Two novel variants in CYP1B1 gene: a major contributor of autosomal recessive primary congenital glaucoma with allelic heterogeneity in Pakistani patients
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摘要
AIM: To find the CYP1 B1 mutations associated with primary congenital glaucoma(PCG) in Pakistani consanguineous pedigrees. METHODS: After getting informed consent, 11 consanguineous pedigrees belonging to different ethnic groups were enrolled. Detailed medical history was recorded and pedigrees were drawn. The standard ophthalmological examination was done to characterize the phenotype. Genomic DNA was extracted from 10 mL whole blood and coding exons and exon intron boundaries of CYP1 B1 gene were directly sequenced. Bioinformatics tools were used to model the mutant protein and predict the effect of novel variants on protein structure and function. RESULTS: Sequencing analysis revealed 5 different CYP1 B1 variants in 7 families(7/11; 64%), including two novel variants. A common mutation, p.R390 H was found in four families, whereas p.P437 L was found once in a family. Two novel variants, a homozygous non sense variant p.L13* and a compound heterozygous variant, p.P350 T along with p.V364 M were segregating with PCGin two families. All the patients had the variable onset and severity of the disease. The success rate of early clinical interventions was observed dependent on mutation types and position. Two different haplotypes were associated with frequently found mutation, p.R390 H. CONCLUSION: Identification of novel CYP1 B1 variants reassert the genetic heterogeneity of Pakistani PCG patients. The patients with missense mutations show severe phenotypic presentations and poor vision after surgical interventions as compare to patients with null variants. This may help to better understand the role of CYP1 B1 mutations in the development of PCG and its course of pathogenicity.
AIM: To find the CYP1B1 mutations associated with primary congenital glaucoma(PCG) in Pakistani consanguineous pedigrees.METHODS: After getting informed consent, 11 consanguineous pedigrees belonging to different ethnic groups were enrolled. Detailed medical history was recorded and pedigrees were drawn. The standard ophthalmological examination was done to characterize the phenotype. Genomic DNA was extracted from 10 mL whole blood and coding exons and exon intron boundaries of CYP1B1 gene were directly sequenced. Bioinformatics tools were used to model the mutant protein and predict the effect of novel variants on protein structure and function.RESULTS: Sequencing analysis revealed 5 different CYP1B1 variants in 7 families(7/11; 64%), including two novel variants. A common mutation, p.R390 H was found in four families, whereas p.P437 L was found once in a family. Two novel variants, a homozygous non sense variant p.L13* and a compound heterozygous variant, p.P350 T along with p.V364 M were segregating with PCGin two families. All the patients had the variable onset and severity of the disease. The success rate of early clinical interventions was observed dependent on mutation types and position. Two different haplotypes were associated with frequently found mutation, p.R390 H.CONCLUSION: Identification of novel CYP1B1 variants reassert the genetic heterogeneity of Pakistani PCG patients. The patients with missense mutations show severe phenotypic presentations and poor vision after surgical interventions as compare to patients with null variants. This may help to better understand the role of CYP1B1 mutations in the development of PCG and its course of pathogenicity.
AIM: To find the CYP1B1 mutations associated with primary congenital glaucoma(PCG) in Pakistani consanguineous pedigrees.METHODS: After getting informed consent, 11 consanguineous pedigrees belonging to different ethnic groups were enrolled. Detailed medical history was recorded and pedigrees were drawn. The standard ophthalmological examination was done to characterize the phenotype. Genomic DNA was extracted from 10 mL whole blood and coding exons and exon intron boundaries of CYP1B1 gene were directly sequenced. Bioinformatics tools were used to model the mutant protein and predict the effect of novel variants on protein structure and function.RESULTS: Sequencing analysis revealed 5 different CYP1B1 variants in 7 families(7/11; 64%), including two novel variants. A common mutation, p.R390 H was found in four families, whereas p.P437 L was found once in a family. Two novel variants, a homozygous non sense variant p.L13* and a compound heterozygous variant, p.P350 T along with p.V364 M were segregating with PCGin two families. All the patients had the variable onset and severity of the disease. The success rate of early clinical interventions was observed dependent on mutation types and position. Two different haplotypes were associated with frequently found mutation, p.R390 H.CONCLUSION: Identification of novel CYP1B1 variants reassert the genetic heterogeneity of Pakistani PCG patients. The patients with missense mutations show severe phenotypic presentations and poor vision after surgical interventions as compare to patients with null variants. This may help to better understand the role of CYP1B1 mutations in the development of PCG and its course of pathogenicity.
引文
1 Pascolini D,Mariotti SP.Global estimates of visual impairment:2010.Br J Ophthalmol 2012;96(5):614-618.
2 Ho CL,Walton DS.Primary congenital glaucoma:2004 update.JPediatr Ophthalmol Strabismus 2004;41(5):271-288;quiz 300-1.
3 Berraho A,Serrou A,Fritez N,El Annas A,Bencherifa F,Gaboun F,Hilal L.Genotype-phenotype correlation in Moroccan patients with primary congenital glaucoma.J Glaucoma 2015;24(4):297-305.
4 Morales-Fernandez L,Martinez-De-la-casa JM,Garcia-Bella J,Mendez C,Saenz-Frances F,Garcia M,Escribano J,Garcia-Feijoo J.Clinical variability of primary congenital glaucoma in a Spanish family with Cyp1b1 gene mutations.J Glaucoma 2015;24(8):630-634.
5 Moore DB,Tomkins O,Ben-Zion I.A review of primary congenital glaucoma in the developing world.Surv Ophthalmol2013;58(3):278-285.
6 Bashir R,Sanai M,Azeem A,Altaf I,Saleem F,Naz S.Contribution of GLC3A locus to primary congenital glaucoma in Pakistani population.Pak J Med Sci 2014;30(6):1341-1345.
7 Bejjani BA,Stockton DW,Lewis RA,Tomey KF,Dueker DK,Jabak M,Astle WF,Lupski JR.Multiple CYP1B1 mutations and incomplete penetrance in an inbred population segregating primary congenital glaucoma suggest frequent de novo events and a dominant modifier locus.Hum Mol Genet 2000;9(3):367-374.
8 Reis LM,Tyler RC,Weh E,Hendee KE,Schilter KF,Phillips JA 3rd,Sequeira S,Schinzel A,Semina EV.Whole exome sequencing identifies multiple diagnoses in congenital glaucoma with systemic anomalies.Clin Genet 2016;90(4):378-382.
9 Souma T,Tompson SW,Thomson BR,et al.Angiopoietin receptor TEKmutations underlie primary congenital glaucoma with variable expressivity.J Clin Invest 2016;126(7):2575-2587.
10 Souzeau E,Hayes M,Ruddle JB,Elder JE,Staffieri SE,Kearns LS,Mackey DA,Zhou T,Ridge B,Burdon KP,Dubowsky A,Craig JE.CYP1B1 copy number variation is not a major contributor to primary congenital glaucoma.Mol Vis 2015;21:160-164.
11 Sheikh SA,Waryah AM,Narsani AK,Shaikh H,Gilal IA,Shah K,Qasim M,Memon AI,Kewalramani P,Shaikh N.Mutational spectrum of the CYP1B1 gene in Pakistani patients with primary congenital glaucoma:novel variants and genotype-phenotype correlations.Mol Vis2014;20:991-1001.
12 Grimberg J,Nawoschik S,Belluscio L,McKee R,Turck A,Eisenberg A.A simple and efficient non-organic procedure for the isolation of genomic DNA from blood.Nucleic Acids Res 1989;17(20):8390.
13 Waryah AM,Narsani AK,Sheikh SA,Shaikh H,Shahani MY.The novel heterozygous Thr377Arg MYOC mutation causes severe Juvenile Open Angle Glaucoma in a large Pakistani family.Gene 2013;528(2):356-359.
14 Ohtake Y,Kubota R,Tanino T,Miyata H,Mashima Y.Novel compound heterozygous mutations in the cytochrome P4501B1 gene(CYP1B1)in a Japanese patient with primary congenital glaucoma.Ophthalmic Genet 2000;21(3):191-193.
15 Chen L,Huang L,Zeng A,He J.CYP1B1 gene mutations with incomplete penetrance in a Chinese pedigree with primary congenital glaucoma:a case report and review of literatures.Int J Clin Exp Med2015;8(8):14538-14541.
16 Stoilov IR,Costa VP,Vasconcellos JP,Melo MB,Betinjane AJ,Carani JC,Oltrogge EV,Sarfarazi M.Molecular genetics of primary congenital glaucoma in Brazil.Invest Ophthalmol Vis Sci 2002;43(6):1820-1827.
17 Rauf B,Irum B,Kabir F,Firasat S,Naeem MA,Khan SN,Husnain T,Riazuddin S,Akram J,Riazuddin SA.A spectrum of CYP1B1 mutations associated with primary congenital glaucoma in families of Pakistani descent.Hum Genome Var 2016;3:16021.
18 Chitsazian F,Tusi BK,Elahi E,et al.CYP1B1 mutation profile of Iranian primary congenital glaucoma patients and associated haplotypes.JMol Diagn 2007;9(3):382-393.
19 Tanwar M,Dada T,Sihota R,Das TK,Yadav U,Dada R.Mutation spectrum of CYP1B1 in North Indian congenital glaucoma patients.Mol Vis 2009;15:1200-1209.
20 Li N,Zhou Y,Du L,Wei M,Chen X.Overview of Cytochrome P4501B1 gene mutations in patients with primary congenital glaucoma.Exp Eye Res 2011;93(5):572-579.
21 Micheal S,Ayub H,Zafar SN,Bakker B,Ali M,Akhtar F,Islam F,Khan MI,Qamar R,den Hollander AI.Identification of novel CYP1B1gene mutations in patients with primary congenital and primary openangle glaucoma.Clin Exp Ophthalmol 2015;43(1):31-39.
22 Firasat S,Riazuddin SA,Khan SN,Riazuddin S.Novel CYP1B1mutations in consanguineous Pakistani families with primary congenital glaucoma.Mol Vis 2008;14:2002-2009.
23 Stoilov I,Akarsu AN,Alozie I,Child A,Barsoum-Homsy M,Turacli ME,Or M,Lewis RA,Ozdemir N,Brice G,Aktan SG,Chevrette L,Coca-Prados M,Sarfarazi M.Sequence analysis and homology modeling suggest that primary congenital glaucoma on 2p21 results from mutations disrupting either the hinge region or the conserved core structures of cytochrome P4501B1.Am J Hum Genet 1998;62(3):573-584.
24 Della Paolera M,de Vasconcellos JP,Umbelino CC,Kasahara N,Rocha MN,Richeti F,Costa VP,Tavares A,de Melo MB.CYP1B1gene analysis in primary congenital glaucoma Brazilian patients:novel mutations and association with poor prognosis.J Glaucoma 2010;19(3):176-182.
25 Medina-Trillo C,Ferre-Fernandez JJ,Aroca-Aguilar JD,BonetFernandez JM,Escribano J.Functional characterization of eight rare missense CYP1B1 variants involved in congenital glaucoma and their association with null genotypes.Acta Ophthalmol 2016;94(7):e555-e560.
26 Su CC,Liu YF,Li SY,Yang JJ,Yen YC.Mutations in the CYP1B1gene may contribute to juvenile-onset open-angle glaucoma.Eye(Lond)2012;26(10):1369-1377.
27 Sena DF,Finzi S,Rodgers K,Del Bono E,Haines JL,Wiggs JL.Founder mutations of CYP1B1 gene in patients with congenital glaucoma from the United States and Brazil.J Med Genet 2004;41(1):e6.
28 Bagiyeva S,Marfany G,Gonzalez-Angulo O,Gonzalez-Duarte R.Mutational screening of CYP1B1 in Turkish PCG families and functional analyses of newly detected mutations.Mol Vis 2007;13:1458-1468.
29 Chakrabarti S,Kaur K,Kaur I,Mandal AK,Parikh RS,Thomas R,Majumder PP.Globally,CYP1B1 mutations in primary congenital glaucoma are strongly structured by geographic and haplotype backgrounds.Invest Ophthalmol Vis Sci 2006;47(1):43-47.
2 Ho CL,Walton DS.Primary congenital glaucoma:2004 update.JPediatr Ophthalmol Strabismus 2004;41(5):271-288;quiz 300-1.
3 Berraho A,Serrou A,Fritez N,El Annas A,Bencherifa F,Gaboun F,Hilal L.Genotype-phenotype correlation in Moroccan patients with primary congenital glaucoma.J Glaucoma 2015;24(4):297-305.
4 Morales-Fernandez L,Martinez-De-la-casa JM,Garcia-Bella J,Mendez C,Saenz-Frances F,Garcia M,Escribano J,Garcia-Feijoo J.Clinical variability of primary congenital glaucoma in a Spanish family with Cyp1b1 gene mutations.J Glaucoma 2015;24(8):630-634.
5 Moore DB,Tomkins O,Ben-Zion I.A review of primary congenital glaucoma in the developing world.Surv Ophthalmol2013;58(3):278-285.
6 Bashir R,Sanai M,Azeem A,Altaf I,Saleem F,Naz S.Contribution of GLC3A locus to primary congenital glaucoma in Pakistani population.Pak J Med Sci 2014;30(6):1341-1345.
7 Bejjani BA,Stockton DW,Lewis RA,Tomey KF,Dueker DK,Jabak M,Astle WF,Lupski JR.Multiple CYP1B1 mutations and incomplete penetrance in an inbred population segregating primary congenital glaucoma suggest frequent de novo events and a dominant modifier locus.Hum Mol Genet 2000;9(3):367-374.
8 Reis LM,Tyler RC,Weh E,Hendee KE,Schilter KF,Phillips JA 3rd,Sequeira S,Schinzel A,Semina EV.Whole exome sequencing identifies multiple diagnoses in congenital glaucoma with systemic anomalies.Clin Genet 2016;90(4):378-382.
9 Souma T,Tompson SW,Thomson BR,et al.Angiopoietin receptor TEKmutations underlie primary congenital glaucoma with variable expressivity.J Clin Invest 2016;126(7):2575-2587.
10 Souzeau E,Hayes M,Ruddle JB,Elder JE,Staffieri SE,Kearns LS,Mackey DA,Zhou T,Ridge B,Burdon KP,Dubowsky A,Craig JE.CYP1B1 copy number variation is not a major contributor to primary congenital glaucoma.Mol Vis 2015;21:160-164.
11 Sheikh SA,Waryah AM,Narsani AK,Shaikh H,Gilal IA,Shah K,Qasim M,Memon AI,Kewalramani P,Shaikh N.Mutational spectrum of the CYP1B1 gene in Pakistani patients with primary congenital glaucoma:novel variants and genotype-phenotype correlations.Mol Vis2014;20:991-1001.
12 Grimberg J,Nawoschik S,Belluscio L,McKee R,Turck A,Eisenberg A.A simple and efficient non-organic procedure for the isolation of genomic DNA from blood.Nucleic Acids Res 1989;17(20):8390.
13 Waryah AM,Narsani AK,Sheikh SA,Shaikh H,Shahani MY.The novel heterozygous Thr377Arg MYOC mutation causes severe Juvenile Open Angle Glaucoma in a large Pakistani family.Gene 2013;528(2):356-359.
14 Ohtake Y,Kubota R,Tanino T,Miyata H,Mashima Y.Novel compound heterozygous mutations in the cytochrome P4501B1 gene(CYP1B1)in a Japanese patient with primary congenital glaucoma.Ophthalmic Genet 2000;21(3):191-193.
15 Chen L,Huang L,Zeng A,He J.CYP1B1 gene mutations with incomplete penetrance in a Chinese pedigree with primary congenital glaucoma:a case report and review of literatures.Int J Clin Exp Med2015;8(8):14538-14541.
16 Stoilov IR,Costa VP,Vasconcellos JP,Melo MB,Betinjane AJ,Carani JC,Oltrogge EV,Sarfarazi M.Molecular genetics of primary congenital glaucoma in Brazil.Invest Ophthalmol Vis Sci 2002;43(6):1820-1827.
17 Rauf B,Irum B,Kabir F,Firasat S,Naeem MA,Khan SN,Husnain T,Riazuddin S,Akram J,Riazuddin SA.A spectrum of CYP1B1 mutations associated with primary congenital glaucoma in families of Pakistani descent.Hum Genome Var 2016;3:16021.
18 Chitsazian F,Tusi BK,Elahi E,et al.CYP1B1 mutation profile of Iranian primary congenital glaucoma patients and associated haplotypes.JMol Diagn 2007;9(3):382-393.
19 Tanwar M,Dada T,Sihota R,Das TK,Yadav U,Dada R.Mutation spectrum of CYP1B1 in North Indian congenital glaucoma patients.Mol Vis 2009;15:1200-1209.
20 Li N,Zhou Y,Du L,Wei M,Chen X.Overview of Cytochrome P4501B1 gene mutations in patients with primary congenital glaucoma.Exp Eye Res 2011;93(5):572-579.
21 Micheal S,Ayub H,Zafar SN,Bakker B,Ali M,Akhtar F,Islam F,Khan MI,Qamar R,den Hollander AI.Identification of novel CYP1B1gene mutations in patients with primary congenital and primary openangle glaucoma.Clin Exp Ophthalmol 2015;43(1):31-39.
22 Firasat S,Riazuddin SA,Khan SN,Riazuddin S.Novel CYP1B1mutations in consanguineous Pakistani families with primary congenital glaucoma.Mol Vis 2008;14:2002-2009.
23 Stoilov I,Akarsu AN,Alozie I,Child A,Barsoum-Homsy M,Turacli ME,Or M,Lewis RA,Ozdemir N,Brice G,Aktan SG,Chevrette L,Coca-Prados M,Sarfarazi M.Sequence analysis and homology modeling suggest that primary congenital glaucoma on 2p21 results from mutations disrupting either the hinge region or the conserved core structures of cytochrome P4501B1.Am J Hum Genet 1998;62(3):573-584.
24 Della Paolera M,de Vasconcellos JP,Umbelino CC,Kasahara N,Rocha MN,Richeti F,Costa VP,Tavares A,de Melo MB.CYP1B1gene analysis in primary congenital glaucoma Brazilian patients:novel mutations and association with poor prognosis.J Glaucoma 2010;19(3):176-182.
25 Medina-Trillo C,Ferre-Fernandez JJ,Aroca-Aguilar JD,BonetFernandez JM,Escribano J.Functional characterization of eight rare missense CYP1B1 variants involved in congenital glaucoma and their association with null genotypes.Acta Ophthalmol 2016;94(7):e555-e560.
26 Su CC,Liu YF,Li SY,Yang JJ,Yen YC.Mutations in the CYP1B1gene may contribute to juvenile-onset open-angle glaucoma.Eye(Lond)2012;26(10):1369-1377.
27 Sena DF,Finzi S,Rodgers K,Del Bono E,Haines JL,Wiggs JL.Founder mutations of CYP1B1 gene in patients with congenital glaucoma from the United States and Brazil.J Med Genet 2004;41(1):e6.
28 Bagiyeva S,Marfany G,Gonzalez-Angulo O,Gonzalez-Duarte R.Mutational screening of CYP1B1 in Turkish PCG families and functional analyses of newly detected mutations.Mol Vis 2007;13:1458-1468.
29 Chakrabarti S,Kaur K,Kaur I,Mandal AK,Parikh RS,Thomas R,Majumder PP.Globally,CYP1B1 mutations in primary congenital glaucoma are strongly structured by geographic and haplotype backgrounds.Invest Ophthalmol Vis Sci 2006;47(1):43-47.