目的:分析不同浓度重组myocilin蛋白对体外培养的原发性开角型青光眼(primary open angle glaucoma,POAG)患者小梁网细胞纤维连接蛋白(fibronectin,FN)表达及对黏附功能的影响。方法:体外培养POAG患者小梁网细胞,分别加入含重组myocili...目的:分析不同浓度重组myocilin蛋白对体外培养的原发性开角型青光眼(primary open angle glaucoma,POAG)患者小梁网细胞纤维连接蛋白(fibronectin,FN)表达及对黏附功能的影响。方法:体外培养POAG患者小梁网细胞,分别加入含重组myocilin蛋白终浓度为0(对照组)、0.5、1、1.5、2、2.5μg/mL的无血清培养基,运用Western blot、ELISA法分别检测小梁网细胞内和细胞培养液中FN的表达水平;并运用CCK-8法检测myocilin蛋白对POAG小梁网细胞黏附的影响。结果:Western blot检测结果:FN/β-actin比值分别为34.8±0.6、33.4±1.0、28.9±0.8、21.6±0.9、15.9±1.1、11.9±0.8;0.5μg/mL组与对照组相比,差异无统计学意义(P=0.092);1.0、1.5、2、2.5μg/mL组与对照组比较,差异有统计学意义(F=346.131,P<0.05)。ELISA法检测各组细胞培养液中FN浓度为:0.4654±0.0039、0.4596±0.0032、0.4216±0.0037、0.4214±0.0039、0.4043±0.0039、0.3806±0.0071μg/mL;0.5μg/mL组与对照组相比,差异无统计学意义(P=0.120);1.0、1.5、2、2.5μg/mL组与对照组比较,差异有统计学意义(F=176.054,P<0.05)。运用CCK-8法检测各组细胞吸光度均值分别为1.98140.1624、1.8848±0.0267、1.4895±0.0916、1.4120±0.1087、1.3392±0.1391、1.0310±0.0639;0.5μg/mL组与对照组相比,差异无统计学意义(P=0.300);1、1.5、2、2.5μg/mL组与对照组相比,差异有统计学意义(F=177.818,P<0.05)。结论:Myocilin蛋白能抑制POAG小梁网细胞FN的表达,并呈现一定的剂量依赖性;myocilin蛋白可降低POAG小梁网细胞的黏附能力。展开更多
AIM: To describe the anticipation and anti-glaucoma drugs response of a Chinese family with juvenile-onset open angle glaucoma(JOAG) caused by the Pro370Leu myocilin(MYOC) mutation. ·METHODS: Fifteen members of a...AIM: To describe the anticipation and anti-glaucoma drugs response of a Chinese family with juvenile-onset open angle glaucoma(JOAG) caused by the Pro370Leu myocilin(MYOC) mutation. ·METHODS: Fifteen members of a three-generation Chinese family with JOAG were recruited to this study. They all underwent ophthalmic common examinations. Patients suspected to have JOAG got an assessment of visual field and optical coherence tomography. Intraocular pressures(IOPs) of four patients were measured at 8,10,12,14,17 o’clock respectively after using anti-glaucoma drugs. Mutation screening of all MYOC gene coding exons of the participants was performed by using direct sequencing of PCR products. ·RESULTS: Clinical examinations and pedigree analysis revealed eight family members were suffered from JOAG. Apparent genetics anticipation phenomenon was observed in this family. Their clinical features included elevated IOP of 35-55mmHg,loss of visual field,thinning of retinal nerve fiber layer,and glaucomatous optic disc damage. Noticeably,their intraocular pressure levels could be controlled within normal range at 8 and 10 o’clock by anti-glaucoma drugs,but their IOPs would elevate 】21mmHg after 12 o’clock. Seven patients received trabeculectomy produced thin-walled,pale,and saccate filtering blebs maintaining lower intraocular pressure efficiently. Mutation screening indentified aheterozygous C→T missense mutation in the MYOC gene at position 1 109 in exon 3,corresponding to a substitution of a highly conserved proline to leucine at codon 370 in the olfactomedin domain of MYOC. ·CONCLUSION: The clinical characteristics of JOAG in this family were 1) genetics anticipation; 2) high IOP; 3) temporay response to anti-glaucoma drugs; 4) filtering surgery produced thin-walled and saccate filtering blebs,helping maintain lower IOP.展开更多
Background: Glaucoma is the leading cause of irreversible blindness incapacitating over 80 million people worldwide. Several pathogenetic mechanisms have been postulated to explain the optic nerve damage that occur in...Background: Glaucoma is the leading cause of irreversible blindness incapacitating over 80 million people worldwide. Several pathogenetic mechanisms have been postulated to explain the optic nerve damage that occur in POAG among which genetic predisposition is prominent. Gene-Linkage-based studies have identified genes associated with POAG: Myocilin, Optineurin, WDR36, Tank-Binding Kinase (TBK1) and APbb-2. Objective: To investigate the prevalence of myocilin gene mutation in adult-onset POAG patients and non-glaucoma subjects who are indigenes of Rivers State. Methodology: In this comparative cross-sectional study, 393 POAG patients attending the Glaucoma Clinic of UPTH were compared with 393 age and sex-matched phenotypically normal participants. Clinical assessment combined with findings from clinical records was used. Venous blood was obtained for genomic analyses. Extracted DNA was sequenced with specific primers for myocilin and polymerase chain reaction. Zymo-Bead Genomic DNA kit protocol was used to detect allelic differences. Results: Total of 786 participants participated in the study. The mean age was 59.8 ± 11.8 years. The prevalence of myocilin gene mutation (MYOC) in the study population was 5.3%, in the POAG group was 8.4%, and 2.3% in the non-glaucoma group. This observed difference was statistically significant (p = 0.001). Location of the mutant myocilin gene was in GLC1A 171638779, 171638703, 171638610 and 171638608. Conclusion: Mutations in myocilin gene are associated with adult-onset POAG in Rivers State. Its relevance as a biomarker for diagnosis of adult-onset POAG needs further investigations.展开更多
Background:Pathogenic mechanisms leading to open-angle glaucoma(OAG)are genetically complex.They involve neuroinflammation,elevation of intra-ocular pressures(IOP)and optic nerve hypersensitivity to cellular stresses....Background:Pathogenic mechanisms leading to open-angle glaucoma(OAG)are genetically complex.They involve neuroinflammation,elevation of intra-ocular pressures(IOP)and optic nerve hypersensitivity to cellular stresses.We mapped a locus at chromosome 20q13 that contains a modifier gene for glaucoma severity.While searching for its identity,we named this gene modifier of glaucoma 1(MOG1).The goal of this study is to characterize the mechanism by which MOG1 delays the age of onset of glaucoma when OAG is caused by mutations in the MYOC gene.We hypothesized that MOG1 mechanism may be linked to a specific endophenotype and thus dissected ocular phenotypes present in a large French-Canadian MYOC glaucoma pedigree.Methods:We studied 375 members of the CA pedigree in which autosomal dominant OAG is caused by the MYOCK423E mutation.In this family,wild-type MOG1(normal form)delays the age-at-onset(AAO)of glaucoma.Ocular records of MYOCK423E carriers were reviewed to extract the values of four quantitative traits portraying four endophenotypes:(I)age of maximal intra-ocular pressure(IOP max),(II)IOP progression,(III)rate of optic nerve degeneration and,(IV)AAO defined as the age at which IOP≥22 mmHg or age at which optic disk degeneration was first detected.Endophenotypes were tested for their heritability.A three-stage algorithm was designed to detect double mutants who carry the MYOCK423E mutation and putative MOG1 mutations.Quantitative traits values of double-mutants were then compared.Results:We found 156 individuals who were heterozygotes(HTZ)for MYOCK423E.One hundred and twenty of these were classified affected as they were OAG or had treatment for ocular hypertension(OHT)with IOP≥22 mmHg.The other 36 HTZ were asymptomatic.Only two endophenotypes,AAO and IOP max,showed significant heritability.OHT was the 1st symptom detected in 99%of the affecteds;it always preceded optic nerve damage.AAO of the affecteds ranged from 7 to 63 years old while rates of optic nerve degeneration did not significantly change between them.When comparing the AAOs of the double mutants(those who are MYOCK423E HTZ+MOG1 mutant)with the median AAOs of their respective neighbors(≤1st cousins)who are MYOCK423E HTZ and MOG1 wild-type(called single mutant as they carry a normal MOG1),we observed that the ages-at-onset of OHT in the double mutants were on average 8 years younger than the median of AAOs in their respective single mutant neighbors.Conclusions:These findings demonstrate that age-at-onset(AAO)is a reliable endophenotype to use for discovering the effect of putative MOG1 mutations in MYOCK423E carriers.They also show that the wild-type form of MOG1 delays the AAO of myocilin-induced glaucoma by about 8 years.Our study further suggests that wild-type MOG1 acts on intra-ocular pressures(IOP)by counteracting ocular hypertension(OHT)caused by mutant myocilin proteins before the beginning of optic nerve degeneration.展开更多
目的探讨一个中国人原发性开角型青光眼（primary open angle glaucoma,POAG）家系的my-ocilin基因缺陷。方法对一个5代POAG大家系进行全面的临床检查后,用聚合酶链反应扩增家系成员myocilin基因的所有外显子以及相邻部分内含子,对其产...目的探讨一个中国人原发性开角型青光眼（primary open angle glaucoma,POAG）家系的my-ocilin基因缺陷。方法对一个5代POAG大家系进行全面的临床检查后,用聚合酶链反应扩增家系成员myocilin基因的所有外显子以及相邻部分内含子,对其产物直接测序。结果家系的遗传方式符合常染色体显性遗传。确诊年龄在26～59岁之间。在所有青光眼患者,可疑者以及4例尚未出现明显青光眼体征的家系成员发现携带myocilin基因T455K突变。无该突变的家系成员中无POAG患者及可疑者。突变位于myocilin蛋白C-末端非常保守的氨基酸残基。结论T455K突变是中国人所特有的新的致病性myocilin基因突变。突变的临床表型为混合发病年龄型POAG且具有很高的外显率。这个新基因突变的发现证实中国人的致病性myocilin基因突变类型与其他种族不同。展开更多
Background： Glaucoma is a major cause of irreversible blindness worldwide. There is evidence showing that a subset of the disease is genetically determined. In this study, we screened for mutations in chromosome lq-l...Background： Glaucoma is a major cause of irreversible blindness worldwide. There is evidence showing that a subset of the disease is genetically determined. In this study, we screened for mutations in chromosome lq-linked open-angle glaucoma （GLC1A） in a Chinese family with primary open-angle glaucoma （POAG）. Methods： A total of 23 members from five generations of a family were enrolled and underwent thorough ophthalmologic examinations. In addition, 200 unrelated healthy Chinese controls were also recruited as normal control. GLC1A gene was amplified by polymerase chain reaction, and DNA sequencing was performed to screen for mutations. Results： Six members were diagnosed as POAG, with severe clinical manifestations, and history of high intraocular pressures. The mean age of disease onset was 26.3 years. However, the others were asymptomatic. In six affected and three asymptomatic members, gene sequencing revealed a mutation c.C1456T in exon 3 of myocilin gene （MYOC）. Furthermore, we also identified a novel mutation c.G322A in beta- 1,4-galactosyltransferase 3 （B4GALT3） gene in all six affected and three asymptomatic members, which was not reported previously in POAG patients. The two newly identified variants were absent in other family members as well as controls. Conclusion： The mutations c.1456C〈T （p.L486F） in MYOC and c.322G〈A （p.V1081） in B4GALT3 are likely responsible for the pathogenesis of POAG in this family.展开更多
INTRODUCTION Glaucoma is a chronic progressive neurodegenerative disease and exhibits heterogeneity, polygenic inheritance, and incomplete penetrance. Primary open-angle glaucoma （POAG） is the most common form of gl...INTRODUCTION Glaucoma is a chronic progressive neurodegenerative disease and exhibits heterogeneity, polygenic inheritance, and incomplete penetrance. Primary open-angle glaucoma （POAG） is the most common form of glaucoma. Although the underlying etiology of POAG is unknown, there is evidence that genetic mutations are closely associated with this disease, Among myocilin （MYOC） mutations, Pro370Leu （P370L） is responsible for one of the most severe glaucoma phenotypes, However, the function of MYOC protein is still not well understood.展开更多
基金Medical Scientific Research Foundation of Guangdong Province(No.A2013215)
文摘AIM: To describe the anticipation and anti-glaucoma drugs response of a Chinese family with juvenile-onset open angle glaucoma(JOAG) caused by the Pro370Leu myocilin(MYOC) mutation. ·METHODS: Fifteen members of a three-generation Chinese family with JOAG were recruited to this study. They all underwent ophthalmic common examinations. Patients suspected to have JOAG got an assessment of visual field and optical coherence tomography. Intraocular pressures(IOPs) of four patients were measured at 8,10,12,14,17 o’clock respectively after using anti-glaucoma drugs. Mutation screening of all MYOC gene coding exons of the participants was performed by using direct sequencing of PCR products. ·RESULTS: Clinical examinations and pedigree analysis revealed eight family members were suffered from JOAG. Apparent genetics anticipation phenomenon was observed in this family. Their clinical features included elevated IOP of 35-55mmHg,loss of visual field,thinning of retinal nerve fiber layer,and glaucomatous optic disc damage. Noticeably,their intraocular pressure levels could be controlled within normal range at 8 and 10 o’clock by anti-glaucoma drugs,but their IOPs would elevate 】21mmHg after 12 o’clock. Seven patients received trabeculectomy produced thin-walled,pale,and saccate filtering blebs maintaining lower intraocular pressure efficiently. Mutation screening indentified aheterozygous C→T missense mutation in the MYOC gene at position 1 109 in exon 3,corresponding to a substitution of a highly conserved proline to leucine at codon 370 in the olfactomedin domain of MYOC. ·CONCLUSION: The clinical characteristics of JOAG in this family were 1) genetics anticipation; 2) high IOP; 3) temporay response to anti-glaucoma drugs; 4) filtering surgery produced thin-walled and saccate filtering blebs,helping maintain lower IOP.
文摘Background: Glaucoma is the leading cause of irreversible blindness incapacitating over 80 million people worldwide. Several pathogenetic mechanisms have been postulated to explain the optic nerve damage that occur in POAG among which genetic predisposition is prominent. Gene-Linkage-based studies have identified genes associated with POAG: Myocilin, Optineurin, WDR36, Tank-Binding Kinase (TBK1) and APbb-2. Objective: To investigate the prevalence of myocilin gene mutation in adult-onset POAG patients and non-glaucoma subjects who are indigenes of Rivers State. Methodology: In this comparative cross-sectional study, 393 POAG patients attending the Glaucoma Clinic of UPTH were compared with 393 age and sex-matched phenotypically normal participants. Clinical assessment combined with findings from clinical records was used. Venous blood was obtained for genomic analyses. Extracted DNA was sequenced with specific primers for myocilin and polymerase chain reaction. Zymo-Bead Genomic DNA kit protocol was used to detect allelic differences. Results: Total of 786 participants participated in the study. The mean age was 59.8 ± 11.8 years. The prevalence of myocilin gene mutation (MYOC) in the study population was 5.3%, in the POAG group was 8.4%, and 2.3% in the non-glaucoma group. This observed difference was statistically significant (p = 0.001). Location of the mutant myocilin gene was in GLC1A 171638779, 171638703, 171638610 and 171638608. Conclusion: Mutations in myocilin gene are associated with adult-onset POAG in Rivers State. Its relevance as a biomarker for diagnosis of adult-onset POAG needs further investigations.
文摘Background:Pathogenic mechanisms leading to open-angle glaucoma(OAG)are genetically complex.They involve neuroinflammation,elevation of intra-ocular pressures(IOP)and optic nerve hypersensitivity to cellular stresses.We mapped a locus at chromosome 20q13 that contains a modifier gene for glaucoma severity.While searching for its identity,we named this gene modifier of glaucoma 1(MOG1).The goal of this study is to characterize the mechanism by which MOG1 delays the age of onset of glaucoma when OAG is caused by mutations in the MYOC gene.We hypothesized that MOG1 mechanism may be linked to a specific endophenotype and thus dissected ocular phenotypes present in a large French-Canadian MYOC glaucoma pedigree.Methods:We studied 375 members of the CA pedigree in which autosomal dominant OAG is caused by the MYOCK423E mutation.In this family,wild-type MOG1(normal form)delays the age-at-onset(AAO)of glaucoma.Ocular records of MYOCK423E carriers were reviewed to extract the values of four quantitative traits portraying four endophenotypes:(I)age of maximal intra-ocular pressure(IOP max),(II)IOP progression,(III)rate of optic nerve degeneration and,(IV)AAO defined as the age at which IOP≥22 mmHg or age at which optic disk degeneration was first detected.Endophenotypes were tested for their heritability.A three-stage algorithm was designed to detect double mutants who carry the MYOCK423E mutation and putative MOG1 mutations.Quantitative traits values of double-mutants were then compared.Results:We found 156 individuals who were heterozygotes(HTZ)for MYOCK423E.One hundred and twenty of these were classified affected as they were OAG or had treatment for ocular hypertension(OHT)with IOP≥22 mmHg.The other 36 HTZ were asymptomatic.Only two endophenotypes,AAO and IOP max,showed significant heritability.OHT was the 1st symptom detected in 99%of the affecteds;it always preceded optic nerve damage.AAO of the affecteds ranged from 7 to 63 years old while rates of optic nerve degeneration did not significantly change between them.When comparing the AAOs of the double mutants(those who are MYOCK423E HTZ+MOG1 mutant)with the median AAOs of their respective neighbors(≤1st cousins)who are MYOCK423E HTZ and MOG1 wild-type(called single mutant as they carry a normal MOG1),we observed that the ages-at-onset of OHT in the double mutants were on average 8 years younger than the median of AAOs in their respective single mutant neighbors.Conclusions:These findings demonstrate that age-at-onset(AAO)is a reliable endophenotype to use for discovering the effect of putative MOG1 mutations in MYOCK423E carriers.They also show that the wild-type form of MOG1 delays the AAO of myocilin-induced glaucoma by about 8 years.Our study further suggests that wild-type MOG1 acts on intra-ocular pressures(IOP)by counteracting ocular hypertension(OHT)caused by mutant myocilin proteins before the beginning of optic nerve degeneration.
文摘目的探讨一个中国人原发性开角型青光眼（primary open angle glaucoma,POAG）家系的my-ocilin基因缺陷。方法对一个5代POAG大家系进行全面的临床检查后,用聚合酶链反应扩增家系成员myocilin基因的所有外显子以及相邻部分内含子,对其产物直接测序。结果家系的遗传方式符合常染色体显性遗传。确诊年龄在26～59岁之间。在所有青光眼患者,可疑者以及4例尚未出现明显青光眼体征的家系成员发现携带myocilin基因T455K突变。无该突变的家系成员中无POAG患者及可疑者。突变位于myocilin蛋白C-末端非常保守的氨基酸残基。结论T455K突变是中国人所特有的新的致病性myocilin基因突变。突变的临床表型为混合发病年龄型POAG且具有很高的外显率。这个新基因突变的发现证实中国人的致病性myocilin基因突变类型与其他种族不同。
文摘Background： Glaucoma is a major cause of irreversible blindness worldwide. There is evidence showing that a subset of the disease is genetically determined. In this study, we screened for mutations in chromosome lq-linked open-angle glaucoma （GLC1A） in a Chinese family with primary open-angle glaucoma （POAG）. Methods： A total of 23 members from five generations of a family were enrolled and underwent thorough ophthalmologic examinations. In addition, 200 unrelated healthy Chinese controls were also recruited as normal control. GLC1A gene was amplified by polymerase chain reaction, and DNA sequencing was performed to screen for mutations. Results： Six members were diagnosed as POAG, with severe clinical manifestations, and history of high intraocular pressures. The mean age of disease onset was 26.3 years. However, the others were asymptomatic. In six affected and three asymptomatic members, gene sequencing revealed a mutation c.C1456T in exon 3 of myocilin gene （MYOC）. Furthermore, we also identified a novel mutation c.G322A in beta- 1,4-galactosyltransferase 3 （B4GALT3） gene in all six affected and three asymptomatic members, which was not reported previously in POAG patients. The two newly identified variants were absent in other family members as well as controls. Conclusion： The mutations c.1456C〈T （p.L486F） in MYOC and c.322G〈A （p.V1081） in B4GALT3 are likely responsible for the pathogenesis of POAG in this family.
基金The work was supported by a grant from Project of Science Foundation of Mudanjiang （No. G2012s0028）.
文摘INTRODUCTION Glaucoma is a chronic progressive neurodegenerative disease and exhibits heterogeneity, polygenic inheritance, and incomplete penetrance. Primary open-angle glaucoma （POAG） is the most common form of glaucoma. Although the underlying etiology of POAG is unknown, there is evidence that genetic mutations are closely associated with this disease, Among myocilin （MYOC） mutations, Pro370Leu （P370L） is responsible for one of the most severe glaucoma phenotypes, However, the function of MYOC protein is still not well understood.
