Predicting the pathogenicity of RPE65 mutations.

Journal Article (Journal Article)

To assist in distinguishing disease-causing mutations from nonpathogenic polymorphisms, we developed an objective algorithm to calculate an "estimate of pathogenic probability" (EPP) based on the prevalence of a specific variation, its segregation within families, and its predicted effects on protein structure. Eleven missense variations in the RPE65 gene were evaluated in patients with Leber congenital amaurosis (LCA) using the EPP algorithm. The accuracy of the EPP algorithm was evaluated using a cell-culture assay of RPE65-isomerase activity The variations were engineered into plasmids containing a human RPE65 cDNA and the retinoid isomerase activity of each variant was determined in cultured cells. The EPP algorithm predicted eight substitution mutations to be disease-causing variants. The isomerase catalytic activities of these RPE65 variants were all less than 6% of wild-type. In contrast, the EPP algorithm predicted the other three substitutions to be non-disease-causing, with isomerase activities of 68%, 127%, and 110% of wild-type, respectively. We observed complete concordance between the predicted pathogenicities of missense variations in the RPE65 gene and retinoid isomerase activities measured in a functional assay. These results suggest that the EPP algorithm may be useful to evaluate the pathogenicity of missense variations in other disease genes where functional assays are not available.

Full Text

Duke Authors

Cited Authors

  • Philp, AR; Jin, M; Li, S; Schindler, EI; Iannaccone, A; Lam, BL; Weleber, RG; Fishman, GA; Jacobson, SG; Mullins, RF; Travis, GH; Stone, EM

Published Date

  • August 2009

Published In

Volume / Issue

  • 30 / 8

Start / End Page

  • 1183 - 1188

PubMed ID

  • 19431183

Pubmed Central ID

  • PMC2717180

Electronic International Standard Serial Number (EISSN)

  • 1098-1004

Digital Object Identifier (DOI)

  • 10.1002/humu.21033


  • eng

Conference Location

  • United States