June 2023
Volume 64, Issue 8
Open Access
ARVO Annual Meeting Abstract  |   June 2023
Finding the Second Variant: a Purpose-built Bioinformatics Pipeline
Author Affiliations & Notes
  • Ahmed Shalaby
    Nuffield Department of Clinical Neurosciences, University of Oxford, Oxford, Oxfordshire, United Kingdom
  • Samantha R De Silva
    Nuffield Department of Clinical Neurosciences, University of Oxford, Oxford, Oxfordshire, United Kingdom
  • Jing Yu
    Nuffield Department of Clinical Neurosciences, University of Oxford, Oxford, Oxfordshire, United Kingdom
  • Stephanie Halford
    Nuffield Department of Clinical Neurosciences, University of Oxford, Oxford, Oxfordshire, United Kingdom
  • Susan M Downes
    Nuffield Department of Clinical Neurosciences, University of Oxford, Oxford, Oxfordshire, United Kingdom
  • Footnotes
    Commercial Relationships   Ahmed Shalaby None; Samantha De Silva None; Jing Yu None; Stephanie Halford None; Susan Downes None
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science June 2023, Vol.64, 4503. doi:
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    • Get Citation

      Ahmed Shalaby, Samantha R De Silva, Jing Yu, Stephanie Halford, Susan M Downes; Finding the Second Variant: a Purpose-built Bioinformatics Pipeline. Invest. Ophthalmol. Vis. Sci. 2023;64(8):4503.

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

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Abstract

Purpose : To date, approximately 60% of inherited retinal degenerations patients still do not have a molecular diagnosis despite genetic testing. In this study, we aimed to design a bioinformatic pipeline to identify the second variant in patients in whom only a single variant had previously been identified in a presumed autosomal recessively inherited gene consistent with the clinical phenotype

Methods : We included 27 patients with IRDs recruited to the 100,000 Genomes Project (GEL) from the Oxford Eye Hospital in whom whole-genome sequencing (WGS) was performed for the proband and 2 family members where possible.
A bioinformatics workflow was designed encompassing a wide range of publicly available bioinformatics tools such as PolyPhen-2, SIFT, CADD, and UTRannotator in addition to a custom-made tool designed by one of the authors called SVrare. The latter analyses and prioritises structural variants identified by CANVAS and MANTA

Results : Of the 27 patients included, 9 were solved at different stages of the pipeline. The novel variants varied between structural variants, intronic variants that affect splicing, and single nucleotide variants in coding regions that skipped the conventional bioinformatics pipeline as they either had low pathogenicity scores or were considered too common to be pathogenic. In this study, we present the phenotype of the solved patients and their genotype as identified at each stage of the purpose-built pipeline

Conclusions : We demonstrate a substantial increase in the rate of confirming a genetic diagnosis in patients with IRDs using a bioinformatic pipeline to perform a comprehensive analysis of candidate genes. This work also shows the added value of scrutiny of whole-genome sequencing data of undiagnosed patients by a research team working within a multidisciplinary framework

This abstract was presented at the 2023 ARVO Annual Meeting, held in New Orleans, LA, April 23-27, 2023.

 

Figure 1: Summary of the steps in the workflow used in this study.

Figure 1: Summary of the steps in the workflow used in this study.

 

Example case of a patient who was diagnosed at the 2nd step of the pipeline (relaxed filters analysis). He was diagnosed with oculocutaneous albinism at the age of 1 year old. Only one variant in TYR gene was identified p.(Thr373Lys). Relaxed filters analysis identified two other common variants in TYR: (p.(Ser192Tyr), CADD score= 23.8 and gnomAD AF= 0.33) and (p.(Arg402Gln), CADD score= 27.2, gnomAD AF= 0.18). This is consistent with the rare occurrence of the triallelic inheritance pattern of oculocutaneous albinism that has been reported before in a UK cohort

Example case of a patient who was diagnosed at the 2nd step of the pipeline (relaxed filters analysis). He was diagnosed with oculocutaneous albinism at the age of 1 year old. Only one variant in TYR gene was identified p.(Thr373Lys). Relaxed filters analysis identified two other common variants in TYR: (p.(Ser192Tyr), CADD score= 23.8 and gnomAD AF= 0.33) and (p.(Arg402Gln), CADD score= 27.2, gnomAD AF= 0.18). This is consistent with the rare occurrence of the triallelic inheritance pattern of oculocutaneous albinism that has been reported before in a UK cohort

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