS (P= 0.025) were shown by meta-analysis, demonstrating the associations between T950C polymorphism and BMD. Luciferase gene expression was significantly higher at the presence of allele C than allele T in the 293T cells (t=-9.47, P<0.01). Conclusion The integrative analysis further confirmed the importance of OPG in OP and the correlation of T950C polymorphism with BMD of OP. The strategy can be used as a reference for functional interpretation of other disease-related genes.

Key words: osteoprotegerin, osteoporosis, single-nucleotide polymorphism, integrative analysis

基金资助: 国家自然科学基金((No. 31271336).)

Tang Hui,Zhu Xiaowei,Wu Longfei,Mo Xingbo,Deng Feiyan,Lei Shufeng. Integrative Analysis Confirmed the Association between Osteoprotegerin and Osteoporosis[J].Chinese Medical Sciences Journal, 2019, 34(2): 147-156.

图/表 5

参考文献 34

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SNP	Position	Score^*	FN P value^#	LS P value^#	Number of studies^?
rs6469789	chr8:120029842..120029842	0.01	4.89×10^-14	1.12×10^-17	0
rs3134063	chr8:120028838..120028838	0.06	3.13×10^-9	8.06×10^-15	1
rs3134058	chr8:120023289..120023289	0.03	1.25×10^-3	4.09×10^-9	0
rs1385503	chr8:120031667..120031667	0.07	1.05×10^-3	4.37×10^-9	0
rs2073617	chr8:120033464..120033464	0.46	2.50×10^-9	6.19×10^-15	13

Study author	Publication year	Country	Study design	Population	Age (year)	Sex	Producer of BMD measurement	Anatomic sites measured
Langdahl BL, et al.^[16]	2002	Denmark	case control	268 osteoporosis patients; 327 normal people	52-72	male/ female	Hologic QDR 1000 or Norland Corp.	LS; FN; TH
Yamada Y, et al.^[17]	2003	Japan	cross- sectional	1095 women	40-79	female	Hologic QDR 4500	TB; LS; FN; TR; WT
Cheng Q, et al.^[18]	2004	China	cross- sectional	259 postmenopausal women	55-71	female	Norland XR -36	LS; FN; TR; WT
Vidal C, et al.^[19]	2006	Britain	cross- sectional	126 postmenopausal women	40-75	female	Norland Corp.	LS; FN; TR; WT
Wynne F, et al.^[20]	2002	Malta	cross- sectional	381 postmenopausal women; 130 premenopausal women	40-70	female	Lunar DPX1Q	LS; FN
Ueland T, et al.^[21]	2007	Australia	cross- sectional	1469 postmenopausal women	75±3	female	Hologic Corp.	FN; TH
Wang C, et al.^[22]	2009	China	cross- sectional	80 postmenopausal women with osteoporosis	>50	female	Hologic QDR 2000	LS; FN; TR; IN; TH

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