青少年开角型青光眼遗传家系MYOC基因突变的鉴定研究

发布时间：2018-04-01 03:29

本文选题：青光眼　切入点：开角型　出处：《中国人民解放军医学院》2014年博士论文

【摘要】：目的： 1.建立青少年开角型青光眼（Juvenile-Onset open-angle glaucoma,JOAG）遗传家系临床信息和数字档案资料及DNA标本库。 2.分析青少年开角型青光眼遗传家系临床表型和遗传学特征。 3.鉴定引起青少年开角型青光眼遗传家系致病基因突变位点。方法： 1.收集一个4代共14人的青少年开角型青光眼遗传家系成员病史及临床资料，签署临床调查及科研知情同意书，填写JOAG调查表，绘制家系图谱。抽取家系内12个成员外周血8～10ml并进行了系统的眼科检查，包括矫正视力，眼压测量，角膜中央区厚度测量，房角检查，裂隙灯检查，眼底检查、视野检查和视盘及黄斑OCT检查。 2.应用聚合酶链反应（polymeasreehaineraction, PCR)和Sanger测序法，对该家系12名成员4个候选基因（CYP1B1, MYOC, OPTN和WDR36基因）所有外显子区域进行致病基因突变位点筛查，结果与NCBI的dbSNP数据库、千人数据库、ensemble数据库等进行比对，并且将该突变位点在家系内无亲缘关系的正常人和100个家系之外的对照组中进行验证。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GD-JY001遗传家系临床资料和数字档案数据库及DNA标本库。 2.该家系4代共14名成员,现存活患者12名。5个患者发病年龄平均14.4岁（11-16岁），平均眼压44.2mmHg（28-60mmHg），视神经出现不对称性改变和视野损害。患者的房角检查显示小梁网有更加多的色素沉积和虹膜根粗大形成房角粘连。家族中大多患者带有一个较薄的中心角膜厚度（central corneathickness，CCT），患者的平均CCT为488.625mm（529-446mm）。 3. GD-JY001家系所有患者在MYOC基因第3号外显子编码区发现杂合错义突变c.733T＞G，p.Cys245Gly，该突变与GD-JY001家系表型共分离。在NCBI的dbSNP数据库、千人数据库、ensemble数据库以及www.myocilin.com网站中均未发现有该位点改变的报道，并且在100个无亲缘关系的正常人中不存在该突变位点。测序结果在CYP1B1, OPTN和WDR36基因的外显子区未发现致病突变。结论： 1.青少年开角型青光眼GD-JY001家系遗传方式符合常染色体显性遗传特征。患者临床表型为发病时间早，眼压升高显著，眼底视神经损害呈不对称性改变，视野损害与眼底改变一致。所有患者有异常的房角改变包括房角色素增加和虹膜根异常增粗并与房角形成粘连。 2.杂合错义突变c.733T＞G，p.Cys245Gly是青少年开角型青光眼GD-JY001家系致病基因突变，其候选基因CYP1B1, OPTN和WDR36与该家系的致病基因无相关性。
[Abstract]:Objective:. 1. To establish the pedigrees clinical information, digital file data and DNA specimen library of juvenile open angle glaucoma (juvenile open angle glaucoma). 2. To analyze the clinical phenotype and genetic characteristics of pedigrees with juvenile open angle glaucoma. 3.Identification of mutation locus of pathogenic gene in pedigrees of pedigrees causing juvenile open-angle glaucoma. Methods:. 1. To collect the medical history and clinical data of a family of 14 adolescents with open angle glaucoma for 4 generations, sign the consent form of clinical investigation and scientific research, and fill out the JOAG questionnaire. Drawing pedigree map. Peripheral blood 8~10ml was extracted from 12 members of the family and systematic ophthalmic examination was carried out, including corrected visual acuity, intraocular pressure measurement, corneal central area thickness measurement, atrial angle examination, slit lamp examination, fundus examination. Visual field examination and OCT examination of optic disc and macula. 2. Polymerase chain reaction (PCR) and Sanger sequencing were used to screen all exons of CYP1B1, MYOC, OPTN and WDR36 gene. The results were compared with the dbSNP database of NCBI. The database of thousands of people was compared, and the mutation site was verified in the normal persons who were not related to each other in the family and the control group outside 100 families. Results:. 1. The GD-JY001 genealogy clinical data database, digital file database and DNA specimen database were established, which were scientific, standardized and in accordance with the principles of genetic resources collection and research. 2. The family consists of 14 members for four generations, There were 12 surviving patients. The mean age of onset of 5 patients was 14.4 years old and 11-16 years old, the mean IOP was 44.2 mm HgGG 28-60 mm HgG, the optic nerve was asymmetrical and the visual field was damaged. The atrial angle examination of the patients showed more pigmentation of trabecular meshwork and thicker iris root. Most patients in the family had a thin central corneal thickness (CCT) with an average CCT of 488.625mm (529-446mm). 3. The heterozygous missense mutation c.733T was found in all patients of GD-JY001 pedigree in the coding region of exon 3 of MYOC gene. The mutation was coisolated from the phenotype of GD-JY001 pedigree, and was isolated from the dbSNP database of NCBI. No change of the locus was reported in the thousands of human databases and www.myocilin.com sites, and the mutation was not found in 100 unrelated normal individuals. No mutations were found in the exon region of CYP1B1, OPTN and WDR36 genes. Conclusion:. 1. The genetic pattern of GD-JY001 pedigrees in juvenile open-angle glaucoma was autosomal dominant. The clinical phenotypes were early onset, significant elevation of intraocular pressure and asymmetric changes of optic nerve damage in the fundus. All patients had abnormal atrial angle changes including increased atrial role factor and abnormal thickening of iris root and formed adhesion with the angle of atrium. 2. The heterozygous missense mutation c.733T > GPp.Cys245Gly is a mutation of GD-JY001 gene in the pedigrees of juvenile open-angle glaucoma. The candidate genes CYP1B1, OPTN and WDR36 are not correlated with the pathogenic genes of this family.
【学位授予单位】：中国人民解放军医学院
【学位级别】：博士
【学位授予年份】：2014
【分类号】：R775

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