钠离子通道β亚基与心房纤颤、室性心动过速的遗传关联研究
发布时间：2019-03-29 10:51
【摘要】：心血管疾病是全世界致残、致死的主要原因,心律失常是在心血管系统疾病中最常见的病症之一。心律失常是由遗传因素和环境因素共同作用的复杂性疾病,严重影响人类健康并成为国民经济的重大负担。全基因组关联分析(genome wide association studies, GWAS)是揭示复杂遗传性疾病易感基因的一种有效方法,但是当前心血管疾病GWAS的研究结果聚焦于常见变异而且已发现变异仅能解释小部分的遗传发病风险——遗传率缺失。所以我们认为罕见变异可能有更大的效应,或许可以解释遗传率缺失现象。为了阐述钠离子通道亚基p4(基因-SCN4B,蛋白质-Navp4)上的罕见变异与心房纤颤(房颤)和室性心动过速(室速)的关系,我们对477例房颤样本和199例室速样本进行了突变筛选。 第一部分是在房颤样本中,采用高分辨率溶解曲线(high resolution melt analysis technology, HRM),对钠离子通道p亚基SCN4B的所有外显子以及外显子连接区域进行突变筛查。经过测序验证后,在4例房颤样本中,一号外显子区域发现一个非同义改变G8S。变异G8S位于Navβ4的信号肽区域,在进化上保守性较高。 然后我们应用病例-对照关联分析研究策略,在两个独立的房颤群体中,探讨非同义改变G8S与房颤患病风险之间的相关性。群体一,944例房颤患者和981例对照群体,群体二,732例房颤患者和1291例对照群体。统计结果结果显示变异G8S的小等位基因A与房颤的患病风险之间存在显著关联(在群体一中P=0.017,OR=3.66；在群体二中P=0.017,OR=3.66)；合并群体后组成一个更大的群体,房颤1676例和对对照群体2272例,结果显示变异G8S小等位基因A与房颤的患病风险之间存在关联极显著(P=4.98x10-4,OR=3.14)。 第二部分是在199例室速患者中,对基因SCN4B的五个外显子和外显子连接区域进行突变筛选。经过测序验证后,发现两个非同义改变,在4例室速样本中,信号肽区域的G8S,在1例室速样本中,三号外显子区域的A145S。A145S位于Navβ4蛋白的胞外区段,在物种进化上保守性较高。经过生物信息学预测,显示G8S和A145S都是有害变异,可以降低NavP4蛋白的稳定性。然后我们应用病例-对照关联分析研究策略,探讨非同义改变G8S与室速患病风险之间的相关性 在两个独立的室速群体中,变异G8S与室速患病风险之间显著相关(在验证群体一中,299例病人对981例正常人,P=5.58×10-5,OR=9.17；在验证群体二中,270例病人对639例正常人,P=4.47×10-3,OR=4.31；在合并群体中,569例病人对1620例正常人,P=1.20×10-7,OR=6.42)。 利用全细胞膜片钳技术在表达了野生型和突变型NavP4的钠离子通道的HEK293细胞上记录钠电流(sodium current,INa),发现与野生型(WT-SCN4B)钠通道相比,突变型钠通道(G8S-SCN4B和A145S-SCN4B)的电生理特性没有显著变化。但是Western blot实验显示,突变型钠通道β4在全细胞水平和膜蛋白水平的表达量明显降低。此项目首次发现SCN4B基因罕见变异与房颤、室速显著相关,为假说“常见疾病,罕见变异”提供了实验证据。
[Abstract]:Cardiovascular disease is the main cause of the world's maiming and death. Arrhythmia is one of the most common conditions in the cardiovascular system. Arrhythmia is a complex disease that has a common role in genetic and environmental factors, which seriously affects human health and becomes a major burden on the national economy. The whole-genome association studies (GWAS) are an effective method to reveal the susceptibility genes of complex genetic diseases, but the results of the study of the GWAS in the present cardiovascular disease focus on the common variation and have found that the variation can only explain the genetic risk _ genetic rate of the small part. So we think that a rare variation may have a greater effect, perhaps to explain the lack of genetic rate. In order to elucidate the relationship between rare variations in sodium channel subunit p4 (gene-SCN4B, protein-Navp4) and atrial fibrillation (atrial fibrillation) and ventricular tachycardia (ventricular tachycardia), we made a mutation screening of 477 atrial fibrillation samples and 199 chamber-rate samples. The first part is a high resolution fusion analysis technique (HRM) in the atrial fibrillation sample, and the mutation screen is carried out on all exons of the sodium ion channel p-subunit SCN4B and the exon-connecting region. A non-synonymous change of G8 was found in exon 1 in 4 AF samples after sequencing. S. The mutant G8S is located in the signal peptide region of Nav-4 and is more conservative in evolution. High. Then we applied case-control association analysis to study the non-synonymous changes between G8S and the risk of atrial fibrillation in two separate AF groups. Correlations. Group I,944 patients with AF and 981 control groups, group II,732 patients with atrial fibrillation and 1291 The results of the statistical results showed that there was a significant association between the small allele A of the variant G8S and the risk of atrial fibrillation (P = 0.017, OR = 3.66 in the population, P = 0.017, OR = 3.66 in the population), a larger population after the group,1676 in AF and 22 for the control group 22. The results showed that there was a significant correlation between the small allele A of the variant G8S and the risk of atrial fibrillation (P = 4.98x10-4, OR = 3). 14). The second part was in 199 chamber-rate patients with five exons and exon-connecting regions of the gene SCN4B. After sequencing and verification, two non-synonymous changes were found, and in the four cell-rate samples, the G8S of the signal peptide region and the A145S of the signal peptide region in the 1 case-rate sample were located in the extracellular region of the Nav-4 protein and on the evolution of the species. The conservation is high. The bioinformatics prediction shows that both G8S and A145S are harmful variants, which can reduce the NavP4 egg. White stability. Then we applied a case-control association analysis study strategy to explore non-synonymous changes in the risk of G8S and ventricular tachycardia The correlation between the variation of G8S and the risk of ventricular tachycardia was significantly correlated in two independent ventricular tachycardia (in the validation group,299 patients were Among the 981 normal subjects, P = 5.58,10-5, OR = 9.17, in the second group, in the first group, in the first group, in the first group, in the first group, in the first group, in the first group, in the first group, in the first group, in the first group, in the first group, in the first group, in the first group, in the first group, in the first group, in the first group, in the first group, in the first group, in the first group, in the first group, in the first group, in the first group, in the first group, the R = 6.42). Sodium current (INa) was recorded on the HEK293 cells expressing the sodium ion channel of the wild type and the mutant NavP4 using a full cell patch clamp technique, and it was found that the mutant sodium channels (G8S-SCN4B and A145S-SCN4B) were not produced as compared to the wild type (WT-SCN4B) sodium channel. The results of Western blot show that the mutant sodium channel-4 is at the whole cell level and membrane protein water. The expression of SCN4B gene was significantly lower in this project. The rare variation of SCN4B gene was found to be related to atrial fibrillation and ventricular tachycardia for the first time.
【学位授予单位】：华中科技大学
【学位级别】：博士
【学位授予年份】：2015
【分类号】：R541.75
[Abstract]:Cardiovascular disease is the main cause of the world's maiming and death. Arrhythmia is one of the most common conditions in the cardiovascular system. Arrhythmia is a complex disease that has a common role in genetic and environmental factors, which seriously affects human health and becomes a major burden on the national economy. The whole-genome association studies (GWAS) are an effective method to reveal the susceptibility genes of complex genetic diseases, but the results of the study of the GWAS in the present cardiovascular disease focus on the common variation and have found that the variation can only explain the genetic risk _ genetic rate of the small part. So we think that a rare variation may have a greater effect, perhaps to explain the lack of genetic rate. In order to elucidate the relationship between rare variations in sodium channel subunit p4 (gene-SCN4B, protein-Navp4) and atrial fibrillation (atrial fibrillation) and ventricular tachycardia (ventricular tachycardia), we made a mutation screening of 477 atrial fibrillation samples and 199 chamber-rate samples. The first part is a high resolution fusion analysis technique (HRM) in the atrial fibrillation sample, and the mutation screen is carried out on all exons of the sodium ion channel p-subunit SCN4B and the exon-connecting region. A non-synonymous change of G8 was found in exon 1 in 4 AF samples after sequencing. S. The mutant G8S is located in the signal peptide region of Nav-4 and is more conservative in evolution. High. Then we applied case-control association analysis to study the non-synonymous changes between G8S and the risk of atrial fibrillation in two separate AF groups. Correlations. Group I,944 patients with AF and 981 control groups, group II,732 patients with atrial fibrillation and 1291 The results of the statistical results showed that there was a significant association between the small allele A of the variant G8S and the risk of atrial fibrillation (P = 0.017, OR = 3.66 in the population, P = 0.017, OR = 3.66 in the population), a larger population after the group,1676 in AF and 22 for the control group 22. The results showed that there was a significant correlation between the small allele A of the variant G8S and the risk of atrial fibrillation (P = 4.98x10-4, OR = 3). 14). The second part was in 199 chamber-rate patients with five exons and exon-connecting regions of the gene SCN4B. After sequencing and verification, two non-synonymous changes were found, and in the four cell-rate samples, the G8S of the signal peptide region and the A145S of the signal peptide region in the 1 case-rate sample were located in the extracellular region of the Nav-4 protein and on the evolution of the species. The conservation is high. The bioinformatics prediction shows that both G8S and A145S are harmful variants, which can reduce the NavP4 egg. White stability. Then we applied a case-control association analysis study strategy to explore non-synonymous changes in the risk of G8S and ventricular tachycardia The correlation between the variation of G8S and the risk of ventricular tachycardia was significantly correlated in two independent ventricular tachycardia (in the validation group,299 patients were Among the 981 normal subjects, P = 5.58,10-5, OR = 9.17, in the second group, in the first group, in the first group, in the first group, in the first group, in the first group, in the first group, in the first group, in the first group, in the first group, in the first group, in the first group, in the first group, in the first group, in the first group, in the first group, in the first group, in the first group, in the first group, in the first group, in the first group, in the first group, in the first group, the R = 6.42). Sodium current (INa) was recorded on the HEK293 cells expressing the sodium ion channel of the wild type and the mutant NavP4 using a full cell patch clamp technique, and it was found that the mutant sodium channels (G8S-SCN4B and A145S-SCN4B) were not produced as compared to the wild type (WT-SCN4B) sodium channel. The results of Western blot show that the mutant sodium channel-4 is at the whole cell level and membrane protein water. The expression of SCN4B gene was significantly lower in this project. The rare variation of SCN4B gene was found to be related to atrial fibrillation and ventricular tachycardia for the first time.
【学位授予单位】：华中科技大学
【学位级别】：博士
【学位授予年份】：2015
【分类号】：R541.75
【相似文献】
相关期刊论文 前10条
1 李宝珠;高炳淼;吴勇;于津鹏;吴潇洒;长孙东亭;罗素兰;;钠离子通道研究进展[J];生物技术;2012年03期
2 张丽敏;康熙雄;丁成峗;;钠离子通道基因多态性在癫痫中的研究进展[J];中华临床医师杂志(电子版);2012年22期
3 王红艳;勾萌;肖蓉;李庆伟;;钠离子通道疾病及其抑制剂生物学功能研究进展[J];生物工程学报;2014年06期
4 唐俊,庄心良,李士通,焦志华,颜涛;咪唑安定临床治疗浓度对海马神经元钠离子通道的作用[J];中国临床药理学与治疗学;2003年05期
5 于耀清,陈军;电压门控性钾、钙、钠离子通道的结构及分类[J];中华神经医学杂志;2005年05期
6 周松;李世根;刘永刚;刘希琴;;钠离子通道及其作用药物研究进展[J];医药导报;2008年07期
7 李耐三;孙桥;;用配基结合方法对电压敏感的钠离子通道的研究[J];南药译丛;1986年02期
8 刘敏,张守兰,林福生,汤凌;河豚毒素与钠离子通道的相互作用[J];卫生毒理学杂志;1999年03期
9 苏敬敬;董强;;丝氨酸蛋白酶调控酸敏感离子通道及上皮钠离子通道机制研究进展[J];神经损伤与功能重建;2009年02期
10 杨雄里;;光感受机制研究的重要突破[J];生理科学进展;1987年01期
相关会议论文 前5条
1 王丽华;张信信;，
本文编号：2449456
本文链接：https://www.wllwen.com/yixuelunwen/xxg/2449456.html
最近更新
教材专著
