title: CYP1B1, MYOC, and LTBP2 mutations in primary congenital glaucoma patients in the United States.
abstract: OBJECTIVE
  To screen primary congenital glaucoma patients in the United States for sequence variants within the CYP1B1, LTBP2, and MYOC genes using Sanger and whole exome sequencing.
  
  
  METHODS
  Retrospective case-control study.
  
  
  METHODS
  Fifty-seven primary congenital glaucoma patients (47 families), 71 unaffected family members of the primary congenital glaucoma probands, and 101 healthy unrelated individuals were recruited from a single institution. Sanger sequencing of the primary congenital glaucoma gene, CYP1B1, was performed on 47 proband deoxyribonucleic acid samples. Simultaneously, whole exome sequencing was conducted on 3 families, each including more than 1 affected individual. Concurrently, 33 of 47 primary congenital glaucoma probands with extended family deoxyribonucleic acid samples were screened for LTBP2 and MYOC gene mutations. Exome-sequenced variations were validated by additional Sanger sequencing to confirm segregation of filtered disease-causing single nucleotide variations.
  
  
  RESULTS
  Seven primary congenital glaucoma families (14.9%) manifested disease phenotypes attributable to CYP1B1 mutations. One primary congenital glaucoma family possessed homozygous mutant alleles, whereas 6 families carried compound heterozygous mutations. Five novel combinations of compound heterozygous mutations were identified, of which 2 combinations were found with whole exome sequencing. No disease-causing mutations within the LTBP2 and MYOC genes were discovered.
  
  
  CONCLUSIONS
  This study analyzed CYP1B1, LTBP2, and MYOC mutations in a cohort of primary congenital glaucoma patients from the United States, applying whole exome sequencing as a complementary tool to Sanger sequencing. Whole exome sequencing, coupled with Sanger sequencing, may identify novel genes in primary congenital glaucoma patients who have no mutations in known primary congenital glaucoma genes.
authors.affiliation: Duke-National University of Singapore Graduate Medical School, Singapore, Republic of Singapore.
authors.forename: Sing-Hui
authors.initials: SH
authors.lastname: Lim
authors.forename: Khanh-Nhat
authors.initials: KN
authors.lastname: Tran-Viet
authors.forename: Tammy L
authors.initials: TL
authors.lastname: Yanovitch
authors.forename: Sharon F
authors.initials: SF
authors.lastname: Freedman
authors.forename: Thomas
authors.initials: T
authors.lastname: Klemm
authors.forename: Whitney
authors.initials: W
authors.lastname: Call
authors.forename: Caldwell
authors.initials: C
authors.lastname: Powell
authors.forename: Ajay
authors.initials: A
authors.lastname: Ravichandran
authors.forename: Ravikanth
authors.initials: R
authors.lastname: Metlapally
authors.forename: Erica B
authors.initials: EB
authors.lastname: Nading
authors.forename: Steve
authors.initials: S
authors.lastname: Rozen
authors.forename: Terri L
authors.initials: TL
authors.lastname: Young
mesh_terms: D001189:Aryl Hydrocarbon Hydroxylases / Q000235:genetics*; D016022:Case-Control Studies; D065633:Cytochrome P-450 CYP1B1; D003598:Cytoskeletal Proteins / Q000235:genetics*; D004252:DNA Mutational Analysis; D059472:Exome / Q000235:genetics; D005136:Eye Proteins / Q000235:genetics*; D005260:Female; D006023:Glycoproteins / Q000235:genetics*; D006801:Humans; D006871:Hydrophthalmos / Q000208:ethnology / Q000235:genetics*; D007429:Intraocular Pressure; D053779:Latent TGF-beta Binding Proteins / Q000235:genetics*; D008297:Male; D009154:Mutation; D010375:Pedigree; D016133:Polymerase Chain Reaction; D020641:Polymorphism, Single Nucleotide; D012189:Retrospective Studies; D014481:United States / Q000453:epidemiology; D000099179:Myocilin
chemical_list: D001189:Aryl Hydrocarbon Hydroxylases; D065633:Cytochrome P-450 CYP1B1; D003598:Cytoskeletal Proteins; D005136:Eye Proteins; D006023:Glycoproteins; D053779:Latent TGF-beta Binding Proteins; D000099179:Myocilin; C509035:LTBP2 protein, human; C585496:CYP1B1 protein, human