Identification and Biochemical Characterization of a Novel Ryanodine Receptor Gene Mutation Associated With Malignant Hyperthermia

Background Mutations in the skeletal muscle ryanodine receptor gene may result in altered calcium release from sarcoplasmic reticulum stores, giving rise to malignant hyperthermia (MH). MH is a pharmacogenetic skeletal muscle disorder triggered by volatile anesthetics and depolarizing muscle relaxants. Diagnosis of MH is by in vitro contracture testing of quadriceps muscle. DNA analysis of causative mutations is limited by the large number of mutations that cosegregate with MH and the relatively few that have been biochemically characterized. Methods DNA sequence analysis was used to screen the skeletal muscle ryanodine receptor gene in MH-susceptible individuals. A diagnostic test using real-time polymerase chain reaction was developed to detect the mutation in individuals diagnosed as MH susceptible by in vitro contracture testing. The functional relevance of this mutation was examined in Epstein-Barr virus-immortalized B-lymphoblastoid cells. Results A novel ryanodine receptor mutation (cytosine 14997 thymine resulting in a histidine 4833 tyrosine substitution) was identified in pathology specimens from two patients with fatal MH reactions. B lymphocytes from patients with this mutation were approximately twofold more sensitive than MH-negative cells to activation with 4-chloro-m-cresol. The amount of Ca released from B lymphocytes of MH-susceptible patients was significantly greater than that released from cells of family members without this mutation. Haplotype analysis suggests that both families had a common ancestor. Conclusions DNA analysis to detect mutations which cosegregate with MH as well as biochemical assays on cultured lymphocytes obtained from blood can serve as useful diagnostic tools for MH susceptibility and genotype-phenotype correlations.

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Genetics of Malignant Hyperthermia

The Scientific World JOURNAL ◽

10.1100/tsw.2006.289 ◽

2006 ◽

Vol 6 ◽

pp. 1722-1730 ◽

Cited By ~ 23

Author(s):

Barbara W. Brandom

Keyword(s):

Malignant Hyperthermia ◽

Candidate Genes ◽

Ryanodine Receptor ◽

Receptor Gene ◽

Linkage Studies ◽

Chromosome 19

Study of the genetics of the malignant hyperthermia syndrome began in families in which both malignant hyperthermia (MH) episodes had been experienced and individuals had strongly positive contracture tests diagnostic of susceptibility to MH. Linkage studies associated this MH phenotype to the ryanodine receptor gene (RYR1) at chromosome 19q13.1 in many families. Although the MH phenotype is not always linked to chromosome 19, theRYR1has remained the focus of experimentation. Other candidate genes exist, but few MH-susceptible families have variants of these genes. Hundreds of MH-susceptible people have variants ofRYR1.

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Functional Characterization of a Distinct Ryanodine Receptor Mutation in Human Malignant Hyperthermia-susceptible Muscle

Journal of Biological Chemistry ◽

10.1074/jbc.272.8.5256 ◽

1997 ◽

Vol 272 (8) ◽

pp. 5256-5260 ◽

Cited By ~ 58

Author(s):

Michael Richter ◽

Lothar Schleithoff ◽

Thomas Deufel ◽

Frank Lehmann-Horn ◽

Annegret Herrmann-Frank

Keyword(s):

Malignant Hyperthermia ◽

Ryanodine Receptor ◽

Functional Characterization ◽

Malignant Hyperthermia Susceptible

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Additional evidence that the ryanodine receptor gene (RYR1) causes malignant hyperthermia and severe skeletal malformations

American Journal of Medical Genetics Part A ◽

10.1002/ajmg.a.35678 ◽

2012 ◽

Vol 161 (1) ◽

pp. 234-235

Author(s):

Yosuke Kakisaka ◽

Kazuhiro Haginoya ◽

Yuko Takahashi ◽

Tatsuhiro Ochiai ◽

Ikuma Fujiwara ◽

...

Keyword(s):

Malignant Hyperthermia ◽

Ryanodine Receptor ◽

Receptor Gene ◽

Skeletal Malformations

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Molecular Genetic Analysis of the Ryanodine Receptor Gene (RYR1) in Korean Malignant Hyperthermia Families

Annals of Laboratory Medicine ◽

10.3343/kjlm.2010.30.6.702 ◽

2010 ◽

Vol 30 (6) ◽

pp. 702-710 ◽

Cited By ~ 4

Author(s):

Ho Lee ◽

Dong Chan Kim ◽

Jae Hyeon Lee ◽

Yong Gon Cho ◽

Hye Soo Lee ◽

...

Keyword(s):

Genetic Analysis ◽

Malignant Hyperthermia ◽

Ryanodine Receptor ◽

Receptor Gene ◽

Molecular Genetic ◽

Molecular Genetic Analysis

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Functional properties of ryanodine receptors carrying three amino acid substitutions identified in patients affected by multi-minicore disease and central core disease, expressed in immortalized lymphocytes

Biochemical Journal ◽

10.1042/bj20051282 ◽

2006 ◽

Vol 395 (2) ◽

pp. 259-266 ◽

Cited By ~ 43

Author(s):

Sylvie Ducreux ◽

Francesco Zorzato ◽

Ana Ferreiro ◽

Heinz Jungbluth ◽

Francesco Muntoni ◽

...

Keyword(s):

Amino Acid ◽

Malignant Hyperthermia ◽

Ryanodine Receptor ◽

Receptor Gene ◽

Receptor Activation ◽

Central Core ◽

Central Core Disease ◽

Amino Acid Substitutions ◽

Time Data ◽

Minicore Disease

More than 80 mutations in the skeletal muscle ryanodine receptor gene have been found to be associated with autosomal dominant forms of malignant hyperthermia and central core disease, and with recessive forms of multi-minicore disease. Studies on the functional effects of pathogenic dominant mutations have shown that they mostly affect intracellular Ca2+ homoeostasis, either by rendering the channel hypersensitive to activation (malignant hyperthermia) or by altering the amount of Ca2+ released subsequent to physiological or pharmacological activation (central core disease). In the present paper, we show, for the first time, data on the functional effect of two recently identified recessive ryanodine receptor 1 amino acid substitutions, P3527S and V4849I, as well as that of R999H, another substitution that was identified in two siblings that were affected by multi-minicore disease. We studied the intracellular Ca2+ homoeostasis of EBV (Epstein–Barr virus)-transformed lymphoblastoid cells from the affected patients, their healthy relatives and control individuals. Our results show that the P3527S substitution in the homozygous state affected the amount of Ca2+ released after pharmacological activation with 4-chloro-m-cresol and caffeine, but did not affect the size of the thapsigargin-sensitive Ca2+ stores. The other substitutions had no effect on either the size of the intracellular Ca2+ stores, or on the amount of Ca2+ released after ryanodine receptor activation; however, both the P3527S and V4849I substitutions had a small but significant effect on the resting Ca2+ concentration.

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Functional Characterization of C-terminal Ryanodine Receptor 1 Variants Associated with Central Core Disease or Malignant Hyperthermia

Journal of Neuromuscular Diseases ◽

10.3233/jnd-170210 ◽

2017 ◽

Vol 4 (2) ◽

pp. 147-158 ◽

Cited By ~ 3

Author(s):

Remai Parker ◽

Anja H. Schiemann ◽

Elaine Langton ◽

Terasa Bulger ◽

Neil Pollock ◽

...

Keyword(s):

Malignant Hyperthermia ◽

Ryanodine Receptor ◽

Functional Characterization ◽

Central Core ◽

Central Core Disease ◽

Ryanodine Receptor 1

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Results of Contracture Tests with Halothane, Caffeine, and Ryanodine Depend on Different Malignant Hyperthermia-associated Ryanodine Receptor Gene Mutations

Anesthesiology ◽

10.1097/00000542-200208000-00010 ◽

2002 ◽

Vol 97 (2) ◽

pp. 345-350 ◽

Cited By ~ 16

Author(s):

Marko Fiege ◽

Frank Wappler ◽

Ralf Weisshorn ◽

Mark Ulrich Gerbershagen ◽

Markus Steinfath ◽

...

Keyword(s):

Malignant Hyperthermia ◽

Ryanodine Receptor ◽

Time Course ◽

Clinical Symptoms ◽

Receptor Gene ◽

Gene Mutations ◽

Onset Time ◽

Release Channel ◽

Ryr1 Gene

Background More than 20 mutations in the gene encoding for the ryanodine receptor (RYR1), a Ca2+ release channel of the skeletal muscle sarcoplasmic reticulum, have been found to be associated with malignant hyperthermia (MH). This study was designed to investigate the effects of different mutations in the RYR1 gene on contracture development in in vitro contracture tests (IVCT) with halothane, caffeine, and ryanodine. Methods Ninety-three MH-susceptible (MHS) patients, diagnosed by the standard IVCT with halothane and caffeine, were included in this prospective study. Surplus muscle specimens were used for an IVCT with 1 microm ryanodine. The contracture course during the ryanodine IVCT was described by the attainment of different time points: onset time of contracture and times when contracture reached 2 mN or 10 mN. In addition, all patients were screened for mutations of the RYR1 gene. Results In 36 patients, four different mutations of the RYR1 gene (C487-T, G1021-A, C1840-T, G7300-A) were found. The IVCT threshold concentrations of halothane and caffeine were lower in patients with the C487-T mutation compared with patients without a detected mutation in the RYR1 gene. In the IVCT with ryanodine, contracture levels of 2 mN and 10 mN were reached earlier in muscle specimens from patients with C487-T, C1840-T, and G7300-A mutations compared with specimens from patients with the G1021-A mutation and patients without detected mutation in the RYR1 gene. Conclusions The differences between the groups in the halothane and caffeine IVCT threshold concentrations and in the time course of contracture development in the ryanodine IVCT underline the hypothesis that certain mutations in the RYR1 gene could make the ryanodine receptor more sensitive to specific ligands. This may be an explanation for varying clinical symptoms of MH crisis in humans.

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