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A systematic analysis of splicing variants identifies new diagnoses in the 100,000 Genomes Project

A systematic analysis of splicing variants identifies new diagnoses in the 100,000 Genomes Project
A systematic analysis of splicing variants identifies new diagnoses in the 100,000 Genomes Project
Abstract Background Genomic variants which disrupt splicing are a major cause of rare genetic diseases. However, variants which lie outside of the canonical splice sites are difficult to interpret clinically. Improving the clinical interpretation of non-canonical splicing variants offers a major opportunity to uplift diagnostic yields from whole genome sequencing data. Methods Here, we examine the landscape of splicing variants in whole-genome sequencing data from 38,688 individuals in the 100,000 Genomes Project and assess the contribution of non-canonical splicing variants to rare genetic diseases. We use a variant-level constraint metric (the mutability-adjusted proportion of singletons) to identify constrained functional variant classes near exon–intron junctions and at putative splicing branchpoints. To identify new diagnoses for individuals with unsolved rare diseases in the 100,000 Genomes Project, we identified individuals with de novo single-nucleotide variants near exon–intron boundaries and at putative splicing branchpoints in known disease genes. We identified candidate diagnostic variants through manual phenotype matching and confirmed new molecular diagnoses through clinical variant interpretation and functional RNA studies. Results We show that near-splice positions and splicing branchpoints are highly constrained by purifying selection and harbour potentially damaging non-coding variants which are amenable to systematic analysis in sequencing data. From 258 de novo splicing variants in known rare disease genes, we identify 35 new likely diagnoses in probands with an unsolved rare disease. To date, we have confirmed a new diagnosis for six individuals, including four in whom RNA studies were performed. Conclusions Overall, we demonstrate the clinical value of examining non-canonical splicing variants in individuals with unsolved rare diseases.
figshare
Blakes, Alexander J. M.
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Baralle, Diana
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Douglas, Andrew G. L.
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Lees, Melissa
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Bunyan, David
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Thomas, N. Simon
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Davies, Ian
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Lord, Jenny
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Whiffin, Nicola
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Pichini, Amanda
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O’Donovan, Peter
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Ruiz, April
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Taylor Tavares, Ana Lisa
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Greenhalgh, Lynn
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Thomas, Tessy
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Smithson, Sarah F.
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Burren, Christine P.
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Moledina, Hassan E.
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Wai, Htoo A.
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Blakes, Alexander J. M.
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Baralle, Diana
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Douglas, Andrew G. L.
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Lees, Melissa
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Bunyan, David
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Thomas, N. Simon
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Davies, Ian
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Lord, Jenny
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Whiffin, Nicola
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Pichini, Amanda
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O’Donovan, Peter
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Ruiz, April
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Taylor Tavares, Ana Lisa
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Greenhalgh, Lynn
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Thomas, Tessy
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Smithson, Sarah F.
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Burren, Christine P.
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Moledina, Hassan E.
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Wai, Htoo A.
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(2022) A systematic analysis of splicing variants identifies new diagnoses in the 100,000 Genomes Project. figshare doi:10.6084/m9.figshare.c.6119088 [Dataset]

Record type: Dataset

Abstract

Abstract Background Genomic variants which disrupt splicing are a major cause of rare genetic diseases. However, variants which lie outside of the canonical splice sites are difficult to interpret clinically. Improving the clinical interpretation of non-canonical splicing variants offers a major opportunity to uplift diagnostic yields from whole genome sequencing data. Methods Here, we examine the landscape of splicing variants in whole-genome sequencing data from 38,688 individuals in the 100,000 Genomes Project and assess the contribution of non-canonical splicing variants to rare genetic diseases. We use a variant-level constraint metric (the mutability-adjusted proportion of singletons) to identify constrained functional variant classes near exon–intron junctions and at putative splicing branchpoints. To identify new diagnoses for individuals with unsolved rare diseases in the 100,000 Genomes Project, we identified individuals with de novo single-nucleotide variants near exon–intron boundaries and at putative splicing branchpoints in known disease genes. We identified candidate diagnostic variants through manual phenotype matching and confirmed new molecular diagnoses through clinical variant interpretation and functional RNA studies. Results We show that near-splice positions and splicing branchpoints are highly constrained by purifying selection and harbour potentially damaging non-coding variants which are amenable to systematic analysis in sequencing data. From 258 de novo splicing variants in known rare disease genes, we identify 35 new likely diagnoses in probands with an unsolved rare disease. To date, we have confirmed a new diagnosis for six individuals, including four in whom RNA studies were performed. Conclusions Overall, we demonstrate the clinical value of examining non-canonical splicing variants in individuals with unsolved rare diseases.

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Published date: 1 January 2022

Identifiers

Local EPrints ID: 477731
URI: http://eprints.soton.ac.uk/id/eprint/477731
PURE UUID: 90983a6e-c151-415f-ab16-1f27c9ce1cd2
ORCID for Diana Baralle: ORCID iD orcid.org/0000-0003-3217-4833
ORCID for Andrew G. L. Douglas: ORCID iD orcid.org/0000-0001-5154-6714
ORCID for Jenny Lord: ORCID iD orcid.org/0000-0002-0539-9343
ORCID for Htoo A. Wai: ORCID iD orcid.org/0000-0002-3560-6980

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Date deposited: 13 Jun 2023 17:24
Last modified: 24 Jan 2024 02:55

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Contributors

Contributor: Alexander J. M. Blakes
Contributor: Diana Baralle ORCID iD
Contributor: Andrew G. L. Douglas ORCID iD
Contributor: Melissa Lees
Contributor: David Bunyan
Contributor: N. Simon Thomas
Contributor: Ian Davies
Contributor: Jenny Lord ORCID iD
Contributor: Nicola Whiffin
Contributor: Amanda Pichini
Contributor: Peter O’Donovan
Contributor: April Ruiz
Contributor: Ana Lisa Taylor Tavares
Contributor: Lynn Greenhalgh
Contributor: Tessy Thomas
Contributor: Sarah F. Smithson
Contributor: Christine P. Burren
Contributor: Hassan E. Moledina
Contributor: Htoo A. Wai ORCID iD

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