Investigative Ophthalmology & Visual Science Cover Image for Volume 65, Issue 7
June 2024
Volume 65, Issue 7
Open Access
ARVO Annual Meeting Abstract  |   June 2024
A Comprehensive Bioinformatics Pipeline for Analysing Whole-Genome Sequencing Data of Patients with Inherited Retinal Degenerations
Author Affiliations & Notes
  • Ahmed Shalaby
    University of Oxford Nuffield Department of Clinical Neurosciences, Oxford, Oxfordshire, United Kingdom
  • Samantha R De Silva
    University of Oxford Nuffield Department of Clinical Neurosciences, Oxford, Oxfordshire, United Kingdom
  • Stephanie Halford
    University of Oxford Nuffield Department of Clinical Neurosciences, Oxford, Oxfordshire, United Kingdom
  • Jing Yu
    University of Oxford Nuffield Department of Clinical Neurosciences, Oxford, Oxfordshire, United Kingdom
  • Matteo Ferla
    Department of Statistics, University of Oxford Medical Sciences Division, Oxford, Oxfordshire, United Kingdom
  • Susan M Downes
    University of Oxford Nuffield Department of Clinical Neurosciences, Oxford, Oxfordshire, United Kingdom
  • Footnotes
    Commercial Relationships   Ahmed Shalaby None; Samantha De Silva None; Stephanie Halford None; Jing Yu None; Matteo Ferla None; Susan Downes None
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science June 2024, Vol.65, 3169. doi:
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      Ahmed Shalaby, Samantha R De Silva, Stephanie Halford, Jing Yu, Matteo Ferla, Susan M Downes; A Comprehensive Bioinformatics Pipeline for Analysing Whole-Genome Sequencing Data of Patients with Inherited Retinal Degenerations. Invest. Ophthalmol. Vis. Sci. 2024;65(7):3169.

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

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Abstract

Purpose : To develop a bioinformatic pipeline to increase the diagnostic rate of inherited retinal degenerations (IRDs) by analysing whole-genome sequencing data (WGS) from patients who have not received a molecular diagnosis via panel testing.

Methods : We designed a bioinformatic pipeline to interrogate 732 genes with reported retinal expression. The pipeline was applied to WGS data from 44 patients in whom no pathogenic variants had been identified in known retinal disease genes using panel-based testing. In silico proteomics analysis was performed for all novel variants, variants of uncertain significance and variants in new candidate genes, to filter and select variants for further functional assays.

Results : Potential pathogenic or likely pathogenic variants that could explain the phenotype were identified in 41/44 patients. These included variants in known disease genes not identified by panel testing in 8 patients, novel variants or variants of uncertain significance in previously reported retinal disease genes in 26 patients, and variants in novel genes expressed in the retina in 7 patients. The pipeline did not identify variants for 3 patients.

Conclusions : This comprehensive bioinformatics pipeline, inclusive of in silico proteomic filtering, enabled the identification of candidate pathogenic variants in a high proportion of patients with inherited retinal degenerations lacking a genetic diagnosis after standard panel testing. It also identified new genes that may be associated with retinal disease. Further work is ongoing to confirm the pathogenicity and function of these variants.

This abstract was presented at the 2024 ARVO Annual Meeting, held in Seattle, WA, May 5-9, 2024.

 

Example: fundus imaging of a patient with compound heterozygous mutation in IFT140 gene (p.(His24Tyr) and p.(Arg871His)). (A) Optical coherence tomography (OCT) shows bilateral retinoschisis and loss of photoreceptor outer segment/inner segment (OS/IS). (B) Fundus autofluorescence shows bilateral hypoautofluorescence in the peripapillary, macular, and peripheral retinal regions

Example: fundus imaging of a patient with compound heterozygous mutation in IFT140 gene (p.(His24Tyr) and p.(Arg871His)). (A) Optical coherence tomography (OCT) shows bilateral retinoschisis and loss of photoreceptor outer segment/inner segment (OS/IS). (B) Fundus autofluorescence shows bilateral hypoautofluorescence in the peripapillary, macular, and peripheral retinal regions

 

In-silico analysis of IFT140 mutations (in pink). (A) The His24 residue is in a highly conserved region near the IFT140 N-terminal. (B) The Arg871 residue is predicted to be within the tetratricopeptide repeat (TPR) region. Substitution of mutations at either His24 or Arg871 could disrupt critical protein-protein interactions necessary for the assembly of the intraflagellar transport complex A (IFT-A)

In-silico analysis of IFT140 mutations (in pink). (A) The His24 residue is in a highly conserved region near the IFT140 N-terminal. (B) The Arg871 residue is predicted to be within the tetratricopeptide repeat (TPR) region. Substitution of mutations at either His24 or Arg871 could disrupt critical protein-protein interactions necessary for the assembly of the intraflagellar transport complex A (IFT-A)

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