鈉離子通道β亞基與心房纖顫、室性心動過速的遺傳關(guān)聯(lián)研究
發(fā)布時間：2019-03-29 10:51
【摘要】：心血管疾病是全世界致殘、致死的主要原因,心律失常是在心血管系統(tǒng)疾病中最常見的病癥之一。心律失常是由遺傳因素和環(huán)境因素共同作用的復(fù)雜性疾病,嚴(yán)重影響人類健康并成為國民經(jīng)濟的重大負(fù)擔(dān)。全基因組關(guān)聯(lián)分析(genome wide association studies, GWAS)是揭示復(fù)雜遺傳性疾病易感基因的一種有效方法,但是當(dāng)前心血管疾病GWAS的研究結(jié)果聚焦于常見變異而且已發(fā)現(xiàn)變異僅能解釋小部分的遺傳發(fā)病風(fēng)險——遺傳率缺失。所以我們認(rèn)為罕見變異可能有更大的效應(yīng),或許可以解釋遺傳率缺失現(xiàn)象。為了闡述鈉離子通道亞基p4(基因-SCN4B,蛋白質(zhì)-Navp4)上的罕見變異與心房纖顫(房顫)和室性心動過速(室速)的關(guān)系,我們對477例房顫樣本和199例室速樣本進(jìn)行了突變篩選。 第一部分是在房顫樣本中,采用高分辨率溶解曲線(high resolution melt analysis technology, HRM),對鈉離子通道p亞基SCN4B的所有外顯子以及外顯子連接區(qū)域進(jìn)行突變篩查。經(jīng)過測序驗證后,在4例房顫樣本中,一號外顯子區(qū)域發(fā)現(xiàn)一個非同義改變G8S。變異G8S位于Navβ4的信號肽區(qū)域,在進(jìn)化上保守性較高。 然后我們應(yīng)用病例-對照關(guān)聯(lián)分析研究策略,在兩個獨立的房顫群體中,探討非同義改變G8S與房顫患病風(fēng)險之間的相關(guān)性。群體一,944例房顫患者和981例對照群體,群體二,732例房顫患者和1291例對照群體。統(tǒng)計結(jié)果結(jié)果顯示變異G8S的小等位基因A與房顫的患病風(fēng)險之間存在顯著關(guān)聯(lián)(在群體一中P=0.017,OR=3.66；在群體二中P=0.017,OR=3.66)；合并群體后組成一個更大的群體,房顫1676例和對對照群體2272例,結(jié)果顯示變異G8S小等位基因A與房顫的患病風(fēng)險之間存在關(guān)聯(lián)極顯著(P=4.98x10-4,OR=3.14)。 第二部分是在199例室速患者中,對基因SCN4B的五個外顯子和外顯子連接區(qū)域進(jìn)行突變篩選。經(jīng)過測序驗證后,發(fā)現(xiàn)兩個非同義改變,在4例室速樣本中,信號肽區(qū)域的G8S,在1例室速樣本中,三號外顯子區(qū)域的A145S。A145S位于Navβ4蛋白的胞外區(qū)段,在物種進(jìn)化上保守性較高。經(jīng)過生物信息學(xué)預(yù)測,顯示G8S和A145S都是有害變異,可以降低NavP4蛋白的穩(wěn)定性。然后我們應(yīng)用病例-對照關(guān)聯(lián)分析研究策略,探討非同義改變G8S與室速患病風(fēng)險之間的相關(guān)性 在兩個獨立的室速群體中,變異G8S與室速患病風(fēng)險之間顯著相關(guān)(在驗證群體一中,299例病人對981例正常人,P=5.58×10-5,OR=9.17；在驗證群體二中,270例病人對639例正常人,P=4.47×10-3,OR=4.31；在合并群體中,569例病人對1620例正常人,P=1.20×10-7,OR=6.42)。 利用全細(xì)胞膜片鉗技術(shù)在表達(dá)了野生型和突變型NavP4的鈉離子通道的HEK293細(xì)胞上記錄鈉電流(sodium current,INa),發(fā)現(xiàn)與野生型(WT-SCN4B)鈉通道相比,突變型鈉通道(G8S-SCN4B和A145S-SCN4B)的電生理特性沒有顯著變化。但是Western blot實驗顯示,突變型鈉通道β4在全細(xì)胞水平和膜蛋白水平的表達(dá)量明顯降低。此項目首次發(fā)現(xiàn)SCN4B基因罕見變異與房顫、室速顯著相關(guān),為假說“常見疾病,罕見變異”提供了實驗證據(jù)。
[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.
【學(xué)位授予單位】：華中科技大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2015
【分類號】：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.
【學(xué)位授予單位】：華中科技大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2015
【分類號】：R541.75
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