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Table 1 Cellular RYR1 model systems: Human embryonic kidney (HEK-293) cells

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
HEK-293
 Chirasani VR, et al. [34] 2019 Q3970K, Q3970E A central core disease mutation in the Ca2+ binding site of skeletal muscle ryanodine receptor impairs single channel regulation RyR1-Q3970K is likely a CCD-associated loss-of-function channel that conducts Ca2+
 Xu L, et al. [35] 2018 G4934D, G4934K, G4941D, G4941K, G4941M, D4938N, D4945N G4941K substitution in the pore-lining S6 helix of the skeletal muscle ryanodine receptor increases RyR1 sensitivity to cytosolic and luminal Ca2+ Luminal Ca2+ accesses Ca2+ activation sites as they pass through the pore rather than traveling to openings that lie outside the pore
 Xu L, et al. [36] 2018 E3893Q, E3893V, E3967Q, E3967V, T5001A Ca2+ − mediated activation of the skeletal-muscle ryanodine receptor ion channel Removal of negative charges in a RyR1 Ca2+ binding site impairs activation of RyR1 by physiological concentrations of Ca2+, and suggests loss of binding to or reduced Ca2+ affinity of the site
 Xu L, et al. [37] 2008 G4898E, G4898R, ΔV4926, ΔI4927, R110W, L486V Single channel properties of heterotetrameric mutant RyR1 ion channels linked to core myopathies Homozygous RyR1 mutations associated with core myopathies abolish or greatly reduce sarcoplasmic reticulum Ca2+ release during excitation-contraction coupling
 Schiemann AH, et al. [38] 2018 D2431Y A genetic mystery in malignant hyperthermia ‘solved’? The D2431Y variant is pathogenic for MH and should be added to the European Malignant Hyperthermia Group (EMHG) list of diagnostic mutations
 Murayama T, et al. [39] 2018 G342R, R2435H, L4824P Efficient High-Throughput Screening by Endoplasmic Reticulum Ca2+ Measurement to Identify Inhibitors of Ryanodine Receptor Ca2+ − Release Channels In the current high throughput screening of 1535 compounds, we identified four RyR1 inhibitors
 Kondo T, et al. [40] 2018 T84M Genetic and functional analysis of the RYR1 mutation pThr84Met revealed a susceptibility to malignant hyperthermia Functional analysis of T84M demonstrated higher responsivity to caffeine and 4CmC
 Parker R, et al. [41] 2017 M4640I, V4849I, F4857S, D4918N Functional Characterization of C-terminal Ryanodine Receptor 1 Variants Associated with Central Core Disease or Malignant Hyperthermia The V4849I variant should be considered a risk factor for malignant hyperthermia, while the F4857S and D4918N variants should be classified as pathogenic for CCD
 Merritt A, et al. [42] 2017 R2336H, R2355W, E3104K, G3990V, V4849I, D3986E Assessing the pathogenicity of RYR1 variants in malignant hyperthermia Functional analyses in HEK293 cells provided evidence to support the use of R2336H, R2355W, E3104K, pG3990V and V4849I for diagnostic purposes but not D3986E
 Chen W, et al. [43] 2017 R164C, Y523S, R2136H, R2435H, Y4796C Reduced threshold for store overload-induced Ca2+ release is a common defect of RyR1 mutations associated with malignant hyperthermia and central core disease All mutations reduced the threshold for SOICR
 Stephens J et al. [44] 2016 ΔE2348, T214M Functional analysis of RYR1 variants linked to malignant hyperthermia ΔE2348 could be added to the list of diagnostic mutations for susceptibility to malignant hyperthermia T214M, does not appear to significantly alter sensitivity to agonist in the same system
 Murayama T, et al. [45] 2016 R2163C, R2163H, V2168M, T2206M, A2350T, G2375A, G2434R, R2435H, R2454C, R2454H, R2458C, R2458H, R2508C, R2508H Genotype–Phenotype Correlations of Malignant Hyperthermia and Central Core Disease Mutations in the Central Region of the RYR1 Channel In live-cell Ca2+ imaging, the mutant channels exhibited an enhanced sensitivity to caffeine, a reduced endoplasmic reticulum Ca2+ content, and an increased resting cytoplasmic Ca2+ level
 Gomez AC, et al. [46] 2016 F4732D, G4733E, R4736W, R4736Q, T4825I, H4832Y, T4082M, S4113L, N4120Y Malignant hyperthermia-associated mutations in the S2-S3 cytoplasmic loop of type 1 ryanodine receptor calcium channel impair calcium-dependent inactivation Nine RyR1 mutants associated with skeletal muscle diseases were differently regulated by Ca2+ and Mg2+
 Murayama T, et al. [47] 2015 C36R, R164C, R164L, G249R, G342R, R402C, R402H, Y523C, Y523S, R615C, R615L Divergent Activity Profiles of Type 1 Ryanodine Receptor Channels Carrying Malignant Hyperthermia and Central Core Disease Mutations in the Amino-Terminal Region The mutations increased the gain and the sensitivity to activating Ca2+ in a site-specific manner. Gain was consistently higher in both MH and MH/CCD mutations
 Miyoshi H, et al. [48] 2015 R2508H, R2508G, R2508S, R2508K Several Ryanodine Receptor Type 1 Gene Mutations of pArg2508 Are Potential Sources of Malignant Hyperthermia Cells transfected with each of the 4 mutants, R2508H, R2508G, R2508S, or R2508K, were more sensitive to caffeine and 4CmC than wild-type cells
 Mei Y, et al. [49] 2015 G4934A, G4934V, G4941V, G4941A, G4941I Channel Gating Dependence on Pore Lining Helix Glycine Residues in Skeletal Muscle Ryanodine Receptor Both glycines are important for RyR1 channel function by providing flexibility and minimizing amino acid clashes
 Shirvanyants D, et al. [50] 2014 M4887G, M4887A, M4887V, V4891A, I4897Y Pore dynamics and conductance of RyR1 transmembrane domain Loss of these interactions in the case of polar substitution I4897T results in destabilization of the selectivity filter, a possible cause of the CCD-specific reduced Ca2+ conductance
 Roesl C, et al. [51] 2014 R2452W Functional characterisation of the R2452W ryanodine receptor variant associated with malignant hyperthermia susceptibility R2452W results in a hypersensitive ryanodine receptor 1 and is likely to be causative of MH
 Miyoshi H, et al. [52] 2014 R2508C, R2508H, R2508K, R2508S Two different variants of p.2508 in Japanese malignant hyperthermia patients causing hypersensitivity of ryanodine receptor 1 All alterations in the p.2508 portion of RyR1 play important roles in the pathogenesis of MH
 Sato K, et al. [53] 2013 R44C, R163C, R401C, R533C, R533H, H4833Y Skeletal muscle ryanodine receptor mutations associated with malignant hyperthermia showed enhanced intensity and sensitivity to triggering drugs when expressed in human embryonic kidney cells These six mutations cause functional abnormality of the calcium channel, leading to higher sensitivity to a specific agonist
 Kraeva N, et al. [54] 2013 M4640R, L4647P, F4808L, D4918N, F4941C Novel excitation-contraction uncoupled RYR1 mutations in patients with central core disease Homotetrameric RyR1 mutants harbouring L4646P, F4807P, D4917N and R4892Q mutations abolished caffeine-induced Ca2+ release
 Merritt A, et al. [55] 2012 D1056H Functional analysis of the pD1056H RYR1 variant associated with malignant hyperthermia and exertional heat stroke Cells expressing D1056H exhibited a trend for greater calcium release and increased sensitivity than wild-type at low doses of caffeine
 Murayama T, et al. [56] 2011 T4825A, T4825I, I4826A, L4827A, S4828A, S4829A Role of amino-terminal half of the S4-S5 linker in type 1 ryanodine receptor (RyR1) channel gating Four mutants had reduced CICR activity without changing Ca2+ sensitivity, whereas the L4827A mutant formed a constitutive active channel T4825I, a disease-associated mutation for malignant hyperthermia, exhibited enhanced CICR activity
 Haraki T, et al. [57] 2011 A4894T, A4894P, A4894S, A4894G Mutated p.4894 RyR1 function related to malignant hyperthermia and congenital neuromuscular disease with uniform type 1 fiber (CNMDU1) The hypersensitive A4894T-RyR1 is associated with MH and the poorly functional A4894P-RyR1 with CNMDU1
 Zhou H, et al. [58] 2010 R2939K Multi-minicore disease and atypical periodic paralysis associated with novel mutations in the skeletal muscle ryanodine receptor (RYR1) gene The R2435K mutation did not affect two characteristic functional properties of RyR1, as both Ca2+ dependence and activation by caffeine were not altered
 Sato K, et al. [59] 2010 R163C, G248R, T4826I, H4833Y, I4898T, G4899R Functional studies of RYR1 mutations in the skeletal muscle ryanodine receptor using human RYR1 complementary DNA MH mutations showed a higher response, whereas CCD mutants (I4898T and G4899R) did not respond to 4-Cm C
 Merritt A, et al. [60] 2010 G3990V Functional analysis of the pGly3990Val RYR1 variant using a human cDNA clone in HEK293 cells A statistically significant increase in Ca2+ release was observed in G3990V mutants at each caffeine concentration that elicited a response
 Migita T, et al. [61] 2009 R2508C Functional analysis of ryanodine receptor type 1 pR2508C mutation in exon 47 The transfected RYR1 mutant was more sensitive to caffeine and 4CmC than wildtype RYR1
 Migita T, et al. [62] 2009 R2508C, A4894T Do Ca2+ channel blockers improve malignant hyperthermia crisis? The dantrolene-induced decline effect of Ca2+ of skeletal muscle was not disappeared in the presence of Ca2+ blockers. In MH crisis, we do not recommend to administer Ca2+ blockers because of its potent effect to increase Ca2+
 Ghassemi F, et al. [63] 2009 R2435L A recessive ryanodine receptor 1 mutation in a CCD patient increases channel activity R2435L does not affect resting Ca2+, or sensitivity of RyR1 to pharmacological activators Instead it reduces the release of Ca2+ from intracellular stores induced by pharmacological activators as well as by KCl via the voltage sensing dihydropyridine receptor
 Jiang D, et al. [64] 2008 R615C Reduced threshold for luminal Ca2+ activation of RyR1 underlies a causal mechanism of porcine malignant hyperthermia R615C confers MH susceptibility by reducing the threshold for luminal Ca2+ activation and SOICR
 Rossi D, et al. [65] 2007 R4836fsX4838 A truncation in the RYR1 gene associated with central core lesions in skeletal muscle fibres Subtle changes in Ca2+ release of human heteromeric RyR1/RyR1R4837fsX4839 channels, probably due to the reduced stability/assembly of these channels, may predispose individuals to MHS
 Lyfenko AD, et al. [66] 2007 R4214_F4216del, V4927_I4928del Two central core disease (CCD) deletions in the C-terminal region of RYR1 alter muscle excitation-contraction (EC) coupling by distinct mechanisms Single channel data indicate that the ΔRQF mutation increases Ca2+ responsiveness without altering K+ conductance and ion selectivity for Ca2+ compared to K+. In contrast, the ΔVI deletion abolished Ca2+ responsiveness, Ca2+ permeation, and significantly reduced K+ conductance demonstrating that the ΔVI mutation introduced major alterations to the channel pore
 Zhou H, et al. [67] 2006 S71Y, R110W, L486V, A1578T, S2060C, N2283H Characterization of recessive RYR1 mutations in core myopathies Recombinant channels with N2283H substitution showed an increased activity, whereas recombinant channels with S71Y + N2283H substitution lost activity upon isolation
 Xu L, et al. [68] 2006 D4938N, D4945N, D4953N, E4942Q, E4948Q, E4952Q, E4955Q Two rings of negative charges in the cytosolic vestibule of type-1 ryanodine receptor modulate ion fluxes D4938N and D4945N exhibited an attenuated block by neomycin to a greater extent from the cytosolic than lumenal side. By comparison, charge neutralization of lumenal loop residues (D4899Q, E4900N) eliminated the block from the lumenal but not the cytosolic side
 Wang Y, et al. [69] 2005 D4899Q, E4900N Probing the role of negatively charged amino acid residues in ion permeation of skeletal muscle ryanodine receptor the negatively charged carboxyl oxygens of D4899 and E4900 side chains are major determinants of RyR ion conductance and selectivity
 Brini M, et al. 2005 [70] R615C, Y523S, I4898T Ca2+ signaling in HEK-293 and skeletal muscle cells expressing recombinant ryanodine receptors harboring malignant hyperthermia and central core disease mutations I4898T RyR1 channels produced cytosolic Ca2+ values which were similar to those observed for WT RyR1 channels R615C augmented the amplitude of the cytosolic and mitochondrial Ca2+ transients following cell stimulation By contrast, the mitochondrial Ca2+ transients were reduced in cells expressing Y523S
 Du GG, et al. [71] 2004 R4892W, I4897T, G4898E Central core disease mutations R4892W, I4897T and G4898E in the ryanodine receptor isoform 1 reduce the Ca2+ sensitivity and amplitude of Ca2+-dependent Ca2+ release Ca2+ sensitivity is one of the serious defects in these three excitation-contraction uncoupling CCD mutations
 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 Channels carrying the deletion were less stable than the wild-type channels and disappeared rapidly when recorded at membrane potentials greater than ±20 mV
 Stange M, et al. [73] 2003 S2843D, S2843A Characterization of recombinant skeletal muscle (Ser-2843) and cardiac muscle (Ser-2809) ryanodine receptor phosphorylation mutants Results did not support the view that phosphorylation of a single site (RyR1-Ser-2843 and RyR2-Ser-2809) substantially changes RyR1 and RyR2 channel function
 Loke JC, et al. [74] 2003 R328W Detection of a novel ryanodine receptor subtype 1 mutation (R328W) in a malignant hyperthermia family by sequencing of a leukocyte transcript The mutant channel has increased sensitivity to both caffeine and halothane
 Yamaguchi N, et al. [75] 2001 V3619A, W3620A, L3624D, Δ4274–4535 Identification of apocalmodulin and Ca2+ − calmodulin regulatory domain in skeletal muscle Ca2+ release channel, ryanodine receptor Two single amino acid substitutions distinctly change the regulation of the skeletal muscle Ca2+ release channel by CaM; one of which (L3624D) results in a loss of activation by apoCaM and an inhibition by CaCaM, whereas the other (W3620A) specifically abolishes CaCaM inhibition RyR1Δ4274–4535, showed an ∼10-fold increased sensitivity to activating Ca2+
 Sun J, et al. [76] 2001 C3635A Cysteine-3635 is responsible for skeletal muscle ryanodine receptor modulation by NO C3635A resulted in the loss of CaM-dependent NO modulation of channel activity and reduced S-nitrosylation by NO to background levels but did not affect NO-independent channel modulation by CaM or the redox sensitivity of the channel to O(2) and glutathione
 Gaburjakova M, et al. [77] 2001 V2461H, V2461E, V2461G, V2461I FKBP12 binding modulates ryanodine receptor channel gating Val2461 is a critical residue required for FKBP12 binding to RyR1 FKBP12 has a functional role in the RyR1 channel complex
 Du GG, et al. [78] 2001 G2370A, G2372A, G2373A, G2375A, Y3937A, S3938A, G3939A, K3940A Mutations to Gly2370, Gly2373 or Gly2375 in malignant hyperthermia domain 2 decrease caffeine and cresol sensitivity of the rabbit skeletal-muscle Ca2+ − release channel (ryanodine receptor isoform 1) Amino acids 2370–2375 lie within a sequence (amino acids 2163–2458) in which 8 RyR1 mutations associated with MH have been shown to be hypersensitive to caffeine and 4-chloro-m-cresol activation By contrast, G2370A, G2373A and G2375A are hyposensitive to caffeine and 4-chloro-m-cresol Amino acids 2163–2458 form a regulatory domain (MH regulatory domain 2) that regulates caffeine and 4-chloro-m-cresol sensitivity of RyR1
 Monnier N, et al. [79] 2000 Y4796C An autosomal dominant congenital myopathy with cores and rods is associated with a neomutation in the RYR1 gene encoding the skeletal muscle ryanodine receptor Expression of the mutant RYR1 cDNA produced channels with increased caffeine sensitivity and a significantly reduced maximal level of Ca2+ release Single-cell Ca2+ analysis showed that the resting cytoplasmic level was increased by 60% in cells expressing the mutant channel
 Gao L, et al. [80] 2000 I4897A, I4897L, I4897V, D4917A, D4899A, D4899R, R4913E, G4894A, D4899N Evidence for a role of the lumenal M3-M4 loop in skeletal muscle Ca2+ release channel (ryanodine receptor) activity and conductance Amino acid residues in the lumenal loop region between the two most C-terminal membrane segments constitute a part of the ion-conducting pore of RyR1
 Tong J, et al. [81] 1999 C36R, G249R, G342R, R553W, R615R, R615C, R2163C, G2435R, R2458C, R2458H, R164C, I404M, Y523S, R2163H, R2436H Measurement of resting cytosolic Ca2+ concentrations and Ca2+ store size in HEK-293 cells transfected with malignant hyperthermia or central core disease mutant Ca2+ release channels MH/CCD mutants were more sensitive to caffeine than WT RyR1, indicating that caffeine hypersensitivity observed with a variety of MH/CCD mutant RyR1 proteins is not dependent on extracellular Ca2+ concentration
 Lynch PJ, et al. [82] 1999 I4898T A mutation in the transmembrane/luminal domain of the ryanodine receptor is associated with abnormal Ca2+ release channel function and severe central core disease Single-cell analysis of co-transfected cells showed a significantly increased resting cytoplasmic Ca2+ level and a significantly reduced luminal Ca2+ level These data are indicative of a leaky channel, possibly caused by a reduction in the Ca2+ concentration required for channel activation
 Tong J, et al. [83] 1997 R164C, G249R, G342R, I404M, Y523S, R615C, G2435R, R2436H, C36R, R553W, R615L, R2163C, R2163H, R2458C, R2458H Caffeine and halothane sensitivity of intracellular Ca2+ release is altered by 15 calcium release channel (ryanodine receptor) mutations associated with malignant hyperthermia and/or central core disease Abnormal sensitivity in the Ca2+ photometry assay provides supporting evidence for a causal role in MH for each of 15 single amino acid mutations in the ryanodine receptor
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Orphanet Journal of Rare Diseases

ISSN: 1750-1172