本文選題：青少年特發(fā)性脊柱側(cè)凸 + 遺傳學(xué)�。� 參考：《北京協(xié)和醫(yī)學(xué)院》2015年博士論文

【摘要】：研究背景青少年特發(fā)性脊柱側(cè)凸(Adolescent Idiopathic Scoliosis, AIS)是所有脊柱側(cè)凸患者中最常見的一類,不同人群發(fā)病率為0.47-5.2%。在其病因?qū)W研究方面,人們很早就發(fā)現(xiàn)特發(fā)性測凸患者的一級親屬患病風(fēng)險明顯增高,但經(jīng)過多年研究,目前尚未明確提出一個致病基因。近年來,國內(nèi)、國際上有多個研究團(tuán)隊從單核苷酸多態(tài)性(Single Nucleotide Polymorphism, SNP)角度著手進(jìn)行遺傳學(xué)病因研究,共涉及39個AIS相關(guān)候選基因,包括IGF-1、ESR1、LBX-1、 GPR126等。但既往關(guān)于AIS的SNP研究存在諸多不足：各個研究中心的樣本數(shù)量參差不齊,一些小的研究團(tuán)隊僅有數(shù)十名患者,導(dǎo)致研究結(jié)論可信性低；不同人種之間的結(jié)果重復(fù)性差,體現(xiàn)了不同種族之間的遺傳學(xué)背景差異；缺乏大規(guī)模中國北方漢族人群研究資料,不能直接應(yīng)用國外研究的結(jié)果；缺乏深入的功能試驗(yàn),使研究成果停留在“關(guān)聯(lián)性”而不能建立病因?qū)W上的因果關(guān)系。因此,有必要基于我國人群進(jìn)行既往研究的驗(yàn)證工作并對重要候選基因進(jìn)行深入功能試驗(yàn)。研究目的1.建立中國北方漢族人群青少年特發(fā)性脊柱側(cè)凸遺傳學(xué)研究隊列；2.在本隊列中驗(yàn)證既往報道中與AIS發(fā)病相關(guān)的SNP位點(diǎn)；3.針對重點(diǎn)候選基因,通過深入功能分析,嘗試解釋其致病機(jī)制。研究方法確立入組及排除標(biāo)準(zhǔn),收集于北京協(xié)和醫(yī)院骨科行手術(shù)治療的AIS患者外周血樣本及臨床資料,建立研究隊列。通過檢索、回顧文獻(xiàn),確定需要驗(yàn)證的AIS相關(guān)基因及SNP位點(diǎn)。根據(jù)病例-對照研究策略,使用Sequenom MassArray分型平臺,對患者及正常對照的候選SNP進(jìn)行基因分型,并應(yīng)用PLINK及SPSS軟件進(jìn)行統(tǒng)計分析。針對重點(diǎn)基因LBX1,應(yīng)用候選基因關(guān)聯(lián)分析法選取基因及其周圍的SNP位點(diǎn),通過Sequenom MassArray技術(shù)進(jìn)行分型,得到功能試驗(yàn)候選位點(diǎn)。針對該位點(diǎn),使用雙熒光報告素酶系統(tǒng)研究其對LBX1表達(dá)的影響,最終給出致病機(jī)理模型。研究結(jié)果1.本研究共納入中國北方漢族AIS患者180例,性別、民族匹配的正常對照182例；2.通過文件回顧,共納入rs1065755等20處AIS相關(guān)SNP位點(diǎn)；3. rs11190870、rsl2885713和rs2300500三個位點(diǎn)在等位基因關(guān)聯(lián)分析、基因型關(guān)聯(lián)分析以及遺傳模型分析中均與AIS發(fā)病相關(guān),而rs6570507、 rs4753426僅在遺傳模型分析中與AIS具有相關(guān)性,分別符合隱形模型及趨勢模型；4.對LBX1基因應(yīng)用候選基因多態(tài)性分析方法,得到3個與AIS發(fā)病相關(guān)SNP位點(diǎn)：rs11190870,rs1322331及rs625039。其中rs1322331的可信性最高、可進(jìn)行功能試驗(yàn)；5.通過雙熒光報告素酶系統(tǒng)檢驗(yàn),在體外條件下,rs1322331位點(diǎn)為GG純合型時對下游基因的表達(dá)起顯著增強(qiáng)作用,而TT純合型則對下游基因的表達(dá)無顯著作用。結(jié)論1.中國北方漢族人群中,既往文獻(xiàn)報道的rs11190870、rsl2885713.rs2300500、 rs6570507和rs4753426等位點(diǎn)與AIS發(fā)病具有相關(guān)性；2.中國北方漢族人群中,LBXl基因處與AIS發(fā)病相關(guān)聯(lián)且可信度最高的SNP位點(diǎn)是rs1322331；3.GG基因型的rs1322331可上調(diào)其所在的啟動子區(qū)域功能,使下游基因表達(dá)上調(diào)。據(jù)此,我們提出LBX1過量表達(dá)導(dǎo)致AIS發(fā)病的假說。
[Abstract]:Background adolescent idiopathic scoliosis (Adolescent Idiopathic Scoliosis, AIS) is the most common category in all scoliosis patients. The incidence of different populations is 0.47-5.2%. in its etiology research. People have found a significant increase in the risk of first-degree disease in patients with idiopathic protruding, but after years of study, the number of patients with idiopathic scoliosis was found to be significantly higher. In recent years, a number of research teams in the world have conducted genetic studies on Single Nucleotide Polymorphism (SNP), involving 39 AIS related candidate genes, including IGF-1, ESR1, LBX-1, GPR126 and so on. However, there are many previous SNP studies on AIS. There are many shortcomings: the sample size of each research center is uneven, and some small research teams only have dozens of patients, which leads to low credibility of the research conclusions, poor reproducibility of the results between different races, reflecting the genetic background difference between different races, and the lack of a large scale in the northern Han population. With the results of foreign studies, the lack of in-depth functional tests makes the research results stay in "relevance" and can not establish causality in the etiology. Therefore, it is necessary to verify the previous research and test the important candidate genes based on the population of our country. Objective 1. to establish the Han population in Northern China. A cohort of adolescent idiopathic scoliosis genetic studies; 2. in this cohort, the SNP loci associated with AIS were verified in this cohort; 3. for key candidate genes, the pathogenesis was explained by in-depth functional analysis. The research methods established the group and exclusion criteria and collected in the surgical treatment of the Department of orthopedics in Peking Union Medical College Hospital, AIS The patient's peripheral blood samples and clinical data were set up. The AIS related genes and SNP loci were identified by retrieving and reviewing the literature. According to the case-control study strategy, the Sequenom MassArray typing platform was used to classify the patients and the normal control candidate SNP, and the PLINK and SPSS software were used for statistical analysis. In view of the key gene LBX1, the candidate gene association analysis method was used to select the gene and its surrounding SNP loci, and the functional test candidate loci were obtained by Sequenom MassArray technique. In view of the loci, the effect of the double fluorescent reporter system on the expression of LBX1 was studied, and the pathogenic mechanism model was finally given. 1. This study included 180 AIS patients in the Han nationality in northern China, 182 cases of normal control of gender and national match; 2. through document review, a total of 20 AIS related SNP loci were included and three loci of 3. rs11190870, rsl2885713 and rs2300500 were associated with allele correlation analysis, genotype association analysis and genetic model analysis with AIS hair. The disease was related, and rs6570507, rs4753426 only correlated with AIS in the genetic model analysis, which conformed to the stealth model and the trend model respectively. 4. the candidate gene polymorphism analysis method of the LBX1 gene was used to obtain the 3 SNP loci associated with the pathogenesis of AIS: rs11190870, rs1322331 and rs625039., the credibility of rs1322331 was the highest, and the work could be done. Can test; 5. through the double fluorescent reporter system test, under the condition of in vitro, the rs1322331 site is GG homozygous, the downstream gene expression is significantly enhanced, while the TT homozygous type has no significant effect on the downstream gene expression. Conclusion 1. in the Han population of northern China, rs11190870, rsl2885713.rs2300500, R reported in the literature. The loci of s6570507 and rs4753426 are related to the pathogenesis of AIS; 2. in the Han population of northern China, the SNP locus associated with the pathogenesis of AIS and the highest reliability of AIS is rs1322331; the rs1322331 of the 3.GG genotype can increase the function of its promoter region and up-regulation the expression of the downstream genes. Accordingly, we propose that LBX1 overexpression. The hypothesis that causes the onset of AIS.

【學(xué)位授予單位】：北京協(xié)和醫(yī)學(xué)院
【學(xué)位級別】：博士
【學(xué)位授予年份】：2015
【分類號】：R682.3
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本文編號：1825488