title: Leu432Val polymorphism in CYP1B1 as a susceptible factor towards predisposition to primary open-angle glaucoma.
abstract: OBJECTIVE
  Defects in cytochrome P450 1B1 (CYP1B1) cause primary congenital glaucoma. However, defects in the gene have also been reported in primary open-angle glaucoma (POAG). Since POAG is primarily a complex disease, we examined the potential of coding single nucleotide polymorphisms (cSNPs) in the gene for association with the disease.
  
  
  METHODS
  Five coding SNPs - c.514 C>G (Arg48Gly), c.727 G>T (Ala119Ser), c.1666 C>G (Leu432Val), c.1719 C>T (Asp449Asp), and c.1730 A>G (Asn453Ser) - were genotyped in 264 unrelated POAG patients and 95 controls. In addition, 542 normal individuals selected from various ethnic groups representing the Indian population were also genotyped for these cSNPs. The patterns of linkage disequilibrium between the SNPs and haplotype variations for comparison between POAG patients and controls as well as different ethnic groups of the Indian population were determined using Haploview. Allelic variants of Leu432Val were cloned by site-directed mutagenesis of normal CYP1B1 cDNA, which were used for transfection of retinal pigment epithelium (RPE) cells. The generation of reactive oxygen species (ROS) was quantified by measuring fluorescence emission by degradation of CM-H2DCFDA using a fluoremeter.
  
  
  RESULTS
  The c.1666G allele of the Leu432Val in CYP1B1 showed a statistically significant higher representation among POAG patients compared to controls (p=0.0001; Odds ratio=6.027; 95% CI: 3.863-9.401) suggesting it to be a potential risk allele toward disease predisposition. Analysis of genotype frequencies of the polymorphism between the two groups demonstrated GG as a potential risk genotype (p=0.0001; Odds ratio=15.505; 95% CI: 5.529-43.474) for the disease. CYP1B1 Val432 was estimated to generate higher ROS in RPE cells compared to its allelic variant (Leu432; p=0.0245 for 15 min and p=0.0197 for 30 min). Comparison of haplotype diversities revealed CGGTA as the risk haplotype for the disease (p=0.0001, by Fisher's exact test).
  
  
  CONCLUSIONS
  We report CYP1B1 c.1666G (Val432) as a susceptible allele for POAG and CGGTA as the risk haplotype for the disease. Higher ROS generation by Val432 in CYP1B1 might lead to apoptotic change that leads to glaucoma. Remarkable variation of the cSNPs observed among ethnic groups of India could provide insight for future epidemiological studies on POAG in these population groups.
authors.affiliation: Molecular and Human Genetics Division, Indian Institute of Chemical Biology (Council of Scientific & Industrial Research), Kolkata, India.
authors.forename: Ashima
authors.initials: A
authors.lastname: Bhattacharjee
authors.forename: Deblina
authors.initials: D
authors.lastname: Banerjee
authors.forename: Suddhasil
authors.initials: S
authors.lastname: Mookherjee
authors.forename: Moulinath
authors.initials: M
authors.lastname: Acharya
authors.forename: Antara
authors.initials: A
authors.lastname: Banerjee
authors.forename: Ananya
authors.initials: A
authors.lastname: Ray
authors.forename: Abhijit
authors.initials: A
authors.lastname: Sen
authors.forename: Kunal
authors.initials: K
authors.lastname: Ray
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001189:Aryl Hydrocarbon Hydroxylases; D044466:Asian People / Q000235:genetics; D016022:Case-Control Studies; D002460:Cell Line; D002648:Child; D065633:Cytochrome P-450 CYP1B1; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics*; D005006:Ethnicity / Q000235:genetics; D005787:Gene Frequency; D020022:Genetic Predisposition to Disease; D005843:Geography; D005902:Glaucoma, Open-Angle / Q000201:enzymology* / Q000235:genetics*; D006239:Haplotypes; D006801:Humans; D007194:India; D007930:Leucine / Q000235:genetics*; D015810:Linkage Disequilibrium / Q000235:genetics; D008875:Middle Aged; D010857:Pigment Epithelium of Eye / Q000166:cytology / Q000378:metabolism; D020641:Polymorphism, Single Nucleotide / Q000235:genetics*; D017382:Reactive Oxygen Species / Q000378:metabolism; D012984:Software; D014633:Valine / Q000235:genetics*
chemical_list: D017382:Reactive Oxygen Species; D003577:Cytochrome P-450 Enzyme System; D001189:Aryl Hydrocarbon Hydroxylases; C585496:CYP1B1 protein, human; D065633:Cytochrome P-450 CYP1B1; D007930:Leucine; D014633:Valine