Assessing the pathogenicity of RYR1 variants in malignant hyperthermia

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Merritt, A ; Booms, P ; Shaw, M-A ; Miller, DM ; Daly, C ; Bilmen, JG ; Stowell, KM ; Allen, PD ; Steele, DS ; Hopkins, PM (2017)
  • Publisher: Oxford University Press

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.
  • References (34)
    34 references, page 1 of 4

    1. Hopkins PM. Malignant hyperthermia Ð pharmacology of triggering. Br J Anaesth 2011; 107:48-56

    2. Weiss RG, OÕConnell KM, Flucher BE, Allen PD, Grabner M, Dirksen RT. Functional analysis of the R1086H malignant hyperthermia mutation in the DHPR reveals an unexpected influence of the III-IV loop on skeletal muscle EC coupling. Am J Physiol Cell Physiol 2004; 287: C1094-102

    3. Carpenter D, Ringrose C, Leo V, et al. The role of CACNA1S in predisposition to malignant hyperthermia. BMC Med Genet 2009; 10:104

    4. Eltit JM, Bannister RA, Moua O, et al. Malignant hyperthermia susceptibility arising from altered resting coupling between the skeletal muscle L-type Ca2+ channel and the type 1 ryanodine receptor. Proc Natl Acad Sci U S A 2012; 109: 7923-8

    5. Horstick EJ, Linsley JW, Dowling JJ, et al. Stac3 is a component of the excitationcontraction coupling machinery and mutated in Native American myopathy. Nat Commun 2013;4:1952

    6. Schiemann AH, DŸrholt EM, Pollock N, Stowell KM. Sequence capture and massively parallel sequencing to detect mutations associated with malignant hyperthermia. Br J Anaesth 2013;110:122-7

    7. Kim J, Jarvik GP, Browning BL, et al. Exome sequencing reveals novel rare variants in ryanodine receptor and calcium channel genes in malignant hyperthermia families. Anesthesiology 2013;119:1054-65

    8. Gonsalves SG, Ng D, Johnston JJ, et al. Using exome data to identify malignant hyperthermia susceptibility mutations. Anesthesiology 2013;119:1043Ð53

    9. Fiszer D, Shaw M-A, Fisher NA, et al. Next generation sequencing of RYR1 and CACNA1S in malignant hyperthermia and exertional heat illness. Anesthesiology 2015: 122;1033-46

    10. Hopkins PM, RŸffert H, Snoeck MM, et al. The European Malignant Hyperthermia Group guidelines for the investigation of malignant hyperthermia susceptibility. Br J Anaesth 2015; 115: 531-9

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