Screening of common CYP1B1 mutations in Iranian POAG patients using a microarray-based PrASE protocol
PURPOSE: The gene coding cytochrome P4501B1 (CYP1B1) has been shown to be a major cause of primary congenital glaucoma in the Iranian population. More recently it was shown to also be important in juvenile-onset open angle glaucoma (JOAG). We aimed to further investigate the role of CYP1B1 in a larg...
Autores principales: | , , , , , , , , , , , |
---|---|
Formato: | Texto |
Lenguaje: | English |
Publicado: |
Molecular Vision
2008
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2603445/ https://www.ncbi.nlm.nih.gov/pubmed/19096718 |
_version_ | 1782162590727667712 |
---|---|
author | Suri, Fatemeh Kalhor, Reza Zargar, Seyed Jalal Nilforooshan, Navid Yazdani, Shahin Nezari, Hossein Paylakhi, Seyed Hassan Narooie-Nejhad, Mehrnaz Bayat, Behnaz Sedaghati, Tina Ahmadian, Afshin Elahi, Elahe |
author_facet | Suri, Fatemeh Kalhor, Reza Zargar, Seyed Jalal Nilforooshan, Navid Yazdani, Shahin Nezari, Hossein Paylakhi, Seyed Hassan Narooie-Nejhad, Mehrnaz Bayat, Behnaz Sedaghati, Tina Ahmadian, Afshin Elahi, Elahe |
author_sort | Suri, Fatemeh |
collection | PubMed |
description | PURPOSE: The gene coding cytochrome P4501B1 (CYP1B1) has been shown to be a major cause of primary congenital glaucoma in the Iranian population. More recently it was shown to also be important in juvenile-onset open angle glaucoma (JOAG). We aimed to further investigate the role of CYP1B1 in a larger cohort of primary open angle glaucoma (POAG) patients which included late-onset patients. We also aimed to set up a microarray based protocol for mutation screening with an intent of using the protocol in a future population level screening program. METHODS: Sixty three POAG patients, nine affected family members, and thirty three previously genotyped primary congenital glaucoma (PCG) patients were included in the study. Clinical examination included slit lamp biomicroscopy, IOP measurement, gonioscopic evaluation, fundus examination, and measurement of perimetry. G61E, R368H, R390H, and R469W were screened by a protocol that included multiplexed allele specific amplification in the presence of a protease (PrASE), use of sequence tagged primers, and hybridization to generic arrays on microarray slides. The entire coding sequences of CYP1B1 and myocilin (MYOC) genes were sequenced in all individuals assessed by the microarray assay to carry a mutation. Intragenic single nucleotide polymorphism (SNP) haplotpes were determined for mutated alleles. RESULTS: Genotypes assessed by the array-based PrASE methodology were in 100% concordance with sequencing results. Seven mutation carrying POAG patients (11.1%) were identified, and their distribution was quite skewed between the juvenile-onset individuals (5/21) as compared to late-onset cases (2/42). Four of the seven mutation carrying Iranian patients harbored two mutated alleles. CYP1B1 mutated alleles in Iranian PCG and POAG patients shared common haplotypes. MYOC mutations were not observed in any of the patients. CONCLUSIONS: The PrASE approach allowed reliable simultaneous genotyping of many individuals. It can be an appropriate tool for screening common mutations in large sample sizes. The results suggest that CYP1B1 is implicated in POAG among Iranians, notably in the juvenile-onset form. Contrary to POAG patients studied in other populations, many mutation harboring Iranian patients carry two mutated alleles. We propose an explanation for this observation. |
format | Text |
id | pubmed-2603445 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2008 |
publisher | Molecular Vision |
record_format | MEDLINE/PubMed |
spelling | pubmed-26034452008-12-18 Screening of common CYP1B1 mutations in Iranian POAG patients using a microarray-based PrASE protocol Suri, Fatemeh Kalhor, Reza Zargar, Seyed Jalal Nilforooshan, Navid Yazdani, Shahin Nezari, Hossein Paylakhi, Seyed Hassan Narooie-Nejhad, Mehrnaz Bayat, Behnaz Sedaghati, Tina Ahmadian, Afshin Elahi, Elahe Mol Vis Research Article PURPOSE: The gene coding cytochrome P4501B1 (CYP1B1) has been shown to be a major cause of primary congenital glaucoma in the Iranian population. More recently it was shown to also be important in juvenile-onset open angle glaucoma (JOAG). We aimed to further investigate the role of CYP1B1 in a larger cohort of primary open angle glaucoma (POAG) patients which included late-onset patients. We also aimed to set up a microarray based protocol for mutation screening with an intent of using the protocol in a future population level screening program. METHODS: Sixty three POAG patients, nine affected family members, and thirty three previously genotyped primary congenital glaucoma (PCG) patients were included in the study. Clinical examination included slit lamp biomicroscopy, IOP measurement, gonioscopic evaluation, fundus examination, and measurement of perimetry. G61E, R368H, R390H, and R469W were screened by a protocol that included multiplexed allele specific amplification in the presence of a protease (PrASE), use of sequence tagged primers, and hybridization to generic arrays on microarray slides. The entire coding sequences of CYP1B1 and myocilin (MYOC) genes were sequenced in all individuals assessed by the microarray assay to carry a mutation. Intragenic single nucleotide polymorphism (SNP) haplotpes were determined for mutated alleles. RESULTS: Genotypes assessed by the array-based PrASE methodology were in 100% concordance with sequencing results. Seven mutation carrying POAG patients (11.1%) were identified, and their distribution was quite skewed between the juvenile-onset individuals (5/21) as compared to late-onset cases (2/42). Four of the seven mutation carrying Iranian patients harbored two mutated alleles. CYP1B1 mutated alleles in Iranian PCG and POAG patients shared common haplotypes. MYOC mutations were not observed in any of the patients. CONCLUSIONS: The PrASE approach allowed reliable simultaneous genotyping of many individuals. It can be an appropriate tool for screening common mutations in large sample sizes. The results suggest that CYP1B1 is implicated in POAG among Iranians, notably in the juvenile-onset form. Contrary to POAG patients studied in other populations, many mutation harboring Iranian patients carry two mutated alleles. We propose an explanation for this observation. Molecular Vision 2008-12-18 /pmc/articles/PMC2603445/ /pubmed/19096718 Text en http://creativecommons.org/licenses/by/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Research Article Suri, Fatemeh Kalhor, Reza Zargar, Seyed Jalal Nilforooshan, Navid Yazdani, Shahin Nezari, Hossein Paylakhi, Seyed Hassan Narooie-Nejhad, Mehrnaz Bayat, Behnaz Sedaghati, Tina Ahmadian, Afshin Elahi, Elahe Screening of common CYP1B1 mutations in Iranian POAG patients using a microarray-based PrASE protocol |
title | Screening of common CYP1B1 mutations in Iranian POAG patients using a microarray-based PrASE protocol |
title_full | Screening of common CYP1B1 mutations in Iranian POAG patients using a microarray-based PrASE protocol |
title_fullStr | Screening of common CYP1B1 mutations in Iranian POAG patients using a microarray-based PrASE protocol |
title_full_unstemmed | Screening of common CYP1B1 mutations in Iranian POAG patients using a microarray-based PrASE protocol |
title_short | Screening of common CYP1B1 mutations in Iranian POAG patients using a microarray-based PrASE protocol |
title_sort | screening of common cyp1b1 mutations in iranian poag patients using a microarray-based prase protocol |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2603445/ https://www.ncbi.nlm.nih.gov/pubmed/19096718 |
work_keys_str_mv | AT surifatemeh screeningofcommoncyp1b1mutationsiniranianpoagpatientsusingamicroarraybasedpraseprotocol AT kalhorreza screeningofcommoncyp1b1mutationsiniranianpoagpatientsusingamicroarraybasedpraseprotocol AT zargarseyedjalal screeningofcommoncyp1b1mutationsiniranianpoagpatientsusingamicroarraybasedpraseprotocol AT nilforooshannavid screeningofcommoncyp1b1mutationsiniranianpoagpatientsusingamicroarraybasedpraseprotocol AT yazdanishahin screeningofcommoncyp1b1mutationsiniranianpoagpatientsusingamicroarraybasedpraseprotocol AT nezarihossein screeningofcommoncyp1b1mutationsiniranianpoagpatientsusingamicroarraybasedpraseprotocol AT paylakhiseyedhassan screeningofcommoncyp1b1mutationsiniranianpoagpatientsusingamicroarraybasedpraseprotocol AT narooienejhadmehrnaz screeningofcommoncyp1b1mutationsiniranianpoagpatientsusingamicroarraybasedpraseprotocol AT bayatbehnaz screeningofcommoncyp1b1mutationsiniranianpoagpatientsusingamicroarraybasedpraseprotocol AT sedaghatitina screeningofcommoncyp1b1mutationsiniranianpoagpatientsusingamicroarraybasedpraseprotocol AT ahmadianafshin screeningofcommoncyp1b1mutationsiniranianpoagpatientsusingamicroarraybasedpraseprotocol AT elahielahe screeningofcommoncyp1b1mutationsiniranianpoagpatientsusingamicroarraybasedpraseprotocol |