一特殊角膜營養(yǎng)不良家系及一原發(fā)性開角型青光眼家系的遺傳學(xué)研究
發(fā)布時(shí)間：2018-11-10 21:30
【摘要】：目的觀察1特殊角膜營養(yǎng)不良(corneal dystrophy,CD)家系及1原發(fā)性開角型青光眼(primary open-angle glaucoma,POAG)家系的臨床表現(xiàn),同時(shí)分析和探討兩家系的基因突變。方法1.收集1 CD家系3代共13人。通過眼科常規(guī)檢查觀察其臨床表現(xiàn)并繪制家系遺傳圖譜及分析臨床表現(xiàn)特點(diǎn)。在經(jīng)患者及家系正常成員的知情同意后,對(duì)CD家系6個(gè)DNA樣本(其中4個(gè)為患者樣本),采用全基因組平均間隔約10c M左右(Marshfield遺傳圖譜)總共366個(gè)STR位點(diǎn)進(jìn)行連鎖分析及單倍型推斷,確定該家系可能致病基因所在的區(qū)域。再選擇家系2名CD患者進(jìn)行全基因外顯子測序后,獲得定位于連鎖分析確定的區(qū)域內(nèi)的候選基因。將候選基因在家系及200正常人群中進(jìn)行Sanger測序驗(yàn)證。ANNOVAR及GERP軟件被用來預(yù)測分析蛋白功能。2.收集1 POAG家系5代共23人。通過眼科常規(guī)檢查觀察其臨床表現(xiàn)并繪制家系遺傳圖譜及分析臨床表現(xiàn)特點(diǎn)。在經(jīng)患者及家系正常成員的知情同意后,選擇POAG家系的2名患者和1名正常成員的外周血DNA進(jìn)行全基因組外顯子測序。分析測序數(shù)據(jù)及篩選變異后獲得候選變異。選擇位于POAG常見致病基因連鎖定位區(qū)域內(nèi)的候選變異,通過對(duì)家系其他成員及200正常人群進(jìn)行基因靶向Sanger測序驗(yàn)證。并采用SIFT與Poly Phen軟件預(yù)測蛋白功能。結(jié)果1.在CD家系中共6名成員被診斷為CD,平均發(fā)病年齡為16.5歲。該家系的臨床特征為:開始以角膜緣內(nèi)皮及后彈力層處出現(xiàn)點(diǎn)狀、散在分布的乳白色混濁,并沿著角膜緣發(fā)展至貫通的同時(shí),向角膜內(nèi)皮中央發(fā)展�；鞚釣殡p側(cè)、對(duì)稱性表現(xiàn)。約40歲左右出現(xiàn)深基質(zhì)層混濁,并隨著年齡的增長,混濁發(fā)展至前基質(zhì)層。我們首次報(bào)道了以角膜緣內(nèi)皮層及后彈力層開始病變,并逐漸向角膜中央及淺基質(zhì)層發(fā)展為特征的CD。在6名患者的基因序列檢測中均發(fā)現(xiàn)KIAA1522基因雜合子突變(c.1331GA)。而家系非患者及200名對(duì)照者中均未發(fā)現(xiàn)該基因突變。ANNOVAR及GERP軟件均提示該突變影響蛋白功能。2.在POAG家系中,6名有嚴(yán)重臨床表現(xiàn)和高眼壓病史的成員被診斷為POAG,平均發(fā)病年齡為26.3歲,其中67%的病人需要濾過性手術(shù)治療。其他成員無臨床表現(xiàn)。在6名患者和3名無癥狀成員中,通過基因測序發(fā)現(xiàn)了myocilin蛋白基因(MYOC)中的3號(hào)外顯子的c.C1456T突變。另外,在6名患者和3名無癥狀成員中,我們也發(fā)現(xiàn)了一從未報(bào)道過的新的基因突變:β-1,4-半乳糖基轉(zhuǎn)移酶3基因(B4GALT3)中的c.G322A突變。在對(duì)照組及其他家庭成員中未發(fā)現(xiàn)這兩種突變。SIFT,Poly Phen功能預(yù)測軟件均提示兩突變影響蛋白功能。結(jié)論1.我們首次報(bào)道了以角膜緣內(nèi)皮層及后彈力層開始病變,并逐漸向角膜中央及淺基質(zhì)層發(fā)展為特征的CD。KIAA1522基因(c.1331GA)雜合子突變可能與該家系CD的發(fā)生及該病特殊的病變特征相關(guān)。2.MYOC基因的c.1456CT(p.L486F)突變和B4GALT3基因的c.322GA(p.V108I)突變可能與該P(yáng)OAG家系的發(fā)病相關(guān)。
[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.
【學(xué)位授予單位】：安徽醫(yī)科大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2016
【分類號(hào)】：R772.2;R775
本文編號(hào)：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.
【學(xué)位授予單位】：安徽醫(yī)科大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2016
【分類號(hào)】：R772.2;R775
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