本文選題：先天性心臟病　切入點(diǎn)：染色體芯片分析　出處：《上海交通大學(xué)》2014年博士論文

【摘要】：目的:本研究旨在評(píng)價(jià)染色體芯片分析(CMA)對(duì)先天性心臟病(CHD)的分子診斷率,比較綜合征型CHD與單純型CHD之間拷貝數(shù)變異(CNV)檢出率的差異;分析CHD臨床表型與CNV的相關(guān)性;并利用正常對(duì)照數(shù)據(jù)及基因優(yōu)選分析鑒定新的CHD候選相關(guān)位點(diǎn)及基因。方法:本研究系統(tǒng)回顧了在2006年12月至2013年4月期間就診于波士頓兒童醫(yī)院(BCH)心臟科的CHD患兒的詳細(xì)臨床資料及臨床CMA結(jié)果。所有納入研究的BCH CHD病例被分為單純型CHD和綜合征型CHD兩類,并依據(jù)患兒的表型進(jìn)一步將該隊(duì)列的CHD病例劃分成9個(gè)亞型。本研究同時(shí)還將在2011年9月至2012年3月期間就診于上海兒童醫(yī)學(xué)中心(SCMC)心臟外科的圓錐動(dòng)脈干畸形患兒作為研究對(duì)象。BCH及SCMC CHD隊(duì)列的CMA分析分別基于Agilent 4×180K SNP+CGH芯片和Affymetrix Cytoscan HD芯片平臺(tái)。所有經(jīng)確定的非多態(tài)性CNV依據(jù)美國(guó)醫(yī)學(xué)遺傳協(xié)會(huì)(ACMG)指南進(jìn)行臨床致病性的評(píng)估。統(tǒng)計(jì)BCH隊(duì)列中CMA分析對(duì)單純型CHD和綜合征型CHD的分子診斷率,計(jì)算并比較該隊(duì)列中各個(gè)CHD亞型間CNV頻率的差異。綜合兩個(gè)隊(duì)列的CMA分析結(jié)果,通過(guò)與正常對(duì)照組的比對(duì),篩選新的CHD候選相關(guān)CNV。最后,結(jié)合在線分析工具、基因表達(dá)數(shù)據(jù)庫(kù)及通路分析,設(shè)計(jì)了一套基因優(yōu)選分析流程以鑒定新的CHD候選相關(guān)基因。結(jié)果:納入本研究的CHD病例共包括514例CHD患兒,其中422例來(lái)自BCH隊(duì)列,92例來(lái)自SCMC隊(duì)列。對(duì)于BCH隊(duì)列,臨床CMA檢測(cè)對(duì)CHD的整體分子診斷率為12.8-18.48%。排除12例攜帶染色體非整倍體的病例,CMA對(duì)綜合征型CHD的分子診斷率為14.11-20.56%,而對(duì)單純型CHD的診斷率為4.32-9.26%。在所有CHD亞型中,致病性CNV在圓錐動(dòng)脈干畸形及心臟間隔缺損病例中的頻率最高。SCMC隊(duì)列的分子診斷率略低(8.7-12%)。此外,本研究篩選出4個(gè)重現(xiàn)次數(shù)≥3且在CHD隊(duì)列中顯著富集的染色體區(qū)域(4q末端區(qū)域、15q11.2、16p12.2及Yp11.2)作為新的CHD候選相關(guān)性位點(diǎn)。最后通過(guò)基因優(yōu)選流程,鑒定出20個(gè)新的CHD候選相關(guān)基因。結(jié)論:本研究的CNV分析結(jié)果為臨床CMA平臺(tái)作為診斷CHD的一線實(shí)驗(yàn)檢測(cè)方法提供了重要證據(jù)。盡管綜合征型CHD的致病性CNV檢出率更高,但CMA對(duì)單純型CHD仍具有一定的診斷價(jià)值,尤其對(duì)于患圓錐動(dòng)脈干畸形及心臟間隔缺損的病例CNV的檢出率更高。致病性CNV的分析及篩選為候選基因的鑒定提供了豐富的資源。
[Abstract]:Objective: to evaluate the molecular diagnostic rate of chromosomal microarray analysis (CMA) for congenital heart disease (CHD), to compare the difference in the detection rate of CHD between syndromic CHD and simplex CHD, and to analyze the correlation between CHD clinical phenotype and CNV. The new candidate loci and genes of CHD were identified by normal control data and gene analysis. Methods: the CHD from December 2006 to April 2013 in cardiac department of Boston Children's Hospital was systematically reviewed. Detailed clinical data and clinical CMA results of children. All BCH CHD cases included in the study were divided into two groups: simple CHD and syndromic CHD. The cohort of CHD cases were further divided into 9 subtypes according to their phenotypes. From September 2011 to March 2012, we will also visit Shanghai Children's Medical Center (SCC) from September 2011 to March 2012 to study conical artery trunk malformation in cardiac surgery. The CMA analysis of SCMC CHD cohort and. BCH cohort were based on Agilent 4 脳 180K SNP CGH and Affymetrix Cytoscan HD chip, respectively. All identified non-polymorphic CNV were evaluated for clinical pathogenicity according to the American Medical genetic Association (ACMG) guidelines. To estimate the molecular diagnostic rate of CMA analysis in BCH cohort for simplex CHD and syndromic CHD, The differences of CNV frequency among different CHD subtypes in the cohort were calculated and compared. The results of CMA analysis of the two cohorts were synthesised and compared with those of the normal control group. Finally, a new candidate for CHD was selected. Finally, an online analysis tool was used. Gene expression database and pathway analysis were used to identify new candidate genes for CHD. Results: 514 cases of CHD were included in this study. Of these, 422 were from BCH cohorts and 92 from SCMC queues. For BCH queues, The overall molecular diagnostic rate of CHD by clinical CMA was 12.8-18.48.The molecular diagnostic rate of CHD was 14.11-20.56 excluding 12 cases with chromosomal aneuploidy, and the diagnostic rate of CHD was 4.32-9.26 in all CHD subtypes. The frequency of pathogenetic CNV in the conical trunk malformation and cardiac septal defect was the highest. The molecular diagnostic rate of the cohort was slightly lower than 8.7-12. In addition, In this study, four chromosomal regions with recurrence times 鈮，

本文編號(hào)：1675597