Merritt, A, Booms, P, Shaw, M-A orcid.org/0000-0001-6718-5423 et al. (7 more authors) (2017) Assessing the pathogenicity of RYR1 variants in malignant hyperthermia. British Journal of Anaesthesia, 118 (4). pp. 533-543. ISSN 0007-0912
Abstract
Background: Missense variants in the ryanodine receptor 1 gene (RYR1) are associated with malignant hyperthermia but only a minority of these have met criteria for use in predictive DNA diagnosis. We examined the utility of a simplified method of segregation analysis and a functional assay for determining the pathogenicity of recurrent RYR1 variants associated with malignant hyperthermia. Methods: We identified previously uncharacterised RYR1 variants found in 4 or more malignant hyperthermia families and conducted simplified segregation analyses. An efficient cloning and mutagenesis strategy was used to express ryanodine receptor protein containing one of six RYR1 variants in HEK293 cells. Caffeine-induced calcium release, measured using a fluorescent calcium indicator, was compared in cells expressing each variant to that in cells expressing wild type ryanodine receptor protein. Results: We identified 43 malignant hyperthermia families carrying one of the six RYR1 variants. There was segregation of genotype with the malignant hyperthermia susceptibility phenotype in families carrying the p.E3104K and p.D3986E variants but the number of informative meioses limited the statistical significance of the associations. HEK293 functional assays demonstrated an increased sensitivity of RyR1 channels containing the p.R2336H, p.R2355W, p.E3104K, p.G3990V and p.V4849I compared to wild type but cells expressing p.D3986E had a similar caffeine sensitivity to cells expressing wild type RyR1. Conclusions: Segregation analysis is of limited value in assessing pathogenicity of RYR1 variants in malignant hyperthermia. Functional analyses in HEK293 cells provided evidence to support the use of p.R2336H, p.R2355W, p.E3104K, p.G3990V and p.V4849I for diagnostic purposes but not p.D3986E.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | © The Author 2017. Published by Oxford University Press on behalf of the British Journal of Anaesthesia. All rights reserved. This is a pre-copyedited, author-produced PDF of an article accepted for publication in British Journal of Anaesthesia following peer review. The version of record Merritt, A, Booms, P, Shaw, M-A et al. (7 more authors) (2017) Assessing the pathogenicity of RYR1 variants in malignant hyperthermia. British Journal of Anaesthesia, 118 (4). pp. 533-543. ISSN 0007-0912 is available online at: https://doi.org/10.1093/bja/aex042. Uploaded in accordance with the publisher's self-archiving policy. |
Keywords: | Malignant hyperthermia; genetics, diagnosis, pathophysiology; HEK293; calcium regulation; RYR1 |
Dates: |
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Institution: | The University of Leeds |
Academic Units: | The University of Leeds > Faculty of Biological Sciences (Leeds) > School of Biomedical Sciences (Leeds) The University of Leeds > Faculty of Medicine and Health (Leeds) > School of Medicine (Leeds) > Inst of Biomed & Clin Sciences (LIBACS) (Leeds) > Trans Anaesthetics & Surgical Sciences (Leeds) |
Funding Information: | Funder Grant number British Journal of Anaesthesia NONE GIVEN National Institute of Health - DELETED Not Known |
Depositing User: | Symplectic Publications |
Date Deposited: | 17 Jan 2017 15:35 |
Last Modified: | 08 Apr 2018 00:38 |
Status: | Published |
Publisher: | Oxford University Press |
Identification Number: | 10.1093/bja/aex042 |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:110663 |