title: Genetics and phenotypes of RPE65 mutations in inherited retinal degeneration.
abstract: OBJECTIVE
  To characterize the spectrum of RPE65 mutations present in 453 patients with retinal dystrophy with an interest in understanding the range of functional deficits attributable to sequence variants in this gene.
  
  
  METHODS
  The 14 exons of RPE65 were amplified by polymerase chain reaction (PCR) from patients' DNA and analyzed for sequence changes by single-strand conformation polymorphism (SSCP) and direct sequencing. Haplotype analysis was performed using RPE65 intragenic polymorphisms. Patients were examined clinically and with visual function tests.
  
  
  RESULTS
  Twenty-one different disease-associated DNA sequence changes predicting missense or nonsense point mutations, insertions, deletions, and splice site defects in RPE65 were identified in 20 patients in homozygous or compound heterozygous form. In one patient, paternal uniparental isodisomy (UPD) of chromosome 1 resulted in homozygosity for a probable functional null allele. Eight of the disease-associated mutations (Y79H, E95Q, E102X, D167Y, 669delCA, IVS7+4a-->g, G436V, and G528V) and one mutation likely to be associated with disease (IVS6+5g-->a) have not been reported previously. The most commonly occurring sequence variant identified in the patients studied was the IVS1+5g-->a mutation, accounting for 9 of 40 (22.5%) total disease alleles. This splice site mutation, as well as R91W, the most common missense mutation, exists on at least two different genetic backgrounds. The phenotype resulting from RPE65 mutations appears to be relatively uniform and independent of mutation class, suggesting that most missense mutations (15 of 40 disease alleles [37.5%]) result in loss of function. At young ages, this group of patients has somewhat better subjective visual capacity than is typically associated with Leber congenital amaurosis (LCA) type I, with a number of patients retaining some useful visual function beyond the second decade of life.
  
  
  CONCLUSIONS
  RPE65 mutations account for a significant percentage (11.4%) of disease alleles in patients with early-onset retinal degeneration. The identification and characterization of patients with RPE65 mutations is likely to represent an important resource for future trials of rational therapies for retinal degeneration.
authors.affiliation: Departments of Ophthalmology and Visual Sciences and. Biological Chemistry, University of Michigan Medical School, Ann Arbor, USA. dathom@umich.edu
authors.forename: D A
authors.initials: DA
authors.lastname: Thompson
authors.forename: P
authors.initials: P
authors.lastname: Gyürüs
authors.forename: L L
authors.initials: LL
authors.lastname: Fleischer
authors.forename: E L
authors.initials: EL
authors.lastname: Bingham
authors.forename: C L
authors.initials: CL
authors.lastname: McHenry
authors.forename: E
authors.initials: E
authors.lastname: Apfelstedt-Sylla
authors.forename: E
authors.initials: E
authors.lastname: Zrenner
authors.forename: B
authors.initials: B
authors.lastname: Lorenz
authors.forename: J E
authors.initials: JE
authors.lastname: Richards
authors.forename: S G
authors.initials: SG
authors.lastname: Jacobson
authors.forename: P A
authors.initials: PA
authors.lastname: Sieving
authors.forename: A
authors.initials: A
authors.lastname: Gal
mesh_terms: D000293:Adolescent; D000328:Adult; D017668:Age of Onset; D002352:Carrier Proteins; D002648:Child; D004252:DNA Mutational Analysis; D004596:Electroretinography; D005136:Eye Proteins / Q000235:genetics*; D006239:Haplotypes; D006801:Humans; D008875:Middle Aged; D020125:Mutation, Missense; D010641:Phenotype; D010857:Pigment Epithelium of Eye / Q000473:pathology*; D016133:Polymerase Chain Reaction; D018807:Polymorphism, Single-Stranded Conformational; D015995:Prevalence; D011506:Proteins / Q000235:genetics*; D012160:Retina / Q000502:physiology; D012162:Retinal Degeneration / Q000453:epidemiology / Q000235:genetics* / Q000473:pathology; D017422:Sequence Analysis, DNA; D014792:Visual Acuity; D019745:cis-trans-Isomerases; D000099178:Retinoid Isomerohydrolase
chemical_list: D002352:Carrier Proteins; D005136:Eye Proteins; D011506:Proteins; D019745:cis-trans-Isomerases; D000099178:Retinoid Isomerohydrolase