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Table 3 Cellular RYR1 model systems: Immortalized B-lymphocytes

From: Preclinical model systems of ryanodine receptor 1-related myopathies and malignant hyperthermia: a comprehensive scoping review of works published 1990–2019

Author/Year RYR1 variant(s) Title Conclusions
Zullo A, et al. [110] 2019 R1335C, S2345R, S3098I, F4924_V4925ins RYR1 sequence variants in myopathies: Expression and functional studies in two families Ca2+ release in response to the RyR1 agonist 4-chloro-m-cresol and to thapsigargin showed that S2345R causes depletion of S/ER Ca2+ stores and that the compound heterozygosity for variant S3098I and the 30-nucleotide insertion increases RyR1-dependent Ca2+ release without affecting ER Ca2+ stores
Johannsen S, et al. [111] 2016 R4737W Functional characterization of the RYR1 mutation pArg4737Trp associated with susceptibility to malignant hyperthermia Intracellular resting calcium was slightly but significantly elevated in mutation positive cells. Calcium release following stimulation with 4-chloro-m-cresol was significantly increased in B lymphocytes carrying the R4737W mutation compared to mutation negative controls
Schiemann AH, et al. [112] 2014 R2355W, V2354M Functional characterization of 2 known ryanodine receptor mutations causing malignant hyperthermia We propose that R2355W is confirmed as being an MH-causative mutation and suggest that V2354M is a RYR1 mutation likely to cause MH
Lyfenko AD, et al. [66] 2007 R4214_F4216del Two central core disease (CCD) deletions in the C-terminal region of RYR1 alter muscle excitation-contraction (EC) coupling by distinct mechanisms ΔRQF RYR1 deletion did not significantly affect the sensitivity of lymphoblastoid cells to activation by 4-CmC. However, Ca21 release activated by a maximal concentration of 4-CmC (1 mM) was significantly reduced in ΔRQF-carrying cells
Schiemann AH, et al. 2013 R1583C, V2102L Sequence capture and massively parallel sequencing to detect mutations associated with malignant hyperthermia The amount of Ca2+ released after stimulation with 4-chloro-m-cresol from B lymphocytes of the MH-susceptible patients in the family was significantly greater compared with that of Ca2+ released from cells of an MH-negative family member
Attali R, et al. [113] 2013 Y3016C Variable myopathic presentation in a single family with novel skeletal RYR1 mutation Functional analysis on EBV immortalized cell lines showed no effect of the mutation on RyR1 pharmacological activation or the content of intracellular Ca2+ stores
Vukcevic M, et al. [114] 2010 R1679H, K1393R, E1058K, H382N, R2508G Functional properties of RYR1 mutations identified in Swedish patients with malignant hyperthermia and central core disease All B lymphoblastoid cell lines carrying RYR1 candidate mutations showed significantly increased resting cytoplasmic Ca2+ levels as well as a shift to lower concentrations of 4-CmC inducing calcium release compared with controls
Grievink H, et al. [115] 2010 H4833Y Allele-specific differences in ryanodine receptor 1 mRNA expression levels may contribute to phenotypic variability in malignant hyperthermia Allele-specific differences in RYR1 mRNA expression levels in heterozygous MHS samples, and can at least in part contribute to the observed variable penetrance and variations in MH clinical phenotypes
Zozato F, et al. [72] 2003 F4863_D4869delinsT Clinical and functional effects of a deletion in a COOH-terminal lumenal loop of the skeletal muscle ryanodine receptor Cells have depleted thapsigargin-sensitive intracellular Ca2+ stores, exhibit release of Ca2+ from intracellular stores in the absence of the addition of a pharmacological activator of the RYR1; and the unelicited Ca2+ transient from the thapsigargin-sensitive stores could be blocked by dantrolene, a specific inhibitor of the skeletal muscle RYR
Zullo A, et al. [116] 2009 R530H, R2163P, N2342S, E2371G, R2454H, C4664R Functional characterization of ryanodine receptor (RYR1) sequence variants using a metabolic assay in immortalized B-lymphocytes Increased acidification rate of lymphoblastoid cells in response to 4-CmC is mainly due to RYR1 activation. Cells expressing RYR1 variants in the N-terminal and in the central region of the protein (R530H, R2163P, N2342S, E2371G and R2454H) displayed higher activity compared with controls. Cell lines harboring RYR1(C4664R) were significantly less activated by 4-CmC
Levano S, et al. [117] 2009 D554Y, R2336H, E2404K, D2730G Increasing the number of diagnostic mutations in malignant hyperthermia All RYR1 mutations significantly increased resting calcium concentration and significantly affect either 4-CmC or caffeine dose-response curve to pharmacological activation Only one mutation (D2730G) showed a significant reduction in EC50 of both caffeine and 4-CmC
Anderson AA, et al. [118] 2008 H4833Y Identification and biochemical characterization of a novel ryanodine receptor gene mutation associated with malignant hyperthermia B lymphocytes from patients with this mutation were approximately twofold more sensitive than MH-negative cells to activation with 4-CmC. The amount of Ca2+ released from B lymphocytes of MH-susceptible patients was significantly greater than that released from cells of family members without this mutation
Ducreux D, et al. [119] 2006 P3527S, V4849I, R999H 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 P3527S in the homozygous state affected the amount of Ca2+ released after pharmacological activation with 4-CmC and caffeine but did not affect the size of the thapsigargin-sensitive Ca2+ stores. The other substitutions had no effect on the size of the intracellular Ca2+ stores, or the amount of Ca2+ released after ryanodine receptor activation P3527S and V4849I substitutions had a small but significant effect on the resting Ca2+ concentration
Tilgen N, et al. [120] 2001 R4861H, I4898T, G4899A Identification of four novel mutations in the C-terminal membrane spanning domain of the ryanodine receptor 1: Association with central core disease and alteration of calcium homeostasis Cell showed release of Ca2+ from intracellular stores in the absence of any pharmacological activators of RYR, significantly smaller thapsigargin-sensitive intracellular calcium stores, compared to lymphoblasts from control individuals, and a normal sensitivity of the calcium release to the RYR inhibitor dantrolene
Girard T, et al. [121] 2001 V2168M B-lymphocytes from malignant hyperthermia-susceptible patients have an increased sensitivity to skeletal muscle ryanodine receptor activators EBV immortalized cells harboring the V2168M RYR1 gene mutation were more sensitive to the RYR activator 4-CmC and their peripheral blood leukocytes produce more interleukin-1beta after treatment with the RYR activators caffeine and 4-CmC, compared with cells from healthy controls
Hoppe K, et al. [122] 2016 R530H, C4664R, R2163P Hypermetabolism in B–lymphocytes from malignant hyperthermia susceptible individuals Native B–lymphocytes from MHS individuals are more sensitive to 4–CmC than those from MHN, reflecting a greater Ca2+ turnover. The acidification response, however, was less pronounced than in muscle cells, presumably reflecting the lower expression of RyR1 in B–lymphocytes.