臺灣博碩士論文加值系統

心臟是胚胎發育過程中最早發育的器官之一，一般認為先天性心臟病的發生與中樞神經系統之心臟神經脊 (cardiac neural crest) 發育得正常與否有密切的關係。小兒先天性心臟病 (congenital heart diseases) 每一年在台灣所造成的死亡率為兒童癌症的兩倍。探討造成先天性心臟病的致病基因與心臟發育的相關性在優生學和治療策略的發展中，將可提供早期預防、診斷或開發藥物的有效途徑。先天性心臟病發生的時間與胚胎發育早期、心臟形成有密切的關係。雖然藥物和母體環境皆可能造成先天性心臟病的發生，但家族中若有罹患先天性心臟病，在發生的比例較一般無先天性心臟並家族要高，顯示先天性心臟病與個體基因型 (genotypes) 有關。本研究利用來自高雄榮總小兒科共 239 個小兒先天性心臟病家庭總計 245 位病人。其中有 83 位病人被診斷是心室中膈缺損，為 34.7% ，所佔的比例最高。進行病人與親代血液採樣及基因組DNA之萃取，進一步利用位於第 22 對染色體十個涵蓋DiGeorge syndrome 之微衛星型 (microstallite) 遺傳標記，進行台灣小兒先天性心臟病與心臟發育相關基因型的探討。進一步進行染色體細部基因定位 (fine mapping) 與局部候選基因分析方式 (local candidate analysis) ，希望藉此研究將導致台灣小兒先天性心臟病的基因定位並縮小範圍到第 22 對染色體內較小之區域。目前結果顯示有二十五個心室中膈缺損的家族顯示在 22q11 的位置上有 loss of heterozygosity 的現象，分別在 D22S264, D22S303, D22S420, D22S427, D22S941, D22S944, D22S1638, D22S1648 發生。將進一步研究該區域與心臟發育相關的候選基因 TBX1, DGCR6, UFD1L 之基因是否有所突變。另外，利用單股結構多樣性的實驗 (single strand conformation polymorphisms) 分析其他三個位於不同染色體上但與心臟發育相關的基因 CSX, JAG1, TBX5 是否有所突變而造成心室中膈缺損。目前研究結果顯示TBX5的第四個、第五個、第九個及第十個的表現子，在245位病人與其親代間並沒有發現突變，所以我們即證明這四個表現子對於這245位小兒先天性心臟病病人的心臟發育無直接關係。但我們仍會繼續研究這三個基因其他表現子的序列是否在這些病人中是否有發生突變的情形。

Objective. Congenital heart disease (CHDs) in Taiwan cause twice as many children die each year comparing with childhood cancers. Prevalent CHDs are ventricular septal defects (VSDs) which accounted for ~40% Taiwanese population averagely. Studies on heart development-related genes on the human genome will provide valuable information for early diagnosis/prevention in eugenics and the development of therapeutic strategies.
Methods. A total of 239 CHD families from Kaohsiung Veteran General Hospital, including 713 individuals with 245 affected, participated in this study. Among these CHDs families, 83 were diagnosed as VSDs, accounted for 34.7% of all CHDs. We initiated using a semi-quantitative fluorescent PCR method applying ten highly polymorphic markers that located within 22q11, genotyping analysis for deletion or loss of heterozygosity. In those cases that are identified as chromosome 22q11-independent VSDs, cardiac development-related candidate genes TBX5, CSX and JAG1 analyses were performed by Single-Strand Conformation Polymorphisms (SSCPs) and Temporal Temperature Gradient Gel Electrophoresis (TTGE) analyses to identify whether any genomic mutation/deletion exists.
Results. So far, there are twenty-five VSD affected individuals have been identified as loss of heterozygosity (LOH) at loci D22S264, D22S303, D22S420, D22S427,D22S941, D22S944, D22S1638 and D22S1648. Candidate gene approaches will therefore be carried out within chromosome 22q11 subregion in these individuals.
Conclusions. The frequency of CHD necessitating intervention in patients referred for cardiovascular evaluation after diagnosis of a chromosome 22q11 deletion. Routine screening for CHDs, including VSD and other imaging studied to identify the any microdeletion(s) or LOH.

中文摘要......................................................Ⅰ
英文摘要......................................................Ⅲ
英文縮寫表....................................................Ⅳ
前言...........................................................1
材料與方法....................................................24
結果..........................................................31
討論..........................................................49
結論..........................................................55
參考文獻......................................................56

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