A novel RyR1-selective inhibitor prevents and rescues sudden death in mouse models of malignant hyperthermia and heat stroke

Yamazawa, Toshiko; Kobayashi, Takuya; Kurebayashi, Nagomi; Konishi, Masato; Noguchi, S.; Inoue, Tohru; Inoue, Yukiko; Nishino, Ichizo; Mori, Shigeo; Iinuma, Hiroto; Kagechika, Hiroyuki; Uryash, Arkady; Adams, José A.; López, José R.; Liu, Xiaochen; Diggle, Christine P.; Allen, Paul D.; Kakizawa, Sho; Ikeda, Kenro; Lin, Bangzhong; Ikemi, Yui; Nunomura, Kazuto; Nakagawa, Shinsaku; Sakurai, Takashi; Murayama, Takashi

doi:10.1038/s41467-021-24644-1

Cited by 34 publications

(33 citation statements)

References 53 publications

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“…6). This is consistent with the suppression of twitch and tetanic tensions of isolated muscles and of reduced in vivo muscle weakness by dantrolene 46,47 and Cpd1 24 . Interestingly, procaine did not suppress DICR (Fig.…”

Section: Discussionsupporting

confidence: 87%

Reconstituted depolarization-induced Ca²⁺release platform for skeletal muscle disease mutation validation and drug discovery

Murayama

Kurebayashi

Numaga‐Tomita

et al. 2022

Preprint

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In skeletal muscle excitation-contraction (E-C) coupling, depolarization of the plasma membrane triggers Ca2+ release from the sarcoplasmic reticulum (SR), referred to as depolarization-induced Ca2+ release (DICR). DICR occurs via the type 1 ryanodine receptor (RyR1), which physically interacts with the dihydropyridine receptor Cav1.1 subunit in specific machinery formed with additional components. Exome sequencing has accelerated the discovery of many novel mutations in DICR machinery genes in various skeletal muscle diseases. However, functional validation is time-consuming because it must be performed in a skeletal muscle environment. Here, we describe a high-throughput platform for DICR that is reconstituted in non-muscle HEK293 cells. We demonstrate that Cav1.1 mutations implicated in malignant hyperthermia and myopathy exhibit divergent effects on DICR. We also tested several RyR1 inhibitors and DICR modulators. This high-throughput DICR platform will accelerate validation of mutations and drug discovery for skeletal muscle diseases.

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Section: Discussionsupporting

confidence: 87%

Reconstituted depolarization-induced Ca²⁺release platform for skeletal muscle disease mutation validation and drug discovery

Murayama

Kurebayashi

Numaga‐Tomita

et al. 2022

Preprint

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“…Variants within the RYR1 gene have a causative role in the development of malignant hyperthermia (MH), a rare, but serious condition characterized by sustained muscle contraction, hyperthermia, and rhabdomyolysis in response to halogenated anesthetics [ 15 ]. Dantrolene is a RYR antagonist that stabilizes receptor resting state by increasing Mg 2+ affinity and is currently the only FDA approved drug for MH, although a RYR1-selective inhibitor, 6,7-(methylenedioxy)-1-octyl-4-quinolone-3-carboxylic acid, was recently shown to be effective in murine MH models [ 20 , 22 ]. Henderson et al, 2021 recently described a high throughput screen using the exodosis reporter Gaussia luciferase-secreted ER calcium modulated protein (GLuc-SERCaMP) and identified several FDA approved drugs effective in attenuating exodosis including dantrolene [ 10 ].…”

Section: Resultsmentioning

confidence: 99%

Identification of ER/SR resident proteins as biomarkers for ER/SR calcium depletion in skeletal muscle cells

Greer

Meilleur

Harvey

et al. 2022

Orphanet J Rare Dis

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Background Aberrations to endoplasmic/sarcoplasmic reticulum (ER/SR) calcium concentration can result in the departure of endogenous proteins in a phenomenon termed exodosis. Redistribution of the ER/SR proteome can have deleterious effects to cell function and cell viability, often contributing to disease pathogenesis. Many proteins prone to exodosis reside in the ER/SR via an ER retention/retrieval sequence (ERS) and are involved in protein folding, protein modification, and protein trafficking. While the consequences of their extracellular presence have yet to be fully delineated, the proteins that have undergone exodosis may be useful for biomarker development. Skeletal muscle cells rely upon tightly coordinated ER/SR calcium release for muscle contractions, and perturbations to calcium homeostasis can result in myopathies. Ryanodine receptor type-1 (RYR1) is a calcium release channel located in the SR. Mutations to the RYR1 gene can compromise calcium homeostasis leading to a vast range of clinical phenotypes encompassing hypotonia, myalgia, respiratory insufficiency, ophthalmoplegia, fatigue and malignant hyperthermia (MH). There are currently no FDA approved treatments for RYR1-related myopathies (RYR1-RM). Results Here we examine the exodosis profile of skeletal muscle cells following ER/SR calcium depletion. Proteomic analysis identified 4,465 extracellular proteins following ER/SR calcium depletion with 1,280 proteins significantly different than vehicle. A total of 54 ERS proteins were identified and 33 ERS proteins significantly increased following ER/SR calcium depletion. Specifically, ERS protein, mesencephalic astrocyte-derived neurotrophic factor (MANF), was elevated following calcium depletion, making it a potential biomarker candidate for human samples. Despite no significant elevation of MANF in plasma levels among healthy volunteers and RYR1-RM individuals, MANF plasma levels positively correlated with age in RYR1-RM individuals, presenting a potential biomarker of disease progression. Selenoprotein N (SEPN1) was also detected only in extracellular samples following ER/SR calcium depletion. This protein is integral to calcium handling and SEPN1 variants have a causal role in SEPN1-related myopathies (SEPN1-RM). Extracellular presence of ER/SR membrane proteins may provide new insight into proteomic alterations extending beyond ERS proteins. Pre-treatment of skeletal muscle cells with bromocriptine, an FDA approved drug recently found to have anti-exodosis effects, curbed exodosis of ER/SR resident proteins. Conclusion Changes to the extracellular content caused by intracellular calcium dysregulation presents an opportunity for biomarker development and drug discovery.

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“…Dnajb4 KI and KO mouse lines were generated via zygote electroporation utilizing the CRISPR/Cas9 system described previously 50, 51 . We designed the CRISPR RNA (crRNA) to target the coding sequences near c.270C > A (p.F90L) (Supplementary Fig.…”

Section: Methodsmentioning

confidence: 99%

Distinctive chaperonopathy in skeletal muscle associated with the dominant variant inDNAJB4

Inoue

Noguchi

Inoue

et al. 2022

Preprint

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DnaJ homolog, subfamily B, member 4, a member of the heat shock protein 40 chaperones encoded by DNAJB4, is highly expressed in myofibers. We identified a heterozygous c.270 T>A (p.F90L) variant in DNAJB4 in a family with a dominantly inherited distal myopathy, in which affected members have specific features on muscle pathology represented by the presence of cytoplasmic inclusions and the accumulation of desmin, p62, HSP70 and DNAJB4 predominantly in type 1 fibers. Both Dnajb4-F90L knock-in and knockout mice developed muscle weakness and recapitulated the patient muscle pathology in the soleus muscle, where DNAJB4 has the highest expression. These data indicate that the identified variant is causative resulting in defective chaperone function and selective muscle degeneration in specific muscle fibers. This study demonstrates the importance of DNAJB4 in skeletal muscle proteostasis by identifying the associated chaperonopathy.

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A novel RyR1-selective inhibitor prevents and rescues sudden death in mouse models of malignant hyperthermia and heat stroke

Cited by 34 publications

References 53 publications

Reconstituted depolarization-induced Ca²⁺release platform for skeletal muscle disease mutation validation and drug discovery

Reconstituted depolarization-induced Ca²⁺release platform for skeletal muscle disease mutation validation and drug discovery

Identification of ER/SR resident proteins as biomarkers for ER/SR calcium depletion in skeletal muscle cells

Distinctive chaperonopathy in skeletal muscle associated with the dominant variant inDNAJB4

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A novel RyR1-selective inhibitor prevents and rescues sudden death in mouse models of malignant hyperthermia and heat stroke

Cited by 34 publications

References 53 publications

Reconstituted depolarization-induced Ca2+release platform for skeletal muscle disease mutation validation and drug discovery

Reconstituted depolarization-induced Ca2+release platform for skeletal muscle disease mutation validation and drug discovery

Identification of ER/SR resident proteins as biomarkers for ER/SR calcium depletion in skeletal muscle cells

Distinctive chaperonopathy in skeletal muscle associated with the dominant variant inDNAJB4

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Reconstituted depolarization-induced Ca²⁺release platform for skeletal muscle disease mutation validation and drug discovery

Reconstituted depolarization-induced Ca²⁺release platform for skeletal muscle disease mutation validation and drug discovery