一特殊角膜营养不良家系及一原发性开角型青光眼家系的遗传学研究
发布时间：2018-11-10 21:30
【摘要】：目的观察1特殊角膜营养不良(corneal dystrophy,CD)家系及1原发性开角型青光眼(primary open-angle glaucoma,POAG)家系的临床表现,同时分析和探讨两家系的基因突变。方法1.收集1 CD家系3代共13人。通过眼科常规检查观察其临床表现并绘制家系遗传图谱及分析临床表现特点。在经患者及家系正常成员的知情同意后,对CD家系6个DNA样本(其中4个为患者样本),采用全基因组平均间隔约10c M左右(Marshfield遗传图谱)总共366个STR位点进行连锁分析及单倍型推断,确定该家系可能致病基因所在的区域。再选择家系2名CD患者进行全基因外显子测序后,获得定位于连锁分析确定的区域内的候选基因。将候选基因在家系及200正常人群中进行Sanger测序验证。ANNOVAR及GERP软件被用来预测分析蛋白功能。2.收集1 POAG家系5代共23人。通过眼科常规检查观察其临床表现并绘制家系遗传图谱及分析临床表现特点。在经患者及家系正常成员的知情同意后,选择POAG家系的2名患者和1名正常成员的外周血DNA进行全基因组外显子测序。分析测序数据及筛选变异后获得候选变异。选择位于POAG常见致病基因连锁定位区域内的候选变异,通过对家系其他成员及200正常人群进行基因靶向Sanger测序验证。并采用SIFT与Poly Phen软件预测蛋白功能。结果1.在CD家系中共6名成员被诊断为CD,平均发病年龄为16.5岁。该家系的临床特征为:开始以角膜缘内皮及后弹力层处出现点状、散在分布的乳白色混浊,并沿着角膜缘发展至贯通的同时,向角膜内皮中央发展。混浊为双侧、对称性表现。约40岁左右出现深基质层混浊,并随着年龄的增长,混浊发展至前基质层。我们首次报道了以角膜缘内皮层及后弹力层开始病变,并逐渐向角膜中央及浅基质层发展为特征的CD。在6名患者的基因序列检测中均发现KIAA1522基因杂合子突变(c.1331GA)。而家系非患者及200名对照者中均未发现该基因突变。ANNOVAR及GERP软件均提示该突变影响蛋白功能。2.在POAG家系中,6名有严重临床表现和高眼压病史的成员被诊断为POAG,平均发病年龄为26.3岁,其中67%的病人需要滤过性手术治疗。其他成员无临床表现。在6名患者和3名无症状成员中,通过基因测序发现了myocilin蛋白基因(MYOC)中的3号外显子的c.C1456T突变。另外,在6名患者和3名无症状成员中,我们也发现了一从未报道过的新的基因突变:β-1,4-半乳糖基转移酶3基因(B4GALT3)中的c.G322A突变。在对照组及其他家庭成员中未发现这两种突变。SIFT,Poly Phen功能预测软件均提示两突变影响蛋白功能。结论1.我们首次报道了以角膜缘内皮层及后弹力层开始病变,并逐渐向角膜中央及浅基质层发展为特征的CD。KIAA1522基因(c.1331GA)杂合子突变可能与该家系CD的发生及该病特殊的病变特征相关。2.MYOC基因的c.1456CT(p.L486F)突变和B4GALT3基因的c.322GA(p.V108I)突变可能与该POAG家系的发病相关。
[Abstract]:Objective to observe the clinical manifestations of special corneal dystrophy (corneal dystrophy,CD) and primary open-angle glaucoma (primary open-angle glaucoma,POAG) pedigree. Method 1. A total of 13 CD families were collected for 3 generations. The clinical manifestation was observed by routine ophthalmology examination, the genetic map of family was drawn and the clinical manifestation was analyzed. With the informed consent of the patient and normal members of the family, six DNA samples (four of which were patient samples) from the CD family, A total of 366 STR loci with an average genome interval of about 10 cm (Marshfield genetic map) were used for linkage analysis and haplotype inference to determine the region of the family where the possible pathogenic gene was located. The candidate genes located in the region identified by linkage analysis were obtained after two CD patients were sequenced. The candidate genes were sequenced by Sanger sequencing in home line and 200 normal population. ANNOVAR and GERP software were used to predict the function of protein. 2. 2. A total of 23 individuals were collected from 5 generations of 1 POAG family. The clinical manifestation was observed by routine ophthalmology examination, the genetic map of family was drawn and the clinical manifestation was analyzed. With the informed consent of patients and normal members, the peripheral blood DNA of two patients and one normal member of POAG family were selected to sequence the whole genome exon. The candidate mutation was obtained by analyzing the sequencing data and screening the mutation. The candidate mutations located in the linkage region of common pathogenic genes of POAG were selected and confirmed by gene targeting Sanger sequencing of other family members and 200 normal population. SIFT and Poly Phen software were used to predict the function of protein. Result 1. The average age of onset of CD, in 6 members of the CD family was 16. 5 years. The clinical features of the pedigree were as follows: the corneal limbal endothelium and posterior elastic layer appeared punctate, scattered in the distribution of opacity, and developed to the center of corneal endothelium along the limbus cornea. Opacity was bilateral and symmetrical. About 40 years old, deep matrix opacification appeared, and with the age increasing, the turbidity developed to the anterior matrix layer. We report for the first time that CD., characterized by the onset of corneal limbal endothelial layer and posterior elastic layer, and gradually developed towards the central and shallow stromal layer of cornea, Heterozygote mutations (c.1331GA) of KIAA1522 gene were found in 6 patients. The gene mutation was not found in non-family patients and 200 controls. Both ANNOVAR and GERP software indicated that the mutation affected protein function. 2. In the POAG pedigree, six members with severe clinical symptoms and a history of intraocular hypertension were diagnosed with POAG, at an average age of 26.3 years, 67% of whom required filtering surgery. Other members had no clinical manifestations. In 6 patients and 3 asymptomatic members, the c.C1456T mutation in exon 3 of myocilin gene (MYOC) was identified by gene sequencing. In addition, we also found a new gene mutation in 6 patients and 3 asymptomatic members: the c.G322A mutation in the 尾-1 4-galactosyltransferase 3 (B4GALT3) gene. These two mutations were not found in the control group and other family members. The SIFT,Poly Phen function prediction software showed that the two mutations affected the protein function. Conclusion 1. We reported for the first time that the lesions began in the limbal endothelium and the posterior elastic layer. The heterozygote mutation of CD.KIAA1522 gene (c.1331GA), which gradually develops to the central and superficial stroma of cornea, may be related to the occurrence of CD and the special pathological features of the disease in this family. C.1456CT (p.L486F) of 2.MYOC gene ) mutation and c.322GA (p.V108I) mutation of B4GALT3 gene may be associated with the pathogenesis of the POAG pedigree.
【学位授予单位】：安徽医科大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R772.2;R775
本文编号：2323656
[Abstract]:Objective to observe the clinical manifestations of special corneal dystrophy (corneal dystrophy,CD) and primary open-angle glaucoma (primary open-angle glaucoma,POAG) pedigree. Method 1. A total of 13 CD families were collected for 3 generations. The clinical manifestation was observed by routine ophthalmology examination, the genetic map of family was drawn and the clinical manifestation was analyzed. With the informed consent of the patient and normal members of the family, six DNA samples (four of which were patient samples) from the CD family, A total of 366 STR loci with an average genome interval of about 10 cm (Marshfield genetic map) were used for linkage analysis and haplotype inference to determine the region of the family where the possible pathogenic gene was located. The candidate genes located in the region identified by linkage analysis were obtained after two CD patients were sequenced. The candidate genes were sequenced by Sanger sequencing in home line and 200 normal population. ANNOVAR and GERP software were used to predict the function of protein. 2. 2. A total of 23 individuals were collected from 5 generations of 1 POAG family. The clinical manifestation was observed by routine ophthalmology examination, the genetic map of family was drawn and the clinical manifestation was analyzed. With the informed consent of patients and normal members, the peripheral blood DNA of two patients and one normal member of POAG family were selected to sequence the whole genome exon. The candidate mutation was obtained by analyzing the sequencing data and screening the mutation. The candidate mutations located in the linkage region of common pathogenic genes of POAG were selected and confirmed by gene targeting Sanger sequencing of other family members and 200 normal population. SIFT and Poly Phen software were used to predict the function of protein. Result 1. The average age of onset of CD, in 6 members of the CD family was 16. 5 years. The clinical features of the pedigree were as follows: the corneal limbal endothelium and posterior elastic layer appeared punctate, scattered in the distribution of opacity, and developed to the center of corneal endothelium along the limbus cornea. Opacity was bilateral and symmetrical. About 40 years old, deep matrix opacification appeared, and with the age increasing, the turbidity developed to the anterior matrix layer. We report for the first time that CD., characterized by the onset of corneal limbal endothelial layer and posterior elastic layer, and gradually developed towards the central and shallow stromal layer of cornea, Heterozygote mutations (c.1331GA) of KIAA1522 gene were found in 6 patients. The gene mutation was not found in non-family patients and 200 controls. Both ANNOVAR and GERP software indicated that the mutation affected protein function. 2. In the POAG pedigree, six members with severe clinical symptoms and a history of intraocular hypertension were diagnosed with POAG, at an average age of 26.3 years, 67% of whom required filtering surgery. Other members had no clinical manifestations. In 6 patients and 3 asymptomatic members, the c.C1456T mutation in exon 3 of myocilin gene (MYOC) was identified by gene sequencing. In addition, we also found a new gene mutation in 6 patients and 3 asymptomatic members: the c.G322A mutation in the 尾-1 4-galactosyltransferase 3 (B4GALT3) gene. These two mutations were not found in the control group and other family members. The SIFT,Poly Phen function prediction software showed that the two mutations affected the protein function. Conclusion 1. We reported for the first time that the lesions began in the limbal endothelium and the posterior elastic layer. The heterozygote mutation of CD.KIAA1522 gene (c.1331GA), which gradually develops to the central and superficial stroma of cornea, may be related to the occurrence of CD and the special pathological features of the disease in this family. C.1456CT (p.L486F) of 2.MYOC gene ) mutation and c.322GA (p.V108I) mutation of B4GALT3 gene may be associated with the pathogenesis of the POAG pedigree.
【学位授予单位】：安徽医科大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R772.2;R775
【参考文献】
相关期刊论文 前8条
1 程玄;秦丽敏;刘铁城;刘伟;陈丽丽;张译心;;一个Reis-Bucklers角膜营养不良遗传家系的TGFBI基因突变研究[J];解放军医学杂志;2014年06期
2 刘玉莲;李璇;卢建民;邹鹏飞;;斑状角膜营养不良的临床特征及组织病理学改变[J];眼科新进展;2013年11期
3 Su-Juan Zhao;Ya-Nan Zhu;Xing-Chao Shentu;Qi Miao;;Chinese family with atypical granular corneal dystrophy type I caused by the typical R555W mutation in TGFBI[J];International Journal of Ophthalmology(English Edition);2013年04期
4 王娟娟;廖荣丰;;Avellino型角膜营养不良一家系基因研究[J];安徽医科大学学报;2013年06期
5 杨晓波;陈丽娟;;上皮基底膜角膜营养不良家系的TGFBI基因突变1例研究[J];国际眼科杂志;2012年10期
6 ;Arg124Cys mutation of the TGFBI gene in a Chinese pedigree of Reis-Bücklers corneal dystrophy[J];International Journal of Ophthalmology(English Edition);2011年03期
7 霍亚楠;姚玉峰;;胶滴状角膜营养不良的基因诊断及治疗新进展[J];浙江大学学报(医学版);2006年02期
8 葛坚,卓业鸿,郭彦,明文玉,银巍;Gene mutation in patients with primary open angle glaucoma in a pedigree in China[J];Chinese Medical Journal;2000年03期
相关博士学位论文 前1条
1 解智慧;一种新的介导细胞间粘附的肌动蛋白相关蛋白KIAA1522在食管癌中作用及其机制的研究[D];北京协和医学院;2014年
，本文编号：2323656
本文链接：https://www.wllwen.com/yixuelunwen/wuguanyixuelunwen/2323656.html
最近更新
教材专著
