June 2021
Volume 62, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2021
Numerous Protein Mutations Detected in FEVR Patients Using a New Custom Ampliseq Targeted-Sequencing Panel
Author Affiliations & Notes
  • Amanda Petrelli Cicerone
    Eye Research Institute, Oakland University, Rochester, Michigan, United States
  • Wendy A Dailey
    Eye Research Institute, Oakland University, Rochester, Michigan, United States
  • Michael Sun
    Eye Research Institute, Oakland University, Rochester, Michigan, United States
  • Naomi Haque
    Eye Research Institute, Oakland University, Rochester, Michigan, United States
  • Alvaro Guzman
    Eye Research Institute, Oakland University, Rochester, Michigan, United States
  • Kendra Mellert
    Eye Research Institute, Oakland University, Rochester, Michigan, United States
  • Kimberly A Drenser
    Eye Research Institute, Oakland University, Rochester, Michigan, United States
  • Kenneth P Mitton
    Eye Research Institute, Oakland University, Rochester, Michigan, United States
  • Footnotes
    Commercial Relationships   Amanda Petrelli Cicerone, None; Wendy Dailey, None; Michael Sun, None; Naomi Haque, None; Alvaro Guzman, None; Kendra Mellert, None; Kimberly Drenser, None; Kenneth Mitton, None
  • Footnotes
    Support  Pediatric Retinal Research Foundation
Investigative Ophthalmology & Visual Science June 2021, Vol.62, 1573. doi:
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      Amanda Petrelli Cicerone, Wendy A Dailey, Michael Sun, Naomi Haque, Alvaro Guzman, Kendra Mellert, Kimberly A Drenser, Kenneth P Mitton; Numerous Protein Mutations Detected in FEVR Patients Using a New Custom Ampliseq Targeted-Sequencing Panel. Invest. Ophthalmol. Vis. Sci. 2021;62(8):1573.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : Analyze the first subset of 69 patients with Familial Exudative Vitreo Retinopathy (FEVR) who were sequenced using a custom Ampliseq targeted gene panel that includes 7 genes (NDP, CTNNB1, TSPAN12, KIF11, FZD4, LRP5, ZNF408) that are potentially involved in FEVR.

Methods : A custom Ampliseq targeted-panel (180 amplicons) for 8 genes was designed with illumina’s DesignStudio Sequencing Assay Design for complete coverage of 83 exons with 25 bp adjacent intron sequence. Seven of the genes are applicable to FEVR: NDP (ChrX), CTNNB1 (Chr3); TSPAN12 (Chr7); KIF11 (Chr10), FZD4 (Chr11), LRP5 (Chr11), ZNF408 (Chr11). Ampliseq libraries were pooled and sequenced using the Illumina iSeq-100 platform. Variant impacts and allele frequency data were determined from ClinVar and The Genome Aggregation Databases (gnomAD).

Results : For the seven genes that are applicable to FEVR, a total of 35 variants were found that alter protein sequence, with the following relative distribution: NDP 1/35 (2.9%), CTNNB1 1/35 (2.9%),TSPAN12 1/35 (2.9%), KIF11 3/35 (8.6%), FZD4 10/35 (28.6%), LRP5 13/35 (37.1%), ZNF408 6/35 (17.1%). Types of protein changes were: single amino acid change (77.1%), amino acid deletion (8.6 %), and amino acid insertion (2.9%).

Conclusions : A custom Ampliseq targeted-sequencing Orphan Pediatric Retinal Disease panel (version 2, 8 genes) developed by our lab provided extensive sequencing coverage of all exons and detected 35 protein-altering variants in a set of 69 FEVR patients. 76% of the variants were found in three genes: FZD4, LRP5, and ZNF408. The testing format greatly reduces associated costs and now facilitates greater access to genetic testing for Families with this very rare inherited retinal disease.

This is a 2021 ARVO Annual Meeting abstract.

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