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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