NAD+ dyshomeostasis in RYR1-related myopathies

Abstract Background Pathogenic variants in RYR1 cause a spectrum of rare congenital myopathies associated with intracellular calcium dysregulation. Glutathione redox imbalance has been reported in several Ryr1 disease model systems and clinical studies. NAD+ and NADP are essential cofactors in cellu...

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Main Authors: Tokunbor A. Lawal, Willa Riekhof, Linda Groom, Pooja Varma, Irene C. Chrismer, Angela Kokkinis, Christopher Grunseich, Jessica W. Witherspoon, Muslima S. Razaqyar, Ninet Sinaii, Katherine G. Meilleur, Lichen Xiang, Jana Buzkova, Liliya Euro, Payam Mohassel, Robert T. Dirksen, Joshua J. Todd
Format: Article
Language:English
Published: BMC 2025-08-01
Series:Skeletal Muscle
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Online Access:https://doi.org/10.1186/s13395-025-00390-6
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author Tokunbor A. Lawal
Willa Riekhof
Linda Groom
Pooja Varma
Irene C. Chrismer
Angela Kokkinis
Christopher Grunseich
Jessica W. Witherspoon
Muslima S. Razaqyar
Ninet Sinaii
Katherine G. Meilleur
Lichen Xiang
Jana Buzkova
Liliya Euro
Payam Mohassel
Robert T. Dirksen
Joshua J. Todd
author_facet Tokunbor A. Lawal
Willa Riekhof
Linda Groom
Pooja Varma
Irene C. Chrismer
Angela Kokkinis
Christopher Grunseich
Jessica W. Witherspoon
Muslima S. Razaqyar
Ninet Sinaii
Katherine G. Meilleur
Lichen Xiang
Jana Buzkova
Liliya Euro
Payam Mohassel
Robert T. Dirksen
Joshua J. Todd
author_sort Tokunbor A. Lawal
collection DOAJ
description Abstract Background Pathogenic variants in RYR1 cause a spectrum of rare congenital myopathies associated with intracellular calcium dysregulation. Glutathione redox imbalance has been reported in several Ryr1 disease model systems and clinical studies. NAD+ and NADP are essential cofactors in cellular metabolism and redox homeostasis. NAD+ deficiency has been associated with skeletal muscle bioenergetic deficits in mitochondrial myopathy and sarcopenia. Methods Using a new colorimetric assay and large control dataset (n = 299), we assessed redox balance (glutathione, NAD+, and NADP) in whole blood from 28 RYR1-RM affected individuals (NCT02362425). Analyses were expanded to human skeletal muscle (n = 4), primary myotube cultures (n = 5), and whole blood and skeletal muscle specimens from Ryr1 Y524S mice. The in vitro effects of nicotinamide riboside (NR) on cellular NAD+ content and mitochondrial respirometry were also tested. Results At baseline, a majority of affected individuals exhibited systemic NAD+ deficiency (19/28 [68%] < 21 µM) and increased NADPH concentrations (22/26 [85%] > 1.6 µM). When compared to controls, decreased NAD+/NADH and NADP/NADPH ratios were observed in 9/28 and 23/26 individuals, respectively. In patient-derived myotube cultures (n = 5), NR appeared to increase cellular NAD+ concentrations in a dose and time-dependent manner at 72-h only and favorably modified maximal respiration and ATP production. Average whole blood GSH/GSSG ratio was comparable between groups, and redox imbalance was not observed in Ryr1 Y524S specimens. Conclusions NAD+ and NADP dyshomeostasis was identified in a subset of RYR1-RM affected individuals. Further experiments are warranted to confirm if NAD+ repletion could be an attractive therapeutic approach given the favorable outcomes reported in other neuromuscular disorders. Graphical Abstract
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spelling doaj-art-2d789229ed124c83b25cf2b597a19dba2025-08-24T11:57:55ZengBMCSkeletal Muscle2044-50402025-08-0115111410.1186/s13395-025-00390-6NAD+ dyshomeostasis in RYR1-related myopathiesTokunbor A. Lawal0Willa Riekhof1Linda Groom2Pooja Varma3Irene C. Chrismer4Angela Kokkinis5Christopher Grunseich6Jessica W. Witherspoon7Muslima S. Razaqyar8Ninet Sinaii9Katherine G. Meilleur10Lichen Xiang11Jana Buzkova12Liliya Euro13Payam Mohassel14Robert T. Dirksen15Joshua J. Todd16Skeletal Myopathies Unit, Translational Biobehavioral and Health Disparities Branch, NIH Clinical Center, National Institutes of HealthSkeletal Myopathies Unit, Translational Biobehavioral and Health Disparities Branch, NIH Clinical Center, National Institutes of HealthDepartment of Pharmacology and Physiology, University of Rochester Medical CenterSkeletal Myopathies Unit, Translational Biobehavioral and Health Disparities Branch, NIH Clinical Center, National Institutes of HealthSkeletal Myopathies Unit, Translational Biobehavioral and Health Disparities Branch, NIH Clinical Center, National Institutes of HealthInherited Neuromuscular Diseases Unit, National Institute of Neurological Disorders and Stroke, National Institutes of HealthInherited Neuromuscular Diseases Unit, National Institute of Neurological Disorders and Stroke, National Institutes of HealthSkeletal Myopathies Unit, Translational Biobehavioral and Health Disparities Branch, NIH Clinical Center, National Institutes of HealthSkeletal Myopathies Unit, Translational Biobehavioral and Health Disparities Branch, NIH Clinical Center, National Institutes of HealthBiostatistics and Clinical Epidemiology Service, National Institutes of Health Clinical CenterSkeletal Myopathies Unit, Translational Biobehavioral and Health Disparities Branch, NIH Clinical Center, National Institutes of HealthSkeletal Myopathies Unit, Translational Biobehavioral and Health Disparities Branch, NIH Clinical Center, National Institutes of HealthNADMed LtdNADMed LtdJohns Hopkins University School of MedicineDepartment of Pharmacology and Physiology, University of Rochester Medical CenterNeuromuscular and Neurogenetic Disorders of Childhood Section, National Institute of Neurological Disorders and Stroke, National Institutes of HealthAbstract Background Pathogenic variants in RYR1 cause a spectrum of rare congenital myopathies associated with intracellular calcium dysregulation. Glutathione redox imbalance has been reported in several Ryr1 disease model systems and clinical studies. NAD+ and NADP are essential cofactors in cellular metabolism and redox homeostasis. NAD+ deficiency has been associated with skeletal muscle bioenergetic deficits in mitochondrial myopathy and sarcopenia. Methods Using a new colorimetric assay and large control dataset (n = 299), we assessed redox balance (glutathione, NAD+, and NADP) in whole blood from 28 RYR1-RM affected individuals (NCT02362425). Analyses were expanded to human skeletal muscle (n = 4), primary myotube cultures (n = 5), and whole blood and skeletal muscle specimens from Ryr1 Y524S mice. The in vitro effects of nicotinamide riboside (NR) on cellular NAD+ content and mitochondrial respirometry were also tested. Results At baseline, a majority of affected individuals exhibited systemic NAD+ deficiency (19/28 [68%] < 21 µM) and increased NADPH concentrations (22/26 [85%] > 1.6 µM). When compared to controls, decreased NAD+/NADH and NADP/NADPH ratios were observed in 9/28 and 23/26 individuals, respectively. In patient-derived myotube cultures (n = 5), NR appeared to increase cellular NAD+ concentrations in a dose and time-dependent manner at 72-h only and favorably modified maximal respiration and ATP production. Average whole blood GSH/GSSG ratio was comparable between groups, and redox imbalance was not observed in Ryr1 Y524S specimens. Conclusions NAD+ and NADP dyshomeostasis was identified in a subset of RYR1-RM affected individuals. Further experiments are warranted to confirm if NAD+ repletion could be an attractive therapeutic approach given the favorable outcomes reported in other neuromuscular disorders. Graphical Abstracthttps://doi.org/10.1186/s13395-025-00390-6Congenital myopathyOxidative stressRYR1GlutathioneNAD+NADP
spellingShingle Tokunbor A. Lawal
Willa Riekhof
Linda Groom
Pooja Varma
Irene C. Chrismer
Angela Kokkinis
Christopher Grunseich
Jessica W. Witherspoon
Muslima S. Razaqyar
Ninet Sinaii
Katherine G. Meilleur
Lichen Xiang
Jana Buzkova
Liliya Euro
Payam Mohassel
Robert T. Dirksen
Joshua J. Todd
NAD+ dyshomeostasis in RYR1-related myopathies
Skeletal Muscle
Congenital myopathy
Oxidative stress
RYR1
Glutathione
NAD+
NADP
title NAD+ dyshomeostasis in RYR1-related myopathies
title_full NAD+ dyshomeostasis in RYR1-related myopathies
title_fullStr NAD+ dyshomeostasis in RYR1-related myopathies
title_full_unstemmed NAD+ dyshomeostasis in RYR1-related myopathies
title_short NAD+ dyshomeostasis in RYR1-related myopathies
title_sort nad dyshomeostasis in ryr1 related myopathies
topic Congenital myopathy
Oxidative stress
RYR1
Glutathione
NAD+
NADP
url https://doi.org/10.1186/s13395-025-00390-6
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