bims-moremu 2022-02-13 papers

bims-moremu

Biomed News

on Molecular regulators of muscle mass

Issue of 2022–02–13
37 papers selected by
Anna Vainshtein, Craft Science Inc.

ERK3-MK5 signaling regulates myogenic differentiation and muscle regeneration by promoting FoxO3 degradation.
The Gentle Side of the UPS: Ubiquitin-Proteasome System and the Regulation of the Myogenic Program.
Long Non-coding RNA MALAT1 Is Depleted With Age in Skeletal Muscle in vivo and MALAT1 Silencing Increases Expression of TGF-β1 in vitro.
m6 A demethylase ALKBH5 drives denervation-induced muscle atrophy by targeting HDAC4 to activate FoxO3 signalling.
Human pancreatic tumour organoid-derived factors enhance myogenic differentiation.
DHA alleviates diet-induced skeletal muscle fiber remodeling via FTO/m6A/DDIT4/PGC1α signaling.
Selenoprotein K protects skeletal muscle from damage and is required for satellite cells-mediated myogenic differentiation.
Widespread Mislocalization of FUS Is Associated With Mitochondrial Abnormalities in Skeletal Muscle in Amyotrophic Lateral Sclerosis With FUS Mutations.
The role of the neural stimulus in regulating skeletal muscle hypertrophy.
Myokine-mediated exercise effects: the role of myokine meteorin-like hormone (Metrnl).
Syndecan-4 affects myogenesis via Rac1-mediated actin remodeling and exhibits copy-number amplification and increased expression in human rhabdomyosarcoma tumors.
Low pH up-regulates interleukin-6 mRNA in L6-G8C5 rat skeletal muscle cells independent of pH sensing by SNAT2(SLC38A2) transporters.
Reversal of RNA toxicity in myotonic dystrophy via a decoy RNA-binding protein with high affinity for expanded CUG repeats.
Handling a mature calcium signature through optogenetics improves the differentiation of primary murine myotubes.
Tent5a modulates muscle fiber formation in adolescent idiopathic scoliosis via maintenance of myogenin expression.
Microencapsulated Sertoli cells sustain myoblast proliferation without affecting the myogenic potential. In vitro data.
The effect of hypoxia on myogenic differentiation and multipotency of the skeletal muscle-derived stem cells in mice.
Ryanodine receptor RyR1-mediated elevation of Ca2+ concentration is required for the late stage of myogenic differentiation and fusion.
Intermittent glucocorticoid treatment enhances skeletal muscle performance through sexually dimorphic mechanisms.
Regulation of skeletal myogenesis in C2C12 cells through modulation of Pax7, MyoD, and myogenin via different low-frequency electromagnetic field energies.
Genetic dissection of novel myopathy models reveals a role of CapZα and Leiomodin 3 during myofibril elongation.
Comparison of dystrophin expression following gene editing and gene replacement in an aged preclinical DMD animal model.
Issues on Trainability.
Impaired Proteostasis in Obese Skeletal Muscle Relates to Altered Immunoproteasome Activity.
Physiological and Pathological Relevance of Selective and Nonselective Ca2+ Channels in Skeletal and Cardiac Muscle.
Exercise Capacity Is Improved by Levosimendan in Heart Failure and Sarcopenia via Alleviation of Apoptosis of Skeletal Muscle.
Acute exercise rapidly activates hepatic mitophagic flux.
RhoA/ROCK2 signalling is enhanced by PDGF-AA in fibro-adipogenic progenitor cells: implications for Duchenne muscular dystrophy.
AMPK and the Adaptation to Exercise.
Exploring the role of eRNA in regulating gene expression.
Lean mass sparing in resistance-trained athletes during caloric restriction: the role of resistance training volume.
FNDC5/irisin facilitates muscle-adipose-bone connectivity through ubiquitination-dependent activation of runt-related transcriptional factors RUNX1/2.
Lamina-associated polypeptide 2α is required for intranuclear MRTF-A activity.
Swimming Attenuates Muscle Wasting and Mediates Multiple Signaling Pathways and Metabolites in CT-26 Bearing Mice.
Conformational transitions and ligand-binding to a muscle-type nicotinic acetylcholine receptor.
COVID-19-associated Critical Illness Myopathy with Direct Viral Effects.
Exercise improves angiogenic function of circulating exosomes in type 2 diabetes: Role of exosomal SOD3.

J Cell Physiol. 2022 Feb 09.

ERK3-MK5 signaling regulates myogenic differentiation and muscle regeneration by promoting FoxO3 degradation.

Mathilde Soulez, Pierre-Luc Tanguay, Florence Dô, Junio Dort, Colin Crist, Alexey Kotlyarov, Matthias Gaestel, Mathieu Ferron, Nicolas A Dumont, Sylvain Meloche.

  The physiological functions and downstream effectors of the atypical mitogen-activated protein kinase extracellular signal-regulated kinase 3 (ERK3) remain to be characterized. We recently reported that mice expressing catalytically-inactive ERK3 (Mapk6KD/KD ) exhibit a reduced postnatal growth rate as compared to control mice. Here, we show that genetic inactivation of ERK3 impairs postnatal skeletal muscle growth and adult muscle regeneration after injury. Loss of MAPK-activated protein kinase 5 (MK5) phenocopies the muscle phenotypes of Mapk6KD/KD mice. At the cellular level, genetic or pharmacological inactivation of ERK3 or MK5 induces precocious differentiation of C2C12 or primary myoblasts, concomitant with MyoD activation. Reciprocally, ectopic expression of activated MK5 inhibits myogenic differentiation. Mechanistically, we show that MK5 directly phosphorylates FoxO3, promoting its degradation and reducing its association with MyoD. Depletion of FoxO3 rescues in part the premature differentiation of C2C12 myoblasts observed upon inactivation of ERK3 or MK5. Our findings reveal that ERK3 and its substrate MK5 act in a linear signaling pathway to control postnatal myogenic differentiation.

Keywords:  FoxO; MAP kinases; genetically-engineered mouse models; myogenic differentiation; signal transduction

DOI:  https://doi.org/10.1002/jcp.30695
Front Cell Dev Biol. 2021 ;9 821839

The Gentle Side of the UPS: Ubiquitin-Proteasome System and the Regulation of the Myogenic Program.

Hugo C Olguín.

  In recent years, the ubiquitin-proteasome system (UPS) has emerged as an important regulator of stem cell function. Here we review recent findings indicating that UPS also plays critical roles in the biology of satellite cells, the muscle stem cell responsible for its maintenance and regeneration. While we focus our attention on the control of key transcriptional regulators of satellite cell function, we briefly discuss early studies suggesting the UPS participates more broadly in the regulation of satellite cell stemness and regenerative capacity.

Keywords:  adult stem (AS) cells; myogenesis; myogenic program; proteasome; satellite cells; skeletal muscle; stem cell fate; ubiquitin (Ub)

DOI:  https://doi.org/10.3389/fcell.2021.821839
Front Physiol. 2021 ;12 742004

Long Non-coding RNA MALAT1 Is Depleted With Age in Skeletal Muscle in vivo and MALAT1 Silencing Increases Expression of TGF-β1 in vitro.

Ling Ruan, Bharati Mendhe, Emily Parker, Andrew Kent, Carlos M Isales, William D Hill, Meghan McGee-Lawrence, Sadanand Fulzele, Mark W Hamrick.

  Long non-coding RNAs (lncRNAs) are thought to function as "sponges" for microRNAs, but a role for such competing endogenous RNAs (ceRNAs) in muscle aging is not well understood. We therefore examined in skeletal muscles of young (4-6 months) and aged (22-24) male and female mice the expression of lncRNA MALAT1, which is predicted in silico to bind the senescence-associated microRNA miR-34a-5p. Results indicate a significant decrease in lncRNA MALAT1 expression in mouse skeletal muscle with age that coincides with an age-related increase in miR-34a-5p expression. In vitro studies using mouse C2C12 myoblasts demonstrate that MALAT1 silencing using siRNA increases miR-34a expression, consistent with a role for MALAT1 as an inhibitor of miR-34a-5p activity. Levels of reactive oxygen species (ROS) are known to increase in muscle with age, and so we treated C2C12 cells with hydrogen peroxide (10 and 100 μM) to examine changes in MALAT1 expression. MALAT1 expression decreased significantly with H2O2 treatment, but this effect was attenuated with p53 siRNA. Finally, miR-34a-5p is implicated in tissue fibrosis, and so we assessed the expression of TGF-β1 after MALAT1 silencing. MALAT1 siRNA significantly increased the expression of TGF-β1 in C2C12 cells. These findings suggest that age-related fibrosis and muscle atrophy mediated by ROS may result at least in part from an increase in miR-34a bioavailability resulting from a decline in miR-34a "sponging" due to ceRNA MALAT1 depletion. Crosstalk between MALAT1 and miR-34a may therefore represent a therapeutic target for improving muscle function with aging.

Keywords:  fibrosis; oxidative stress; sarcopenia; senescence; siRNA

DOI:  https://doi.org/10.3389/fphys.2021.742004
J Cachexia Sarcopenia Muscle. 2022 Feb 09.

m6 A demethylase ALKBH5 drives denervation-induced muscle atrophy by targeting HDAC4 to activate FoxO3 signalling.

Yuantong Liu, Tianjian Zhou, Qinghe Wang, Runhan Fu, Zengfu Zhang, Nandi Chen, Zhizhong Li, Guoyong Gao, Songlin Peng, Dazhi Yang.

   BACKGROUND: Skeletal muscle atrophy is a common clinical manifestation of various neurotrauma and neurological diseases. In addition to the treatment of primary neuropathies, it is a clinical condition that should be investigated. FoxO3 activation is an indispensable mechanism in denervation-induced muscle atrophy; however, upstream factors that control FoxO3 expression and activity have not been fully elucidated. N6 -methyladenosine (m6 A) methylation is a novel mode of epitranscriptional gene regulation that affects several cellular processes. However, the biological significance of m6 A modification in FoxO3-dependent atrophy is unknown.
METHODS: We performed gain-of-function and loss-of-function experiments and used denervation-induced muscle atrophy mouse model to evaluate the effects of m6 A modification on muscle mass control and FoxO3 activation. m6 A-sequencing and mass spectrometry analyses were used to establish whether histone deacetylase 4 (HDAC4) is a mediator of m6 A demethylase ALKBH5 regulation of FoxO3. A series of cellular and molecular biological experiments (western blot, immunoprecipitation, half-life assay, m6 A-MeRIP-qPCR, and luciferase reporter assays among others) were performed to investigate regulatory relationships among ALKBH5, HDAC4, and FoxO3.
RESULTS: In skeletal muscles, denervation was associated with a 20.7-31.9% decrease in m6 A levels (P < 0.01) and a 35.6-115.2% increase in demethylase ALKBH5 protein levels (P < 0.05). Overexpressed ALKBH5 reduced m6 A levels, activated FoxO3 signalling, and induced excess loss in muscle wet weight (-10.3% for innervation and -11.4% for denervation, P < 0.05) as well as a decrease in myofibre cross-sectional areas (-35.8% for innervation and -33.3% for denervation, P < 0.05) during innervation and denervation. Specific deletion of Alkbh5 in the skeletal muscles prevented FoxO3 activation and protected mice from denervation-induced muscle atrophy, as evidenced by increased muscle mass (+16.0%, P < 0.05), size (+50.0%, P < 0.05) and MyHC expression (+32.6%, P < 0.05). Mechanistically, HDAC4 was established to be a crucial central mediator for ALKBH5 in enhancing FoxO3 signalling in denervated muscles. ALKBH5 demethylates and stabilizes Hdac4 mRNA. HDAC4 interacts with and deacetylates FoxO3, resulting in a significant increase in FoxO3 expression (+61.3-82.5%, P < 0.01) and activity (+51.6-122.0%, P < 0.001).
CONCLUSIONS: Our findings elucidate on the roles and mechanisms of ALKBH5-mediated m6 A demethylation in the control of muscle mass during denervation and activation of FoxO3 signalling by targeting HDAC4. These results suggest that ALKBH5 is a potential therapeutic target for neurogenic muscle atrophy.

Keywords:  ALKBH5; Denervation; FoxO3; HDAC4; Muscle atrophy; m6A modification

DOI:  https://doi.org/10.1002/jcsm.12929
J Cachexia Sarcopenia Muscle. 2022 Feb 11.

Human pancreatic tumour organoid-derived factors enhance myogenic differentiation.

Rianne D W Vaes, David P J van Dijk, Elham Aïda Farshadi, Steven W M Olde Damink, Sander S Rensen, Ramon C Langen.

   BACKGROUND: Most patients with pancreatic cancer develop cachexia, which is characterized by progressive muscle loss. The mechanisms underlying muscle loss in cancer cachexia remain elusive. Pancreatic tumour organoids are 3D cell culture models that retain key characteristics of the parent tumour. We aimed to investigate the effect of pancreatic tumour organoid-derived factors on processes that determine skeletal muscle mass, including the regulation of muscle protein turnover and myogenesis.
METHODS: Conditioned medium (CM) was collected from human pancreatic cancer cell lines (PK-45H, PANC-1, PK-1, and KLM-1), pancreatic tumour organoid cultures from a severely cachectic (PANCO-9a) and a non-cachectic patient (PANCO-12a), and a normal pancreas organoid culture. Differentiating C2C12 myoblasts and mature C2C12 myotubes were exposed to CM for 24 h or maintained in control medium. In myotubes, NF-kB activation was monitored using a NF-κB luciferase reporter construct, and mRNA expression of E3-ubiquitin ligases and REDD1 was analysed by RT-qPCR. C2C12 myoblast proliferation and differentiation were monitored by live cell imaging and myogenic markers and myosin heavy chain (MyHC) isoforms were assessed by RT-qPCR.
RESULTS: Whereas CM from PK-1 and KLM-1 cells significantly induced NF-κB activation in C2C12 myotubes (PK-1: 3.1-fold, P < 0.001; KLM-1: 2.1-fold, P = 0.01), Atrogin-1/MAFbx and MuRF1 mRNA were only minimally and inconsistently upregulated by the CM of pancreatic cancer cell lines. Similarly, E3-ubiquitin ligases and REDD1 mRNA expression in myotubes were not altered by exposure to pancreatic tumour organoid CM. Compared with the control condition, CM from both PANCO-9a and PANCO-12a tumour organoids increased proliferation of myoblasts, which was accompanied by significant downregulation of the satellite cell marker paired-box 7 (PAX7) (PANCO-9a: -2.1-fold, P < 0.001; PANCO-12a: -2.0-fold, P < 0.001) and myogenic factor 5 (MYF5) (PANCO-9a: -2.1-fold, P < 0.001; PANCO-12a: -1.8-fold, P < 0.001) after 48 h of differentiation. Live cell imaging revealed accelerated alignment and fusion of myoblasts exposed to CM from PANCO-9a and PANCO-12a, which was in line with significantly increased Myomaker mRNA expression levels (PANCO-9a: 2.4-fold, P = 0.001; PANCO-12a: 2.2-fold, P = 0.004). These morphological and transcriptional alterations were accompanied by increased expression of muscle differentiation markers such as MyHC-IIB (PANCO-9a: 2.5-fold, P = 0.04; PANCO-12a: 3.1-fold, P = 0.006). Although the impact of organoid CM on myogenesis was not associated with the cachexia phenotype of the donor patients, it was specific for tumour organoids, as CM of control pancreas organoids did not modulate myogenic fusion.
CONCLUSIONS: These data show that pancreatic tumour organoid-derived factors alter the kinetics of myogenesis, which may eventually contribute to impaired muscle mass maintenance in cancer cachexia.

Keywords:  Cachexia; E3 ubiquitin ligases; Myogenesis; Organoids; Skeletal muscle atrophy

DOI:  https://doi.org/10.1002/jcsm.12917
BMC Biol. 2022 Feb 08. 20(1): 39

DHA alleviates diet-induced skeletal muscle fiber remodeling via FTO/m6A/DDIT4/PGC1α signaling.

Wei Chen, Yushi Chen, Ruifan Wu, Guanqun Guo, Youhua Liu, Botao Zeng, Xing Liao, Yizhen Wang, Xinxia Wang.

   BACKGROUND: Obesity leads to a decline in the exercise capacity of skeletal muscle, thereby reducing mobility and promoting obesity-associated health risks. Dietary intervention has been shown to be an important measure to regulate skeletal muscle function, and previous studies have demonstrated the beneficial effects of docosahexaenoic acid (DHA; 22:6 ω-3) on skeletal muscle function. At the molecular level, DHA and its metabolites were shown to be extensively involved in regulating epigenetic modifications, including DNA methylation, histone modifications, and small non-coding microRNAs. However, whether and how epigenetic modification of mRNA such as N6-methyladenosine (m6A) mediates DHA regulation of skeletal muscle function remains unknown. Here, we analyze the regulatory effect of DHA on skeletal muscle function and explore the involvement of m6A mRNA modifications in mediating such regulation.
RESULTS: DHA supplement prevented HFD-induced decline in exercise capacity and conversion of muscle fiber types from slow to fast in mice. DHA-treated myoblasts display increased mitochondrial biogenesis, while slow muscle fiber formation was promoted through DHA-induced expression of PGC1α. Further analysis of the associated molecular mechanism revealed that DHA enhanced expression of the fat mass and obesity-associated gene (FTO), leading to reduced m6A levels of DNA damage-induced transcript 4 (Ddit4). Ddit4 mRNA with lower m6A marks could not be recognized and bound by the cytoplasmic m6A reader YTH domain family 2 (YTHDF2), thereby blocking the decay of Ddit4 mRNA. Accumulated Ddit4 mRNA levels accelerated its protein translation, and the consequential increased DDIT4 protein abundance promoted the expression of PGC1α, which finally elevated mitochondria biogenesis and slow muscle fiber formation.
CONCLUSIONS: DHA promotes mitochondrial biogenesis and skeletal muscle fiber remodeling via FTO/m6A/DDIT4/PGC1α signaling, protecting against obesity-induced decline in skeletal muscle function.

Keywords:  DHA; FTO; Muscle fiber; Obesity; PGC1α

DOI:  https://doi.org/10.1186/s12915-022-01239-w
Redox Biol. 2022 Feb 04. pii: S2213-2317(22)00027-1. [Epub ahead of print]50 102255

Selenoprotein K protects skeletal muscle from damage and is required for satellite cells-mediated myogenic differentiation.

Shengchen Wang, Xia Zhao, Qingqing Liu, Yue Wang, Shu Li, Shiwen Xu.

  The regeneration of adult skeletal muscle after injury is primarily initiated by satellite cells (SCs), but the regulatory mechanisms of cells committed to myogenic differentiation remain poorly explored. Small molecular selenoprotein K (SelK) plays crucial roles in the modulation of endoplasmic reticulum (ER) stress and against oxidative stress. Here, we first showed that SelK expression is activated in myogenic cells during differentiation both in vivo and in vitro. Meanwhile, loss of SelK delayed skeletal muscle regeneration, inhibited the development of myoblasts into myotubes, and was accompanied by reduced expression of myogenic regulatory factors (MRFs). Moreover, ER stress, intracellular reactive oxygen species (ROS), autophagy and apoptosis under myogenesis induction were more severe in SelK-deficient mice and cells than in the corresponding control groups. Supplementation with specific inhibitors to alleviate excessive ER stress or oxidative stress partly rescued the differentiation potential and formation of myotubes. Notably, we demonstrated that Self-mediated regulation of cellular redox status was primarily derived from its subsequent effects on ER stress. Together, our results suggest that SelK protects skeletal muscle from damage and is a crucial regulator of myogenesis.

Keywords:  Endoplasmic reticulum stress; Myogenesis; Oxidative stress; Satellite cells; Selenoprotein K; Skeletal muscle

DOI:  https://doi.org/10.1016/j.redox.2022.102255
J Neuropathol Exp Neurol. 2022 Feb 09. pii: nlac004. [Epub ahead of print]

Widespread Mislocalization of FUS Is Associated With Mitochondrial Abnormalities in Skeletal Muscle in Amyotrophic Lateral Sclerosis With FUS Mutations.

Meng Yu, Xutong Zhao, Wei Wu, Qingqing Wang, Jing Liu, Wei Zhang, Yun Yuan, Daojun Hong, Zhaoxia Wang, Jianwen Deng.

  Mutations in the fused in sarcoma (FUS) gene have been reported to be the most common genetic cause of early-onset amyotrophic lateral sclerosis (ALS); cytoplasmic inclusions containing FUS protein are the predominant pathological feature. Recent studies indicated that mutant FUS impaired neuromuscular junctions and induced muscle intrinsic toxicity in cell and animal models. However, the role of FUS in muscle degeneration remains unclear. In this study, we investigated FUS protein distribution in skeletal muscle fibers in ALS-FUS. Our data show that cytoplasmic mislocalized FUS in the unaggregated form represented a remarkable pathological feature in affected muscle fibers in ALS-FUS. Additional studies found that cytoplasmic FUS colocalized with some mitochondria and was associated with mitochondrial swelling and disorganized cristae. RNA sequencing and quantitative real-time polymerase chain reaction analyses indicated downregulation of the key subunits of mitochondrial oxidative phosphorylation complexes in the affected skeletal muscle in ALS-FUS patients. Further immunoblot analysis showed increased levels of FUS, but decreased levels of Cox I (subunit of complex IV) in ALS-FUS patients compared with age-matched controls. This is the first demonstration of the close association of cytoplasmic mislocalized FUS with mitochondrial dysfunction in skeletal muscle, implicating the presence of a cell-autonomous mechanism in muscle degeneration in ALS.

Keywords:  Amyotrophic lateral sclerosis (ALS); Cell autonomous; Fused in sarcoma (FUS); Mislocalization; Mitochondrial damage; Oxidative phosphorylation; Skeletal muscle

DOI:  https://doi.org/10.1093/jnen/nlac004
Eur J Appl Physiol. 2022 Feb 09.

The role of the neural stimulus in regulating skeletal muscle hypertrophy.

Carlos Alix-Fages, Alessandro Del Vecchio, Eneko Baz-Valle, Jordan Santos-Concejero, Carlos Balsalobre-Fernández.

  Resistance training is frequently performed with the goal of stimulating muscle hypertrophy. Due to the key roles motor unit recruitment and mechanical tension play to induce muscle growth, when programming, the manipulation of the training variables is oriented to provoke the correct stimulus. Although it is known that the nervous system is responsible for the control of motor units and active muscle force, muscle hypertrophy researchers and trainers tend to only focus on the adaptations of the musculotendinous unit and not in the nervous system behaviour. To better guide resistance exercise prescription for muscle hypertrophy and aiming to delve into the mechanisms that maximize this goal, this review provides evidence-based considerations for possible effects of neural behaviour on muscle growth when programming resistance training, and future neurophysiological measurement that should be tested when training to increase muscle mass. Combined information from the neural and muscular structures will allow to understand the exact adaptations of the muscle in response to a given input (neural drive to the muscle). Changes at different levels of the nervous system will affect the control of motor units and mechanical forces during resistance training, thus impacting the potential hypertrophic adaptations. Additionally, this article addresses how neural adaptations and fatigue accumulation that occur when resistance training may influence the hypertrophic response and propose neurophysiological assessments that may improve our understanding of resistance training variables that impact on muscular adaptations.

Keywords:  Fatigue; Muscle growth; Neural adaptations; Neuroscience; Resistance training

DOI:  https://doi.org/10.1007/s00421-022-04906-6
Growth Factors. 2022 Feb 08. 1-8

Myokine-mediated exercise effects: the role of myokine meteorin-like hormone (Metrnl).

Hamid Alizadeh.

  Meterorin-like hormone (Metrnl), as a novel secreted factor, has been shown to be involved in physiological and pathophysiological processes. The behaviour of Metrnl in metabolic conditions like type 2 diabetes is conflicting. Metrnl-mediated (treatment with Metrnl) auto/paracrine actions in skeletal muscle are glucose uptake, fat oxidation and muscle regeneration. Exercise-induced Metrnl actions are increased fat oxidation in both skeletal muscle and adipose tissue, the control of inflammation in adipose tissue (metainflammation), and the regulation of muscle regeneration. Based on the current knowledge, Metrnl as a myokine can establish the muscle-fat crosstalk; however, the ability of Metrnl as a myokine to create other crosstalks remains unclear yet. Additionally, given the considerable anti-inflammatory roles of Metrnl in muscle regeneration, it could be a potential therapeutic candidate for muscle-related inflammatory diseases and ageing skeletal muscle which need to be addressed in the future studies.

Keywords:  Metrnl as a myokine; Metrnl-driven signalling pathways; circulating Metrnl level; metabolic health

DOI:  https://doi.org/10.1080/08977194.2022.2032689
Cell Mol Life Sci. 2022 Feb 07. 79(2): 122

Syndecan-4 affects myogenesis via Rac1-mediated actin remodeling and exhibits copy-number amplification and increased expression in human rhabdomyosarcoma tumors.

Kitti Szabo, Daniel Varga, Attila Gergely Vegh, Ning Liu, Xue Xiao, Lin Xu, Laszlo Dux, Miklos Erdelyi, Laszlo Rovo, Aniko Keller-Pinter.

  Skeletal muscle demonstrates a high degree of regenerative capacity repeating the embryonic myogenic program under strict control. Rhabdomyosarcoma is the most common sarcoma in childhood and is characterized by impaired muscle differentiation. In this study, we observed that silencing the expression of syndecan-4, the ubiquitously expressed transmembrane heparan sulfate proteoglycan, significantly enhanced myoblast differentiation, and fusion. During muscle differentiation, the gradually decreasing expression of syndecan-4 allows the activation of Rac1, thereby mediating myoblast fusion. Single-molecule localized superresolution direct stochastic optical reconstruction microscopy (dSTORM) imaging revealed nanoscale changes in actin cytoskeletal architecture, and atomic force microscopy showed reduced elasticity of syndecan-4-knockdown cells during fusion. Syndecan-4 copy-number amplification was observed in 28% of human fusion-negative rhabdomyosarcoma tumors and was accompanied by increased syndecan-4 expression based on RNA sequencing data. Our study suggests that syndecan-4 can serve as a tumor driver gene in promoting rabdomyosarcoma tumor development. Our results contribute to the understanding of the role of syndecan-4 in skeletal muscle development, regeneration, and tumorigenesis.

Keywords:  Actin; Atomic force microscopy; Muscle differentiation; Myoblast fusion; Proteoglycan; Rac1; Rhabdomyosarcoma; Syndecan-4; dSTORM superresolution microscopy

DOI:  https://doi.org/10.1007/s00018-021-04121-0
FASEB Bioadv. 2022 Feb;4(2): 138-152

Low pH up-regulates interleukin-6 mRNA in L6-G8C5 rat skeletal muscle cells independent of pH sensing by SNAT2(SLC38A2) transporters.

Ziyad Aldosari, Nima Abbasian, Katherine Robinson, Alan Bevington, Emma Watson.

  Exercise is known to create a transient, but potent increase in skeletal muscle expression of potentially anti-inflammatory myokine interleukin-6 (IL-6). This effect may be clinically important in managing chronic inflammatory states. It has previously been proposed that lactic acidosis following exercise promotes this IL-6 up-regulation, but the mechanism of this acidosis effect is unknown. Rat skeletal muscle cell line L6-G8C5 has been used previously to model metabolic effects of acidosis, sensing low pH through the resulting inhibition of amino acid transporter SNAT2(SLC38A2). Use of ionophore ionomycin to model the rise in intracellular Ca2+ concentration occurring in contracting muscle strongly up-regulates IL-6 mRNA in L6-G8C5 myotubes. This study used this model to test the hypothesis that low extracellular pH (7.1) enhances ionomycin-induced IL-6 mRNA up-regulation by inhibiting SNAT2. Incubation of L6-G8C5 myotubes for 6 h with 0.5 µM ionomycin at control pH (7.4) resulted in a 15-fold increase in IL-6 mRNA which was further enhanced (1.74-fold) at pH 7.1. In contrast low pH had no significant effect on IL-6 mRNA without ionomycin, nor on the IL-6 mRNA increase that was induced by cyclic stretch. Even though pH 7.1 halved the transport activity of SNAT2, alternative methods of SNAT2 inhibition (JNK inhibitor SP600125; SNAT2 antagonist MeAIB; or SNAT2 silencing with siRNA) did not mimic the enhancing effect of low pH on IL-6 mRNA. On the contrary, JNK inhibition blunted the effect of pH 7.1 with ionomycin, but had no effect at pH 7.4. It is concluded that low pH promotes Ca2+/ionomycin-induced up-regulation of IL-6 mRNA through a novel SNAT2-independent JNK-dependent pH-sensing pathway not previously described in this skeletal muscle model.

Keywords:  JNK; SLC38A2; SNAT2; interleukin‐6 mRNA; pH; skeletal muscle

DOI:  https://doi.org/10.1096/fba.2021-00088
Nat Biomed Eng. 2022 Feb 10.

Reversal of RNA toxicity in myotonic dystrophy via a decoy RNA-binding protein with high affinity for expanded CUG repeats.

Ludovic Arandel, Magdalena Matloka, Arnaud F Klein, Frédérique Rau, Alain Sureau, Michel Ney, Aurélien Cordier, Maria Kondili, Micaela Polay-Espinoza, Naira Naouar, Arnaud Ferry, Mégane Lemaitre, Séverine Begard, Morvane Colin, Chloé Lamarre, Hélène Tran, Luc Buée, Joëlle Marie, Nicolas Sergeant, Denis Furling.

Myotonic dystrophy type 1 (DM1) is an RNA-dominant disease whose pathogenesis stems from the functional loss of muscleblind-like RNA-binding proteins (RBPs), which causes the formation of alternative-splicing defects. The loss of functional muscleblind-like protein 1 (MBNL1) results from its nuclear sequestration by mutant transcripts containing pathogenic expanded CUG repeats (CUGexp). Here we show that an RBP engineered to act as a decoy for CUGexp reverses the toxicity of the mutant transcripts. In vitro, the binding of the RBP decoy to CUGexp in immortalized muscle cells derived from a patient with DM1 released sequestered endogenous MBNL1 from nuclear RNA foci, restored MBNL1 activity, and corrected the transcriptomic signature of DM1. In mice with DM1, the local or systemic delivery of the RBP decoy via an adeno-associated virus into the animals' skeletal muscle led to the long-lasting correction of the splicing defects and to ameliorated disease pathology. Our findings support the development of decoy RBPs with high binding affinities for expanded RNA repeats as a therapeutic strategy for myotonic dystrophies.

DOI: https://doi.org/10.1038/s41551-021-00838-2
Cell Calcium. 2022 Feb 03. pii: S0143-4160(22)00021-5. [Epub ahead of print]103 102546

Handling a mature calcium signature through optogenetics improves the differentiation of primary murine myotubes.

Charles-Albert Chapotte-Baldacci, Christian Cognard, Patrick Bois, Aurélien Chatelier, Stéphane Sebille.

  Calcium takes part in numerous cellular processes such as proliferation, migration, differentiation, or cell death and plays a particular role in myogenesis of skeletal muscle. Indeed, intracellular calcium signaling participates, in a non-negligeable manner, to the "on" signal of muscle differentiation from undifferentiated cells to differentiated myotubes. Therefore, this differentiation can be modulated by controlling calcium activity with electrical or optogenetic stimulation approaches. In this study, we used the optogenetic tool channelrhodopsin 2 (ChR2) to control calcium activity and to modulate skeletal muscle differentiation. Using primary cultures of mouse myotubes, we showed that ChR2 stimulation was well-adapted to control intracellular calcium activity at the single cell or whole culture scale. To modulate the calcium-dependent myotube differentiation, we used an optical stimulation protocol based on GCAMP6s-decoded spontaneous calcium activity patterns of differentiated myotubes. The optical training of myotubes increased the fusion index and their contractile ability. This study demonstrates that handling a mature calcium signature with such optogenetic tool improves the differentiation of primary murine myotubes.

Keywords:  Calcium homeostasis; ChR2; GCaMP; Myogenesis; Optogenetics; Primary culture

DOI:  https://doi.org/10.1016/j.ceca.2022.102546
Cell Prolif. 2022 Feb 09. e13183

Tent5a modulates muscle fiber formation in adolescent idiopathic scoliosis via maintenance of myogenin expression.

Ming Luo, Huiliang Yang, Diwei Wu, Xuanhe You, Shishu Huang, Yueming Song.

   OBJECTIVE: Paravertebral muscle asymmetry may be involved in the pathogenesis of adolescent idiopathic scoliosis (AIS), and the Tent5a protein was recently identified as a novel active noncanonical poly(A) polymerase. We, therefore, explored the function of the AIS susceptibility gene Tent5a in myoblasts.
MATERIALS AND METHODS: RNA-seq of AIS paravertebral muscle was performed, and the molecular differences in paravertebral muscle were investigated. Twenty-four AIS susceptibility genes were screened, and differential expression of Tent5a in paravertebral muscles was confirmed with qPCR and Western blot. After the knockdown of Tent5a, the functional effects of Tent5a on C2C12 cell proliferation, migration, and apoptosis were detected by Cell Counting Kit-8 assay, wound-healing assay, and TUNEL assay, respectively. Myogenic differentiation markers were tested with immunofluorescence and qPCR in vitro, and muscle fiber formation was compared in vivo.
RESULTS: The AIS susceptibility gene Tent5a was differentially expressed in AIS paravertebral muscles. Tent5a knockdown inhibited the proliferation and migration of C2C12 cells and inhibited the maturation of type I muscle fibers in vitro and in vivo. Mechanistically, the expression of myogenin was decreased along with the suppression of Tent5a.
CONCLUSIONS: Tent5a plays an important role in the proliferation and migration of myoblasts, and it regulates muscle fiber maturation by maintaining the stability of myogenin. Tent5a may be involved in the pathogenesis of AIS by regulating the formation of muscle fiber type I.

Keywords:   Tent5a ; adolescent idiopathic scoliosis; myoblast differentiation; myogenin; paravertebral muscle asymmetry

DOI:  https://doi.org/10.1111/cpr.13183
Data Brief. 2022 Feb;40 107744

Microencapsulated Sertoli cells sustain myoblast proliferation without affecting the myogenic potential. In vitro data.

Sara Chiappalupi, Laura Salvadori, Francesca Mancuso, Iva Arato, Mario Calvitti, Francesca Riuzzi, Riccardo Calafiore, Giovanni Luca, Guglielmo Sorci.

  Sertoli cells (SeC) isolated from porcine testes have shown direct effects on muscle precursor cells sustaining C2C12 myoblasts proliferation and inhibiting oxidative stress and apoptosis in the early phase of the differentiation process, and stimulating myoblast fusion into myotubes and the expression of markers of myogenic differentiation in the late phase. This suggested that the cocktail of factors secreted by SeC stimulates proliferation in myoblasts without weakening their myogenic potential resulting in the formation of the critical myoblast amount necessary to rebuild the required muscle mass upon a damage. Here, we show that co-culturing C2C12 myoblasts with high doses of SeC microencapsulated in clinical grade alginate-based microcapsules (MC-SeC) for three days in differentiation medium (DM) translates into increased cell numbers and almost absence of myotube formation. However, after removal of MC-SeC, an intense fusion activity into myotubes was observed culminating in a fusion index similar to that of control after additional three days of culture in DM. These data definitely demonstrate that SeC-derived factors preserve the myogenic potential while sustaining cell proliferation in C2C12 myoblasts.

Keywords:  ALG, sodium alginate; AMH, anti-Müllerian hormone; ASMA, alpha-smooth muscle actin; BSA, bovine serum albumin; DAPI, 4′,6-diamidino-2-phenylindole; DM, differentiation medium; E-MC, empty microcapsules; EB, ethidium bromide; EDTA, ethylene-diamine tetra-acetic acid; FBS, foetal bovine serum; FDA, fluorescein diacetate; Fusion index; HBSS, Hanks’ balanced salt solution; HG-DMEM, high-glucose Dulbecco’s modified Eagle’s medium; HS, horse serum; INSL3, insulin-like3; ITS, insulin-transferrin-selenium; MC-SeC, microencapsulated Sertoli cells; Microencapsulation; Myoblast; Myoblast proliferation; Myogenic potential; P/S, penicillin/streptomycin; PFA, paraformaldehyde; PGP9.5, protein gene product 9.5; SeC, Sertoli cells; Sertoli cell; TRIS, tris(hydroxymethyl)aminomethane

DOI:  https://doi.org/10.1016/j.dib.2021.107744
Stem Cell Res Ther. 2022 Feb 05. 13(1): 56

The effect of hypoxia on myogenic differentiation and multipotency of the skeletal muscle-derived stem cells in mice.

Mohamed I Elashry, Mebrie Kinde, Michele C Klymiuk, Asmaa Eldaey, Sabine Wenisch, Stefan Arnhold.

   BACKGROUND: Skeletal muscle-derived stem cells (SC) have become a promising approach for investigating myogenic differentiation and optimizing tissue regeneration. Muscle regeneration is performed by SC, a self-renewal cell population underlying the basal lamina of muscle fibers. Here, we examined the impact of hypoxia condition on the regenerative capacity of SC either in their native microenvironment or via isolation in a monolayer culture using ectopic differentiation inductions. Furthermore, the effect of low oxygen tension on myogenic differentiation protocols of the myoblasts cell line C2C12 was examined.
METHODS: Hind limb muscles of wild type mice were processed for both SC/fiber isolation and myoblast extraction using magnetic beads. SC were induced for myogenic, adipogenic and osteogenic commitments under normoxic (21% O2) and hypoxic (3% O2) conditions. SC proliferation and differentiation were evaluated using histological staining, immunohistochemistry, morphometric analysis and RT-qPCR. The data were statistically analyzed using ANOVA.
RESULTS: The data revealed enhanced SC proliferation and motility following differentiation induction after 48 h under hypoxia. Following myogenic induction, the number of undifferentiated cells positive for Pax7 were increased at 72 h under hypoxia. Hypoxia upregulated MyoD and downregulated Myogenin expression at day-7 post-myogenic induction. Hypoxia promoted both SC adipogenesis and osteogenesis under respective induction as shown by using Oil Red O and Alizarin Red S staining. The expression of adipogenic markers; peroxisome proliferator activated receptor gamma (PPARγ) and fatty acid-binding protein 4 (FABP4) were upregulated under hypoxia up to day 14 compared to normoxic condition. Enhanced osteogenic differentiation was detected under hypoxic condition via upregulation of osteocalcin and osteopontin expression up to day 14 as well as, increased calcium deposition at day 21. Hypoxia exposure increases the number of adipocytes and the size of fat vacuoles per adipocyte compared to normoxic culture. Combining the differentiation medium with dexamethasone under hypoxia improves the efficiency of the myogenic differentiation protocol of C2C12 by increasing the length of the myotubes.
CONCLUSIONS: Hypoxia exposure increases cell resources for clinical applications and promotes SC multipotency and thus beneficial for tissue regeneration.

Keywords:  Differentiation; Hypoxia; Multipotency; Muscle Stem cells; Proliferation

DOI:  https://doi.org/10.1186/s13287-022-02730-5
J Anim Sci Biotechnol. 2022 Feb 11. 13(1): 9

Ryanodine receptor RyR1-mediated elevation of Ca2+ concentration is required for the late stage of myogenic differentiation and fusion.

Kai Qiu, Yubo Wang, Doudou Xu, Linjuan He, Xin Zhang, Enfa Yan, Lu Wang, Jingdong Yin.

   BACKGROUND: Cytosolic Ca2+ plays vital roles in myogenesis and muscle development. As a major Ca2+ release channel of endoplasmic reticulum (ER), ryanodine receptor 1 (RyR1) key mutations are main causes of severe congenital myopathies. The role of RyR1 in myogenic differentiation has attracted intense research interest but remains unclear.
RESULTS: In the present study, both RyR1-knockdown myoblasts and CRISPR/Cas9-based RyR1-knockout myoblasts were employed to explore the role of RyR1 in myogenic differentiation, myotube formation as well as the potential mechanism of RyR1-related myopathies. We observed that RyR1 expression was dramatically increased during the late stage of myogenic differentiation, accompanied by significantly elevated cytoplasmic Ca2+ concentration. Inhibition of RyR1 by siRNA-mediated knockdown or chemical inhibitor, dantrolene, significantly reduced cytosolic Ca2+ and blocked multinucleated myotube formation. The elevation of cytoplasmic Ca2+ concentration can effectively relieve myogenic differentiation stagnation by RyR1 inhibition, demonstrating that RyR1 modulates myogenic differentiation via regulation of Ca2+ release channel. However, RyR1-knockout-induced Ca2+ leakage led to the severe ER stress and excessive unfolded protein response, and drove myoblasts into apoptosis.
CONCLUSIONS: Therefore, we concluded that Ca2+ release mediated by dramatic increase in RyR1 expression is required for the late stage of myogenic differentiation and fusion. This study contributes to a novel understanding of the role of RyR1 in myogenic differentiation and related congenital myopathies, and provides a potential target for regulation of muscle characteristics and meat quality.

Keywords:  Apoptosis; Ca2+ homeostasis; Endoplasmic reticulum stress; Myoblast fusion; Myogenic differentiation; RyR1 knockout

DOI:  https://doi.org/10.1186/s40104-021-00668-x
J Clin Invest. 2022 Feb 10. pii: e149828. [Epub ahead of print]

Intermittent glucocorticoid treatment enhances skeletal muscle performance through sexually dimorphic mechanisms.

Isabella M Salamone, Mattia Quattrocelli, David Y Barefield, Patrick G Page, Ibrahim Tahtah, Michele Hadhazy, Garima Tomar, Elizabeth M McNally.

  Glucocorticoid steroids are commonly prescribed for many inflammatory conditions, but chronic daily use produces adverse effects including muscle wasting and weakness. In contrast, shorter glucocorticoid pulses may improve athletic performance, although the mechanisms remain unclear. Muscle is sexually dimorphic and comparatively little is known about how male and female muscles respond to glucocorticoid steroids. We investigated the impact of once-weekly glucocorticoid exposure on skeletal muscle performance comparing male and female mice. One month of once-weekly glucocorticoid dosing improved muscle specific force in both males and females. Transcriptomic profiling of isolated myofibers identified a striking sexually dimorphic response to weekly glucocorticoids. Male myofibers had increased expression of genes in the IGF1/PI3K pathway and calcium handling, while female myofibers had profound upregulation of lipid metabolism genes. Muscles from weekly prednisone-treated males had improved calcium handling, while comparably treated female muscles had reduced intramuscular triglycerides. Consistent with altered lipid metabolism, weekly prednisone-treated female mice had greater endurance relative to controls. Using chromatin immunoprecipitation, we defined a sexually dimorphic chromatin landscape after weekly prednisone. These results demonstrate that weekly glucocorticoid exposure elicits distinct pathways in males versus females resulting in enhanced performance.

Keywords:  Calcium; Endocrinology; Insulin signaling; Muscle Biology; Skeletal muscle

DOI:  https://doi.org/10.1172/JCI149828
Technol Health Care. 2022 Jan 25.

Regulation of skeletal myogenesis in C2C12 cells through modulation of Pax7, MyoD, and myogenin via different low-frequency electromagnetic field energies.

Jiaqi Bi, Hong Jing, ChenLiang Zhou, Peng Gao, Fujun Han, Gang Li, Shiwei Zhang.

   BACKGROUND: A low-frequency electromagnetic field (LF-EMF) exerts important biological effects on the human body.
OBJECTIVE: We previously studied the immunity and atrophy of gastrocnemius muscles in rats with spinal cord injuries and found that LF-EMF with a magnetic flux density of 1.5 mT exerted excellent therapeutic and preventive effects on reducing myotubes and increasing spatium intermusculare. However, the effects of LF-EMF on all stages of skeletal myogenesis, such as activation, proliferation, differentiation, and fusion of satellite cells to myotubes as stimulated by myogenic regulatoryfactors (MRFs), have not been fully elucidated.
METHODS: This study investigated the optimal LF-EMF magnetic flux density that exerted maximal effects on all stages of C2C12 cell skeletal myogenesis as well as its impact on regulatory MRFs.
RESULTS: The results showed that an LF-EMF with a magnetic flux density of 2.0 mT could activate C2C12 cells and upregulate the proliferation-promoting transcription factor PAX7. On the other hand, 1.5 mT EMF could upregulate the expression of MyoD and myogenin.
CONCLUSION: LF-EMF could prevent the disappearance of myotubes, with different magnetic flux densities of LF-EMF exerting independent and positive effects on skeletal myogenesis such as satellite cell activation and proliferation, muscle cell differentiation, and myocyte fusion.

Keywords:  C2C12 cells; MyoD; Myogenin; Pax7; low-frequency electromagnetic field (LF-EMF)

DOI:  https://doi.org/10.3233/THC-THC228034
PLoS Genet. 2022 Feb 11. 18(2): e1010066

Genetic dissection of novel myopathy models reveals a role of CapZα and Leiomodin 3 during myofibril elongation.

Joachim Berger, Silke Berger, Yu Shan G Mok, Mei Li, Hakan Tarakci, Peter D Currie.

Myofibrils within skeletal muscle are composed of sarcomeres that generate force by contraction when their myosin-rich thick filaments slide past actin-based thin filaments. Although mutations in components of the sarcomere are a major cause of human disease, the highly complex process of sarcomere assembly is not fully understood. Current models of thin filament assembly highlight a central role for filament capping proteins, which can be divided into three protein families, each ascribed with separate roles in thin filament assembly. CapZ proteins have been shown to bind the Z-disc protein α-actinin to form an anchoring complex for thin filaments and actin polymerisation. Subsequent thin filaments extension dynamics are thought to be facilitated by Leiomodins (Lmods) and thin filament assembly is concluded by Tropomodulins (Tmods) that specifically cap the pointed end of thin filaments. To study thin filament assembly in vivo, single and compound loss-of-function zebrafish mutants within distinct classes of capping proteins were analysed. The generated lmod3- and capza1b-deficient zebrafish exhibited aspects of the pathology caused by variations in their human orthologs. Although loss of the analysed main capping proteins of the skeletal muscle, capza1b, capza1a, lmod3 and tmod4, resulted in sarcomere defects, residual organised sarcomeres were formed within the assessed mutants, indicating that these proteins are not essential for the initial myofibril assembly. Furthermore, detected similarity and location of myofibril defects, apparent at the peripheral ends of myofibres of both Lmod3- and CapZα-deficient mutants, suggest a function in longitudinal myofibril growth for both proteins, which is molecularly distinct to the function of Tmod4.

DOI: https://doi.org/10.1371/journal.pgen.1010066
Mol Ther. 2022 Feb 08. pii: S1525-0016(22)00086-7. [Epub ahead of print]

Comparison of dystrophin expression following gene editing and gene replacement in an aged preclinical DMD animal model.

Niclas E Bengtsson, Julie M Crudele, Jordan M Klaiman, Christine L Halbert, Stephen D Hauschka, Jeffrey S Chamberlain.

Gene-editing has shown promise for correcting or bypassing dystrophin mutations in Duchenne muscular dystrophy (DMD). However, pre-clinical studies have focused on young animals with limited muscle fibrosis and wasting, thereby favoring muscle transduction, myonuclear editing and prevention of disease progression. Here we explore muscle-specific dystrophin-gene editing following intramuscular delivery of AAV6:CK8e-CRISPR/SaCas9 in 3- and 8-year-old dystrophic CXMD dogs, and provide a qualitative comparison to AAV6:CK8e-micro-dystrophin gene-replacement at 6-weeks post-treatment. Gene-editing restored the dystrophin reading-frame in ∼1.3% of genomes and in up to 4.0 % of dystrophin transcripts following excision of a 105 kb mutation-containing region spanning exons 6-8. However, resulting dystrophin expression levels and effects on muscle pathology were greater with the use of micro-dystrophin gene transfer. This study demonstrates that our muscle-specific multi-exon deletion strategy can correct a frequently mutated region of the dystrophin gene in an aged large animal DMD model, but underscores that further enhancements are required to reach efficiencies comparable to AAV-micro-dystrophin. Our observations also indicate that treatment efficacy and state of muscle pathology at the time of intervention are linked, suggesting the need for additional methodological optimizations related to age and disease progression to achieve relevant clinical translation of CRISPR-based therapies to all DMD patients.

DOI: https://doi.org/10.1016/j.ymthe.2022.02.003
Front Physiol. 2021 ;12 790196

Issues on Trainability.

Zsolt Radak, Albert W Taylor.

  Trainability is an adaptive response to given exercise loads and must be localized to the targeted physiological function since exercise-induced acute and chronic adaptations are systemic. Lack of adaptation or moderate level of adaptation in one organ or one physiological function would not mean that other organs or functions would not benefit from exercise training. The most beneficial training load could easily be different for skeletal muscle, brain, the gastro-intestinal track, or the immune systems. Hence, the term of non-responders should be used with caution and just referred to a given organ, cell type, molecular signaling, or function. The present paper aims to highlight some, certainly not all, issues on trainability especially related to muscle and cardiovascular system. The specificity of trainability and the systemic nature of exercise-induced adaptation are discussed, and the paper aims to provide suggestions on how to improve performance when faced with non-responders.

Keywords:  VO2max; non-responders; resistance training; responders; systemic adaptation

DOI:  https://doi.org/10.3389/fphys.2021.790196
Appl Physiol Nutr Metab. 2022 Feb 11.

Impaired Proteostasis in Obese Skeletal Muscle Relates to Altered Immunoproteasome Activity.

Emma Fletcher, Michael Wiggs, K Leigh Greathouse, Grant B Morgan, Paul M Gordon.

Obesity-associated inflammation and/or oxidative stress can damage intramuscular proteins and jeopardize muscle integrity. The immunoproteasome (iProt) is vital to remove oxidatively modified proteins, but this function may be compromised with obesity. We sought to elucidate whether diet-induced obesity (DIO) alters intramuscular iProt content and activity in mice to identify a possible mechanism for impaired muscle proteostasis in the obese state. Total proteasome content and activity and estimates of muscle oxidative damage, inflammation, muscle mass and strength were also assessed. Twenty-three male, 5-week-old C57BL/6J mice were fed a high-fat, high-sucrose (HFS, 45% kcal fat, 17% sucrose, n = 12) or low-fat, low-sucrose (LFS, 10% kcal fat, 0% sucrose, n = 11) diet for 12 weeks. Strength was assessed via a weightlifting test. Despite no change in pro-inflammatory cytokines (P > 0.05), oxidative protein damage was elevated within the gastrocnemius (P = 0.036) and tibialis anterior (P = 0.033) muscles of HFS-fed mice. Intramuscular protein damage coincided with reduced iProt and total proteasome activity (P < 0.05), and reductions in relative muscle mass (P < 0.001). Therefore, proteasome dysregulation occurs in obese muscle and may be a critical link in muscle oxidative stress. Novelty: • Our results show for the first time that immunoproteasome and total proteasome function is significantly reduced within obese muscle. • Visceral fat mass is a significant predictor of diminished proteasome activity in skeletal muscle. • Proteasome function is inversely correlated with an intramuscular accumulation of oxidatively damaged proteins.

DOI: https://doi.org/10.1139/apnm-2021-0764
Adv Exp Med Biol. 2021 ;1349 225-247

Physiological and Pathological Relevance of Selective and Nonselective Ca2+ Channels in Skeletal and Cardiac Muscle.

Jaime Balderas-Villalobos, Tyler W E Steele, Jose M Eltit.

  Contraction of the striated muscle is fundamental for human existence. The action of voluntary skeletal muscle enables activities such as breathing, establishing body posture, and diverse body movements. Additionally, highly precise motion empowers communication, artistic expression, and other activities that define everyday human life. The involuntary contraction of striated muscle is the core function of the heart and is essential for blood flow. Several ion channels are important in the transduction of action potentials to cytosolic Ca2+ signals that enable muscle contraction; however, other ion channels are involved in the progression of muscle pathologies that can impair normal life or threaten it. This chapter describes types of selective and nonselective Ca2+ permeable ion channels expressed in the striated muscle, their participation in different aspects of muscle excitation and contraction, and their relevance to the progression of some pathological states.

Keywords:  Calcium; Dystrophy; Hypertrophy; Malignant hyperthermia; Orai1; Resting Ca2+ entry; Ryanodine receptor; STIM1; Store-operated Ca2+ entry; TRPC; Voltage-gated calcium channel

DOI:  https://doi.org/10.1007/978-981-16-4254-8_11
Front Physiol. 2021 ;12 786895

Exercise Capacity Is Improved by Levosimendan in Heart Failure and Sarcopenia via Alleviation of Apoptosis of Skeletal Muscle.

Di Wang, Ming Song, Long-Fei Shen, Lu Han, Ping Zhu, Xu Jia, Guo-Kai Shang, Yuan Cao, Wei Zhang, Ming Zhong, Zhi-Hao Wang.

   Background: Patients suffering from chronic heart failure (CHF) show an increased prevalence of sarcopenia. Levosimendan is an effective drug for the treatment of heart failure, but its effect on sarcopenia is still unclear. We aimed to explore whether levosimendan could enhance skeletal muscle contractibility, improve skeletal muscle atrophy, and thus improve exercise tolerance of individuals with heart failure.
Methods: C57BL6/J mice were used to establish the heart failure with sarcopenia model and injected of levosimendan. Mice were separated into control group, sham operation group, HF group, HF + solvent group, HF + levosimendan group, HF + sarcopenia group, HF + sarcopenia + solvent group, HF + sarcopenia + levosimendan group (n = 5-12). After the treatment, exercise capacity and cardiac function were evaluated. Muscle morphology, inflammation level and apoptosis levels were detected, in which mitochondrial function and oxidative stress level were also assessed.
Result: Levosimendan could increase forelimb grip strength/body weight, hanging impulse, maximum running distance and time in mice with HF and sarcopenia (P < 0.0001 for all), and these improvements were independent of EF (P = 0.0019 for hanging impulse, P < 0.001 for forelimb grip strength/body weight and maximum running distance). Levosimendan directly increased the CSA of gastrocnemius in mice with HF and sarcopenia (P < 0.0001). After levosimendan injection, the proportion of slow muscle fibers increased (P < 0.0001), but this improvement of muscle fiber typing might be attributed to improved cardiac function (P > 0.05). Levosimendan also maintained mitochondrial membrane potential, decreased cleaved caspase-3 (P = 0.034), cleaved caspase-9 (P < 0.0001), Bax expression (P < 0.0001), and increased Bcl2 expression (P = 0.0036). This effect is independent of improved cardiac function (P = 0.028 for bax, P < 0.001 for cleaved caspase-9 and Bcl2). IL-6, TNF-α expression (P < 0.0001 for both) decreased, and SOD activity (P = 0.0038), GSH/GSSG ratio (P = 0.002) significantly increased in skeletal muscle after injection of levosimendan. The improvement in oxidative stress level was attributed to improved cardiac function (P > 0.05).
Conclusion: Levosimendan reduce the loss of skeletal muscle mitochondrial membrane potential, decrease the apoptosis, alleviate the inflammation and oxidative stress, and ultimately improve the exercise capacity of mice with heart failure and sarcopenia. Therefore, levosimendan may be a potential drug for the treatment of heart failure with sarcopenia.

Keywords:  apoptosis; heart failure; levosimendan; mitochondrial function; sarcopenia

DOI:  https://doi.org/10.3389/fphys.2021.786895
J Appl Physiol (1985). 2022 Feb 10.

Acute exercise rapidly activates hepatic mitophagic flux.

Colin S McCoin, Edziu Franczak, Fengyan Deng, Dong Pei, Wen-Xing Ding, John P Thyfault.

  Exercise is critical for improving metabolic health and putatively maintains or enhances mitochondrial quality control in metabolic tissues. While previous work has shown exercise elicits hepatic mitochondrial biogenesis, it is unknown if acute exercise activates hepatic mitophagy, the selective degradation of damaged or low-functioning mitochondria. We tested if an acute bout of treadmill running increased hepatic mitophagic flux both immediately after and 2 hours post-exercise in 15-24-week-old C57BL/6J female mice. Acute exercise did not significantly increase markers of autophagic flux, however, mitophagic flux was activated 2 hours post-treadmill running as measured by accumulation of both LC3-II and p62 in isolated mitochondria in the presence of leupeptin, an inhibitor of autophagosome degradation. Further, mitochondrial associated ubiquitin, which recruits the autophagy receptor protein p62, was also significantly increased at 2 hours. Further examination via western blot and proteomics analysis revealed acute exercise elicits a time-dependent, dynamic activation of mitophagy pathways. Moreover, the results suggest that exercise induced hepatic mitophagy is likely mediated by both poly-ubiquitination and receptor mediated signaling pathways. Overall, we provide evidence that acute exercise activates hepatic mitophagic flux while also revealing specific receptor-mediated proteins by which exercise maintains mitochondrial quality control in the liver.

Keywords:  Exercise; Liver; Mitochondria; Mitophagic Flux; Mitophagy

DOI:  https://doi.org/10.1152/japplphysiol.00704.2021
J Cachexia Sarcopenia Muscle. 2022 Feb 07.

RhoA/ROCK2 signalling is enhanced by PDGF-AA in fibro-adipogenic progenitor cells: implications for Duchenne muscular dystrophy.

Esther Fernández-Simón, Xavier Suárez-Calvet, Ana Carrasco-Rozas, Patricia Piñol-Jurado, Susana López-Fernández, Gemma Pons, Joan Josep Bech Serra, Carolina de la Torre, Noemí de Luna, Eduard Gallardo, Jordi Díaz-Manera.

   BACKGROUND: The lack of dystrophin expression in Duchenne muscular dystrophy (DMD) induces muscle fibre and replacement by fibro-adipose tissue. Although the role of some growth factors in the process of fibrogenesis has been studied, pathways activated by PDGF-AA have not been described so far. Our aim was to study the molecular role of PDGF-AA in the fibrotic process of DMD.
METHODS: Skeletal muscle fibro-adipogenic progenitor cells (FAPs) from three DMD treated with PDGF-AA at 50 ng/mL were analysed by quantitative mass spectrometry-based proteomics. Western-blot, immunofluorescence, and G-LISA were used to confirm the mass spectrometry results. We evaluated the effects of PDGF-AA on the activation of RhoA pathway using two inhibitors, C3-exoenzyme and fasudil. Cell proliferation and migration were determined by BrdU and migration assay. Actin reorganization and collagen synthesis were measured by phalloidin staining and Sircol assay, respectively. In an in vivo proof of concept study, we treated dba/2J-mdx mice with fasudil for 6 weeks. Muscle strength was assessed with the grip strength. Immunofluorescence and flow cytometry analyses were used to study fibrotic and inflammatory markers in muscle tissue.
RESULTS: Mass spectrometry revealed that RhoA pathway proteins were up-regulated in treated compared with non-treated DMD FAPs (n = 3, mean age = 8 ± 1.15 years old). Validation of proteomic data showed that Arhgef2 expression was significantly increased in DMD muscles compared with healthy controls by a 7.7-fold increase (n = 2, mean age = 8 ± 1.14 years old). In vitro studies showed that RhoA/ROCK2 pathway was significantly activated by PDGF-AA (n = 3, 1.88-fold increase, P < 0.01) and both C3-exoenzyme and fasudil blocked that activation (n = 3, P < 0.05 and P < 0.001, respectively). The activation of RhoA pathway by PDGF-AA promoted a significant increase in proliferation and migration of FAPs (n = 3, P < 0.001), while C3-exoenzyme and fasudil inhibited FAPs proliferation at 72 h and migration at 48 and 72 h (n = 3, P < 0.001). In vivo studies showed that fasudil improved muscle function (n = 5 non-treated dba/2J-mdx and n = 6 treated dba/2J-mdx, 1.76-fold increase, P < 0.013), and histological studies demonstrated a 23% reduction of collagen-I expression area (n = 5 non-treated dba/2J-mdx and n = 6 treated dba/2J-mdx, P < 0.01).
CONCLUSIONS: Our results suggest that PDGF-AA promotes the activation of RhoA pathway in FAPs from DMD patients. This pathway could be involved in FAPs activation promoting its proliferation, migration, and actin reorganization, which represents the beginning of the fibrotic process. The inhibition of RhoA pathway could be considered as a potential therapeutic target for muscle fibrosis in patients with muscular dystrophies.

Keywords:  Duchenne muscular dystrophy; Fibro-adipogenic precursor cells; Fibrosis; Muscular dystrophies; Platelet-derived growth factor

DOI:  https://doi.org/10.1002/jcsm.12923
Annu Rev Physiol. 2022 Feb 10. 84 209-227

AMPK and the Adaptation to Exercise.

Hannah R Spaulding, Zhen Yan.

  Noncommunicable diseases are chronic diseases that contribute to death worldwide, but these diseases can be prevented and mitigated with regular exercise. Exercise activates signaling molecules and the transcriptional network to promote physiological adaptations, such as fiber type transformation, angiogenesis, and mitochondrial biogenesis. AMP-activated protein kinase (AMPK) is a master regulator that senses the energy state, promotes metabolism for glucose and fatty acid utilization, and mediates beneficial cellular adaptations in many vital tissues and organs. This review focuses on the current, integrative understanding of the role of exercise-induced activation of AMPK in the regulation of system metabolism and promotion of health benefits.

Keywords:  AMPK; adaptive responses; exercise; fatty acid oxidation; glucose uptake; metabolism

DOI:  https://doi.org/10.1146/annurev-physiol-060721-095517
Math Biosci Eng. 2022 Jan;19(2): 2095-2119

Exploring the role of eRNA in regulating gene expression.

Heli Tan, Tuoqi Liu, Tianshou Zhou.

  eRNAs as the products of enhancers can regulate gene expression via various possible ways, but which regulation way is more reasonable is debatable in biology, and in particular, how eRNAs impact gene expression remains unclear. Here we introduce a mechanistic model of gene expression to address these issues. This model considers three possible regulation ways of eRNA: Type-I by which eRNA regulates transcriptional activity by facilitating the formation of enhancer-promoter (E-P) loop, Type-II by which eRNA directly promotes the mRNA production rate, and mixed regulation (i.e., the combination of Type-I and Type-II). We show that with the increase of the E-P loop length, mRNA distribution can transition from unimodality to bimodality or vice versa in all the three regulation cases. However, in contrast to the other two regulations, Type-II regulation can lead to the highest mean mRNA level and the lowest mRNA noise, independent of the E-P loop length. These results would not only reveal the essential mechanism of how eRNA regulates gene expression, but also imply a new mechanism for phenotypic switching, namely the E-P loop can induce phenotypic switching.

Keywords:   E-P looping ; chemical master equation ; eRNA ; enhancer ; gene expression model

DOI:  https://doi.org/10.3934/mbe.2022098
Eur J Appl Physiol. 2022 Feb 11.

Lean mass sparing in resistance-trained athletes during caloric restriction: the role of resistance training volume.

C Roth, B J Schoenfeld, M Behringer.

  Many sports employ caloric restriction (CR) to reduce athletes' body mass. During these phases, resistance training (RT) volume is often reduced to accommodate recovery demands. Since RT volume is a well-known anabolic stimulus, this review investigates whether a higher training volume helps to spare lean mass during CR. A total of 15 studies met inclusion criteria. The extracted data allowed calculation of total tonnage lifted (repetitions × sets × intensity load) or weekly sets per muscle group for only 4 of the 15 studies, with RT volume being highly dependent on the examined muscle group as well as weekly training frequency per muscle group. Studies involving high RT volume programs (≥ 10 weekly sets per muscle group) revealed low-to-no (mostly female) lean mass loss. Additionally, studies increasing RT volume during CR over time appeared to demonstrate no-to-low lean mass loss when compared to studies reducing RT volume. Since data regarding RT variables applied were incomplete in most of the included studies, evidence is insufficient to conclude that a higher RT volume is better suited to spare lean mass during CR, although data seem to favor higher volumes in female athletes during CR. Moreover, the data appear to suggest that increasing RT volume during CR over time might be more effective in ameliorating CR-induced atrophy in both male and female resistance-trained athletes when compared to studies reducing RT volume. The effects of CR on lean mass sparing seem to be mediated by training experience, pre-diet volume, and energy deficit, with, on average, women tending to spare more lean mass than men. Potential explanatory mechanisms for enhanced lean mass sparing include a preserved endocrine milieu as well as heightened anabolic signaling.

Keywords:  Anabolism; Intracellular pathways; Protein degradation; Protein synthesis; Weight loss; Weight training

DOI:  https://doi.org/10.1007/s00421-022-04896-5
J Biol Chem. 2022 Feb 03. pii: S0021-9258(22)00119-3. [Epub ahead of print] 101679

FNDC5/irisin facilitates muscle-adipose-bone connectivity through ubiquitination-dependent activation of runt-related transcriptional factors RUNX1/2.

Xinyu He, Yue Hua, Qian Li, Wei Zhu, Yu Pan, Yilin Yang, Xinyang Li, Mengxiao Wu, Jiyong Wang, Xiaoqing Gan.

  In the past decade, the cleavage protein irisin derived from fibronectin type III domain-containing protein 5 (FNDC5) in exercise-stimulated skeletal muscle has increasingly become a biomarker associated with metabolic syndrome and osteoporosis in humans. However, it is unclear how this protein facilitates muscle-adipose-bone connectivity in metabolic and skeletal homeostasis. In this study, we unexpectedly observed that the FNDC5 gene can be markedly activated during the differentiation of brown adipocytes but not white adipocytes, and that FNDC5 is specifically expressed in mouse brown adipose tissues (BAT). But unlike it in the skeletal muscles, the expression of FNDC5/irisin in BAT is promoted by cold exposure rather than exercise in mice. Analysis of promoter activity and chromatin immunoprecipitation further showed that that peroxisome proliferator-activated receptor γ (PPARγ) coactivator-1α and thyroid hormone receptors cooperate on the FNDC5 gene promoter to induce its transcription. We found that FNDC5/irisin stimulates the runt-related transcriptional factors RUNX1/2 via a focal adhesion kinase (FAK)-dependent pathway in both bone and subcutaneous white adipose tissues. Mechanistically, we showed that FAK is stimulated by FNDC5/irisin, and then facilitates E3 ubiquitin-protein ligase WW domain-containing protein 2 (WWP2) to ubiquitinate and subsequently activate RUNX1/2, culminating in the activation of osteoblast- or thermogenesis-related genes. Interestingly, the PR domain containing protein 16 (PRDM16) that is crucial for subcutaneous white adipose "browning" and skeletal development was found to form a complex with RUNX1/2 in a WWP2-dependent manner. These findings elucidate a signaling mechanism by which FNDC5/irisin supports the muscle-adipose-bone connectivity, especially BAT-bone connectivity.

Keywords:  WW domain-containing protein 2 (WWP2); brown adipose tissue (BAT); fibronectin type III domain-containing protein 5 (FNDC5)/irisin; focal adhesion kinase (FAK); the runt-related transcriptional factor 1/2 (RUNX1/2); white adipose tissue (WAT)

DOI:  https://doi.org/10.1016/j.jbc.2022.101679
Sci Rep. 2022 Feb 10. 12(1): 2306

Lamina-associated polypeptide 2α is required for intranuclear MRTF-A activity.

Ekaterina Sidorenko, Maria Sokolova, Antti P Pennanen, Salla Kyheröinen, Guido Posern, Roland Foisner, Maria K Vartiainen.

Myocardin-related transcription factor A (MRTF-A), a coactivator of serum response factor (SRF), regulates the expression of many cytoskeletal genes in response to cytoplasmic and nuclear actin dynamics. Here we describe a novel mechanism to regulate MRTF-A activity within the nucleus by showing that lamina-associated polypeptide 2α (Lap2α), the nucleoplasmic isoform of Lap2, is a direct binding partner of MRTF-A, and required for the efficient expression of MRTF-A/SRF target genes. Mechanistically, Lap2α is not required for MRTF-A nuclear localization, unlike most other MRTF-A regulators, but is required for efficient recruitment of MRTF-A to its target genes. This regulatory step takes place prior to MRTF-A chromatin binding, because Lap2α neither interacts with, nor specifically influences active histone marks on MRTF-A/SRF target genes. Phenotypically, Lap2α is required for serum-induced cell migration, and deregulated MRTF-A activity may also contribute to muscle and proliferation phenotypes associated with loss of Lap2α. Our studies therefore add another regulatory layer to the control of MRTF-A-SRF-mediated gene expression, and broaden the role of Lap2α in transcriptional regulation.

DOI: https://doi.org/10.1038/s41598-022-06135-5
Front Mol Biosci. 2021 ;8 812681

Swimming Attenuates Muscle Wasting and Mediates Multiple Signaling Pathways and Metabolites in CT-26 Bearing Mice.

Jiapeng Li, Qiurong Xie, Liya Liu, Ying Cheng, Yuying Han, Xiaoping Chen, Jia Lin, Zuanfang Li, Huixin Liu, Xiuli Zhang, Haichun Chen, Jun Peng, Aling Shen.

  Objectives: To investigate the effects of swimming on cancer induced muscle wasting and explore its underlying mechanism in CT-26 bearing mice. Methods: BALB/c mice (n = 16) injected with CT-26 cells were divided into two groups, including Tumor group (n = 8) and Swimming group (n = 8). Another 8 un-injected mice were set as Control group. Mice in Swimming group were subjected to physical training for swimming twice per day for 30 min intervals and 6 days per week for a total of 4 weeks. The tumor volume was monitored every 3 days and tumor weight was measured at the end of experiment. The changes of muscle function, pathological and cell apoptosis of quadriceps muscles were further assessed, and its underlying mechanisms were further explored using multiple biological technologies. Results: Swimming obviously alleviated tumor volume and weight in CT-26 bearing mice. Moreover, swimming attenuated the decrease of muscle tension, autonomic activities, and increase of muscle atrophy, pathological ultrastructure, as well as cell apoptosis of quadriceps muscles in CT-26 bearing mice. Furthermore, swimming significantly down-regulated the protein expression of NF-κB, p-NF-κB, TNF-α, IL-1β, IL-6 and Bax, while up-regulated the expression of Bcl-2. Further differential expressed metabolites (DEMs) analysis identified a total of 76 (in anion mode) and 330 (in cationic mode) DEMs in quadriceps muscles of CT-26 bearing mice after swimming, including taurochenodeoxycholic acid, taurocholic acid, ascorbic acid and eicosapentaenoic acid. Conclusion: Swimming attenuates tumor growth and muscle wasting, and by suppressing the activation of NF-κB signaling pathway mediated inflammation, reducing the level of Bax medicated cell apoptosis, as well as modulating multiple metabolites might be the importantly underlying mechanisms.

Keywords:  NF-κB; colorectal cancer; metabolite; muscle wasting; swimming

DOI:  https://doi.org/10.3389/fmolb.2021.812681
Neuron. 2022 Feb 04. pii: S0896-6273(22)00049-6. [Epub ahead of print]

Conformational transitions and ligand-binding to a muscle-type nicotinic acetylcholine receptor.

Eleftherios Zarkadas, Eva Pebay-Peyroula, Mackenzie John Thompson, Guy Schoehn, Tomasz Uchański, Jan Steyaert, Christophe Chipot, Francois Dehez, John Edward Baenziger, Hugues Nury.

  Fast synaptic communication requires receptors that respond to the presence of neurotransmitter by opening an ion channel across the post-synaptic membrane. The muscle-type nicotinic acetylcholine receptor from the electric fish, Torpedo, is the prototypic ligand-gated ion channel, yet the structural changes underlying channel activation remain undefined. Here we use cryo-EM to solve apo and agonist-bound structures of the Torpedo nicotinic receptor embedded in a lipid nanodisc. Using both a direct biochemical assay to define the conformational landscape and molecular dynamics simulations to assay flux through the pore, we correlate structures with functional states and elucidate the motions that lead to pore activation of a heteromeric nicotinic receptor. We highlight an underappreciated role for the complementary subunit in channel gating, establish the structural basis for the differential agonist affinities of α/δ versus α /γ sites, and explain why nicotine is less potent at muscle nicotinic receptors compared to neuronal ones.

Keywords:  activation mechanism; agonist binding; cryo-electron miscroscopy; lipid binding; molecular dynamics simulations; nicotine potency; nicotinic acetylcholine receptor; non-equivalent agonist sites; pentameric ligand-gated ion channels; structure and function

DOI:  https://doi.org/10.1016/j.neuron.2022.01.013
Ann Neurol. 2022 Feb 11.

COVID-19-associated Critical Illness Myopathy with Direct Viral Effects.

Dubravka Dodig, Mark A Tarnopolsky, Marta Margeta, Katerina Gordon, Marvin J Fritzler, Jian-Qiang Lu.

COVID-19 (SARS-CoV-2 infection) can lead to intensive care unit (ICU) admission and critical illness myopathy (CIM). We examined three ICU patients with COVID-19, who required mechanical ventilation for pneumonia and developed CIM. Pathological examination of skeletal muscle biopsies revealed myopathic changes consistent with CIM, variable inflammation with autophagic vacuoles, SARS-CoV immunostaining+ fibers/granules, and electron microscopy findings of mitochondrial abnormalities and coronavirus-like particles. While mitochondrial dysfunction with compromised energy production is a critical pathogenic mechanism of non-COVID-19-associated CIM, in our series of COVID-19-associated CIM, myopathic changes including prominent mitochondrial damage suggest a similar mechanism and association with direct SARS-CoV-2 muscle infection. This article is protected by copyright. All rights reserved.

DOI: https://doi.org/10.1002/ana.26318
FASEB J. 2022 Mar;36(3): e22177

Exercise improves angiogenic function of circulating exosomes in type 2 diabetes: Role of exosomal SOD3.

Kareem Abdelsaid, Varadarajan Sudhahar, Ryan A Harris, Archita Das, Seock-Won Youn, Yutao Liu, Maggie McMenamin, Yali Hou, David Fulton, Mark W Hamrick, Yaoliang Tang, Tohru Fukai, Masuko Ushio-Fukai.

  Exosomes, key mediators of cell-cell communication, derived from type 2 diabetes mellitus (T2DM) exhibit detrimental effects. Exercise improves endothelial function in part via the secretion of exosomes into circulation. Extracellular superoxide dismutase (SOD3) is a major secretory copper (Cu) antioxidant enzyme that catalyzes the dismutation of O2 •- to H2 O2 whose activity requires the Cu transporter ATP7A. However, the role of SOD3 in exercise-induced angiogenic effects of circulating plasma exosomes on endothelial cells (ECs) in T2DM remains unknown. Here, we show that both SOD3 and ATP7A proteins were present in plasma exosomes in mice, which was significantly increased after two weeks of volunteer wheel exercise. A single bout of exercise in humans also showed a significant increase in SOD3 and ATP7A protein expression in plasma exosomes. Plasma exosomes from T2DM mice significantly reduced angiogenic responses in human ECs or mouse skin wound healing models, which was associated with a decrease in ATP7A, but not SOD3 expression in exosomes. Exercise training in T2DM mice restored the angiogenic effects of T2DM exosomes in ECs by increasing ATP7A in exosomes, which was not observed in exercised T2DM/SOD3-/- mice. Furthermore, exosomes overexpressing SOD3 significantly enhanced angiogenesis in ECs by increasing local H2 O2 levels in a heparin-binding domain-dependent manner as well as restored defective wound healing and angiogenesis in T2DM or SOD3-/- mice. In conclusion, exercise improves the angiogenic potential of circulating exosomes in T2DM in a SOD3-dependent manner. Exosomal SOD3 may provide an exercise mimetic therapy that supports neovascularization and wound repair in cardiometabolic disease.

Keywords:  SOD3; exercise; exosome; type 2 diabetes

DOI:  https://doi.org/10.1096/fj.202101323R