The vitamin B3 analogue nicotinamide riboside has only very minor effects on reducing muscle damage in mdx mice

BACKGROUND Vitamin B3 analogue nicotinamide riboside (NR) has been suggested to have beneficial effects on muscle pathology in a mouse model for Duchenne muscular dystrophy (DMD). In muscle dystrophy, NR is thought to act acts by increasing levels of NAD+, to improve mitochondrial functioning and reduce muscle pathology. OBJECTIVE We here aimed to validate the effects of NR to improve muscle quality after eight weeks of treatment in two different mouse models for DMD: the commonly used mdx mouse on a C57BL/10 background (BL10mdx) and the more severely affected mdx mouse on a DBA/2J background (D2-mdx). METHODS To study in more detail whether NR treatment had an impact on muscle pathology, we assessed the expression levels of several markers for DMD pathology (fibrosis, regeneration and inflammation) in diaphragm. RESULTS Our data showed a trend for increase in NAD+-levels in blood; only in the D2-mdx NR-treated mice the NAD+-levels were slightly increased. These markers were elevated in mdx models compared to controls, but not affected by the NR treatment. Histological analysis of muscle tissues indicated a mild treatment effect in D2-mdx mice. CONCLUSIONS Based on our results, testing NR treatment in clinical trials in DMD patients is not warranted.

[1]  S. Lamandé,et al.  Resveratrol Promotes Hypertrophy in Wildtype Skeletal Muscle and Reduces Muscle Necrosis and Gene Expression of Inflammatory Markers in Mdx Mice , 2021, Molecules.

[2]  David W. Hammers,et al.  The D2.mdx mouse as a preclinical model of the skeletal muscle pathology associated with Duchenne muscular dystrophy , 2020, Scientific Reports.

[3]  Chengmei Sun,et al.  Therapeutic Strategies for Duchenne Muscular Dystrophy: An Update , 2020, Genes.

[4]  G. Trifirò,et al.  Global epidemiology of Duchenne muscular dystrophy: an updated systematic review and meta-analysis , 2020, Orphanet Journal of Rare Diseases.

[5]  U. Spitz,et al.  Nicotinamide Riboside—The Current State of Research and Therapeutic Uses , 2020, Nutrients.

[6]  David Frederick,et al.  Complementary NAD+ replacement strategies fail to functionally protect dystrophin-deficient muscle , 2020, Skeletal muscle.

[7]  Mark S. Schmidt,et al.  Niacin Cures Systemic NAD+ Deficiency and Improves Muscle Performance in Adult-Onset Mitochondrial Myopathy. , 2020, Cell metabolism.

[8]  J. Selsby,et al.  Nutraceutical and pharmaceutical cocktails did not preserve diaphragm muscle function or reduce muscle damage in D2‐mdx mice , 2020, Experimental physiology.

[9]  H. Lochmüller,et al.  Life expectancy at birth in Duchenne muscular dystrophy: a systematic review and meta-analysis , 2020, European Journal of Epidemiology.

[10]  David S. Jones,et al.  Understanding the physicochemical properties and degradation kinetics of nicotinamide riboside, a promising vitamin B3nutritional supplement , 2019, Food & nutrition research.

[11]  J. Treebak,et al.  Nicotinamide riboside does not alter mitochondrial respiration, content or morphology in skeletal muscle from obese and insulin‐resistant men , 2019, The Journal of physiology.

[12]  E. Mercuri,et al.  Long-term data with idebenone on respiratory function outcomes in patients with Duchenne muscular dystrophy , 2019, Neuromuscular Disorders.

[13]  J. Selsby,et al.  Nutraceutical and pharmaceutical cocktails did not improve muscle function or reduce histological damage in d2-mdx mice. , 2019, Journal of applied physiology.

[14]  Mark S. Schmidt,et al.  Nicotinamide Riboside Augments the Aged Human Skeletal Muscle NAD+ Metabolome and Induces Transcriptomic and Anti-inflammatory Signatures , 2019, Cell reports.

[15]  H. Knutsen,et al.  Safety of nicotinamide riboside chloride as a novel food pursuant to Regulation (EU) 2015/2283 and bioavailability of nicotinamide from this source, in the context of Directive 2002/46/EC , 2019, EFSA journal. European Food Safety Authority.

[16]  C. Brenner,et al.  Safety and Metabolism of Long-term Administration of NIAGEN (Nicotinamide Riboside Chloride) in a Randomized, Double-Blind, Placebo-controlled Clinical Trial of Healthy Overweight Adults , 2019, Scientific Reports.

[17]  Annemieke Aartsma-Rus,et al.  Natural disease history of the D2-mdx mouse model for Duchenne muscular dystrophy , 2019, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[18]  Y. Horio,et al.  Resveratrol Decreases Oxidative Stress by Restoring Mitophagy and Improves the Pathophysiology of Dystrophin-Deficient mdx Mice , 2018, Oxidative medicine and cellular longevity.

[19]  J. Auwerx,et al.  De novo NAD+ synthesis enhances mitochondrial function and improves health , 2018, Nature.

[20]  K. Davies,et al.  “Of Mice and Measures”: A Project to Improve How We Advance Duchenne Muscular Dystrophy Therapies to the Clinic , 2018, Journal of neuromuscular diseases.

[21]  Mark S. Schmidt,et al.  A randomized placebo-controlled clinical trial of nicotinamide riboside in obese men: safety, insulin-sensitivity, and lipid-mobilizing effects. , 2018, The American journal of clinical nutrition.

[22]  E. White,et al.  Quantitative Analysis of NAD Synthesis-Breakdown Fluxes. , 2018, Cell metabolism.

[23]  V. Mouly,et al.  Combined Therapies for Duchenne Muscular Dystrophy to Optimize Treatment Efficacy , 2018, Front. Genet..

[24]  M. McQueen,et al.  Chronic nicotinamide riboside supplementation is well-tolerated and elevates NAD+ in healthy middle-aged and older adults , 2018, Nature Communications.

[25]  J. Baur,et al.  NAD+ Intermediates: The Biology and Therapeutic Potential of NMN and NR. , 2017, Cell metabolism.

[26]  R. Tian,et al.  An open-label, non-randomized study of the pharmacokinetics of the nutritional supplement nicotinamide riboside (NR) and its effects on blood NAD+ levels in healthy volunteers , 2017, PloS one.

[27]  J. Kleijnen,et al.  The burden, epidemiology, costs and treatment for Duchenne muscular dystrophy: an evidence review , 2017, Orphanet Journal of Rare Diseases.

[28]  J. Baur,et al.  Nicotinamide adenine dinucleotide biosynthesis promotes liver regeneration , 2017, Hepatology.

[29]  J. Auwerx,et al.  NAD+ repletion improves muscle function in muscular dystrophy and counters global PARylation , 2016, Science Translational Medicine.

[30]  M. Mattson,et al.  NAD+ Replenishment Improves Lifespan and Healthspan in Ataxia Telangiectasia Models via Mitophagy and DNA Repair. , 2016, Cell metabolism.

[31]  Mark S. Schmidt,et al.  Nicotinamide riboside is uniquely and orally bioavailable in mice and humans , 2016, Nature Communications.

[32]  B. Gregory,et al.  Loss of NAD Homeostasis Leads to Progressive and Reversible Degeneration of Skeletal Muscle. , 2016, Cell metabolism.

[33]  R. Aebersold,et al.  NAD+ repletion improves mitochondrial and stem cell function and enhances life span in mice , 2016, Science.

[34]  Y. Horio,et al.  SIRT1: A Novel Target for the Treatment of Muscular Dystrophies , 2016, Oxidative medicine and cellular longevity.

[35]  O. Bieri,et al.  Improved Muscle Function in Duchenne Muscular Dystrophy through L-Arginine and Metformin: An Investigator-Initiated, Open-Label, Single-Center, Proof-Of-Concept-Study , 2016, PloS one.

[36]  Andrew D. Williams,et al.  Defects in Mitochondrial ATP Synthesis in Dystrophin-Deficient Mdx Skeletal Muscles May Be Caused by Complex I Insufficiency , 2014, PloS one.

[37]  Barbora Malecova,et al.  STAT3 signaling controls satellite cell expansion and skeletal muscle repair , 2014, Nature Medicine.

[38]  L. Guarente,et al.  Muscle-Specific SIRT1 Gain-of-Function Increases Slow-Twitch Fibers and Ameliorates Pathophysiology in a Mouse Model of Duchenne Muscular Dystrophy , 2014, PLoS genetics.

[39]  E. Schon,et al.  NAD+-Dependent Activation of Sirt1 Corrects the Phenotype in a Mouse Model of Mitochondrial Disease , 2014, Cell metabolism.

[40]  L. Brace,et al.  Defective Mitophagy in XPA via PARP-1 Hyperactivation and NAD+/SIRT1 Reduction , 2014, Cell.

[41]  J. Auwerx,et al.  Effective treatment of mitochondrial myopathy by nicotinamide riboside, a vitamin B3 , 2014, EMBO molecular medicine.

[42]  E. Mercken,et al.  Declining NAD+ Induces a Pseudohypoxic State Disrupting Nuclear-Mitochondrial Communication during Aging , 2013, Cell.

[43]  A. Goldberg,et al.  SIRT1 Protein, by Blocking the Activities of Transcription Factors FoxO1 and FoxO3, Inhibits Muscle Atrophy and Promotes Muscle Growth* , 2013, The Journal of Biological Chemistry.

[44]  L. Guarente,et al.  The NAD+/Sirtuin Pathway Modulates Longevity through Activation of Mitochondrial UPR and FOXO Signaling , 2013, Cell.

[45]  J. Auwerx,et al.  The NAD(+) precursor nicotinamide riboside enhances oxidative metabolism and protects against high-fat diet-induced obesity. , 2012, Cell metabolism.

[46]  C. Ward,et al.  Measuring mitochondrial respiration in intact single muscle fibers. , 2012, American journal of physiology. Regulatory, integrative and comparative physiology.

[47]  O. Dorchies,et al.  Quantitative evaluation of the beneficial effects in the mdx mouse of epigallocatechin gallate, an antioxidant polyphenol from green tea , 2012, Histochemistry and Cell Biology.

[48]  Angeliki Chalkiadaki,et al.  Sirtuins mediate mammalian metabolic responses to nutrient availability , 2012, Nature Reviews Endocrinology.

[49]  S. Messina,et al.  Telomere shortening is associated to TRF1 and PARP1 overexpression in Duchenne muscular dystrophy , 2011, Neurobiology of Aging.

[50]  Yuefang Zhou,et al.  Myosin heavy chain expression in mouse extraocular muscle: more complex than expected. , 2010, Investigative ophthalmology & visual science.

[51]  Hiroshi Yamamoto,et al.  Genetic background affects properties of satellite cells and mdx phenotypes. , 2010, The American journal of pathology.

[52]  P. Puigserver,et al.  AMPK regulates energy expenditure by modulating NAD+ metabolism and SIRT1 activity , 2009, Nature.

[53]  M. Grounds,et al.  Towards developing standard operating procedures for pre-clinical testing in the mdx mouse model of Duchenne muscular dystrophy , 2008, Neurobiology of Disease.

[54]  J. Chamberlain,et al.  Dystrophin‐deficient mdx mice display a reduced life span and are susceptible to spontaneous rhabdomyosarcoma , 2007, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[55]  Jiandie D. Lin,et al.  PGC-1α protects skeletal muscle from atrophy by suppressing FoxO3 action and atrophy-specific gene transcription , 2006, Proceedings of the National Academy of Sciences.

[56]  C. Brenner,et al.  Discoveries of Nicotinamide Riboside as a Nutrient and Conserved NRK Genes Establish a Preiss-Handler Independent Route to NAD+ in Fungi and Humans , 2004, Cell.

[57]  Andrew P. Weir,et al.  Function and genetics of dystrophin and dystrophin-related proteins in muscle. , 2002, Physiological reviews.

[58]  Y. Goto,et al.  Expression of MyoD and myogenin in dystrophic mice, mdx and dy, during regeneration , 2000, Acta Neuropathologica.

[59]  J. Shrager,et al.  The mdx mouse diaphragm reproduces the degenerative changes of Duchenne muscular dystrophy , 1991, Nature.

[60]  E A Barnard,et al.  The molecular basis of muscular dystrophy in the mdx mouse: a point mutation. , 1989, Science.

[61]  A. Monaco,et al.  The complete sequence of dystrophin predicts a rod-shaped cytoskeletal protein , 1988, Cell.

[62]  Eric P. Hoffman,et al.  Dystrophin: The protein product of the duchenne muscular dystrophy locus , 1987, Cell.

[63]  K. Moore,et al.  X chromosome-linked muscular dystrophy (mdx) in the mouse. , 1984, Proceedings of the National Academy of Sciences of the United States of America.

[64]  D. Allen,et al.  Absence of Dystrophin Disrupts Skeletal Muscle Signaling: Roles of Ca2+, Reactive Oxygen Species, and Nitric Oxide in the Development of Muscular Dystrophy. , 2016, Physiological reviews.

[65]  C. Lutz,et al.  Effect of genetic background on the dystrophic phenotype in mdx mice. , 2016, Human molecular genetics.

[66]  B. Spiegelman,et al.  PGC-1alpha regulates the neuromuscular junction program and ameliorates Duchenne muscular dystrophy. , 2007, Genes & development.