Reis-Bucklers角膜营养不良遗传家系TGFBI基因突变研究

发布时间：2018-03-13 17:41

本文选题：颗粒状角膜营养不良　切入点：遗传　出处：《中国人民解放军医学院》2014年硕士论文　论文类型：学位论文

【摘要】：目的： 1.采集Reis-Bucklers角膜营养不良（Reis-bucklers Corneal Dystrophy，RBCD）遗传家系临床信息，建立电子档案资料及DNA标本库。 2.分析Reis-Bucklers角膜营养不良遗传家系的临床表型和遗传学特征 3.对Reis-Bucklers角膜营养不良候选基因TGFBI的所有外显子进行基因测序，确定致病基因突变位点。方法： 1.收集一个3代共18名成员的Reis-Bucklers角膜营养不良家系，并命名为HN-SY-001家系，其中9人被确诊为RBCD。采集该病史资料，填写RBCD调查表、签署参与研究相关知情同意书，绘制家系图谱。对家系内所有成员进行系统的眼部病史和系统疾病回顾，进行眼部专科检查和必要的系统检查。眼部检查包括视力、矫正视力、眼压、角膜、隙灯检查、角膜照相以及泪液分泌检查和角膜敏感性检查。 2.抽取患者及家系内正常成员外周血8-10ml，提取DNA。应用聚合酶链反应（polymeasreehaineraction，PCR）和Sanger直接测序方法，对该家系全部现存活成员的TGFBI基因的所有16个外显子区域进行致病基因突变位点的筛查。发现可疑阳性位点后，，与千人数据库和ensemble数据库等进行比对，并且将该位点在100个家系之外的无亲缘关系的正常人中进行进一步的验证。结果： 1.建立了科学、规范，符合遗传资源采集研究原则要求的Reis-Bucklers角膜营养不良遗传家系临床资料和数字档案数据库及DNA标本库。 2. HN-SY-001家系患者9例诊断为Reis-Bucklers角膜营养不良，发病年龄1~10（3.67±0.90）岁，首发症状包括畏光流泪，眼睛疼痛。视力下降年龄6~13（8.00±0.78）岁。个体患者主要表现为角膜上皮地图样缺损、浅基质层孤岛样变性混浊、角膜表面铁锈样色素沉积和深部角膜基质层水肿。所有患者角膜敏感度都有减弱甚至消失。 3. Reis-Bucklers角膜营养不良家系患者在TGFBI基因第4外显子编码区发现杂合错义突变c.418GT（p. R124L），该突变在家系内表型共分离，并在100个无亲缘关系的正常人中不存在该突变位点。结论： 1.该家系由TGFBI基因c.418GT突变引起的RBCD患者存在显著表型异质性。且和以往报道的不同，患者的水肿等病变累及全基质层。 2. Reis-Bucklers角膜营养不良家系符合常染色体显性遗传方式，TGFBI基因是该家系的致病基因，杂合错义突变c.418GT（p. R124L）是该家系的致病基因突变。
[Abstract]:Objective:. 1. To collect the clinical information of Corneal dystrophy of Reis-Bucklers Corneal dystrophytes, and to establish the electronic archives and DNA specimen library. 2. To analyze the clinical phenotypic and genetic characteristics of Reis-Bucklers corneal dystrophy. 3. All exons of Reis-Bucklers corneal dystrophy candidate gene TGFBI were sequenced to identify the mutation site of the pathogenic gene. Methods:. 1. A family of 18 members with 3 generations of Reis-Bucklers corneal dystrophy was collected and named as HN-SY-001 family. Among them, 9 of them were diagnosed as RBCD.The medical history was collected, the RBCD questionnaire was filled out, and the consent to participate in the study was signed. Family atlas. Systematic eye history and disease review of all members of the family, eye specialist examinations and necessary systematic examinations. Eye examinations include visual acuity, corrected visual acuity, intraocular pressure, cornea, lacunae, etc. Corneal photography, tear secretion and corneal sensitivity. 2. The peripheral blood of patients and normal family members were extracted from 8-10ml, and DNAs were extracted. Polymerase chain reaction (PCR) and Sanger sequencing were used. All the 16 exons of the TGFBI gene of all the surviving members of the family were screened for mutation sites of the pathogenic gene. After the suspicious positive loci were found, they were compared with the database of thousands of people and the database of ensemble, etc. The locus was further validated in unrelated normal individuals from 100 families. Results:. 1. To establish a scientific, standardized and standard Reis-Bucklers corneal dystrophy genetic pedigree clinical data base, digital archival database and DNA specimen database according to the principle of genetic resource collection and research. 2. Reis-Bucklers corneal dystrophy was diagnosed in 9 patients with HN-SY-001 pedigree. The onset age was 1.103.67 卤0.90 years old. The initial symptoms included photophobia, eye pain, visual acuity loss (6138.00 卤0.78) years old. The main symptoms of the patients were corneal epithelial-like defects. The corneal sensitivity was decreased or even disappeared in all the patients with insular degeneration and opacity of shallow stroma, rust-like pigment deposition on the corneal surface and edema in the deep corneal stroma. 3. Heterozygous mutation c. 418 GTN pR124L was found in the coding region of exon 4 of TGFBI gene in patients with Reis-Bucklers corneal dystrophy. Conclusion:. 1. The phenotypic heterogeneity of RBCD patients caused by the mutation of TGFBI gene c.418GT was significant, and different from the previous reports, the edema of the patients involved the whole stromal layer. 2. The Reis-Bucklers corneal dystrophy family is in accordance with the autosomal dominant genetic pattern. The heterozygous mutation c.418GTpR124L is the pathogenic gene mutation of this family.
【学位授予单位】：中国人民解放军医学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R772.2

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