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Journal of Anesthesia

, Volume 32, Issue 2, pp 174–181 | Cite as

Genetic and functional analysis of the RYR1 mutation p.Thr84Met revealed a susceptibility to malignant hyperthermia

  • Takashi Kondo
  • Toshimichi Yasuda
  • Keiko Mukaida
  • Sachiko Otsuki
  • Rieko Kanzaki
  • Hirotsugu Miyoshi
  • Hiroshi Hamada
  • Ichizo Nishino
  • Masashi Kawamoto
Original Article

Abstract

Purpose

The aim of this study was to analyze the genetic and functional role of a novel RYR1 variant c.251 C > T (p.Thr84Met) identified in a patient with muscle weakness demonstrating MH susceptibility.

Methods

DNA testing of family members was conducted for assessment of pathogenicity of the genetic variant. For functional analysis, Ca2+ measurement using patient-derived myotubes and p.Thr84Met RYR1-transfected human embryonic kidney (HEK)-293 cells was performed to evaluate reactivity to RYR1 activators. The half-maximal effective concentration (EC50) values of two RYR1 activators, caffeine and 4-chloro-m-cresol (4CmC), were calculated from the acquired dose–response curves. The EC50 was compared between two groups: for myotubes, the control group and the patient, and for HEK-293 cells, WT and p.Thr84Met.

Results

Dose–response curves for caffeine and 4CmC were shifted to the left in both myotubes and HEK-293 cells compared to controls. The 50% effective concentration values for caffeine and 4CmC were significantly lower in both myotubes and HEK-293 cells compared to controls (P < 0.001 for all comparisons).

Conclusions

Our results of functional testing indicated RYR1 hypersensitivity to caffeine and 4CmC. We conclude that the genetic variant was associated with MH susceptibility.

Keywords

Malignant hyperthermia Ryanodine receptor Mutation Calcium release 

Notes

Acknowledgements

We thank Professor David H. MacLennan for the generous gift of RYR1/pcDNA.

Compliance with ethical standards

Conflict of interest

This study was supported in part by a Grant-in-Aid (Grant number 16K20098) for Scientific Research from the Japan Society for the Promotion of Science, Japan.

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Copyright information

© Japanese Society of Anesthesiologists 2018

Authors and Affiliations

  1. 1.Department of Anesthesiology and Critical CareHiroshima University HospitalHiroshimaJapan
  2. 2.Department of Anesthesiology and Critical Care, Graduate School of Biomedical SciencesHiroshima UniversityHiroshimaJapan
  3. 3.Department of AnesthesiologyHiroshima Prefectural Rehabilitation CenterHiroshimaJapan
  4. 4.Department of Neuromuscular ResearchNational Institute of Neuroscience, National Center of Neurology and PsychiatryKodairaJapan

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