bims-moremu 2021-09-05 papers

bims-moremu

Biomed News

on Molecular regulators of muscle mass

Issue of 2021–09–05
thirty-six papers selected by
Anna Vainshtein, Craft Science Inc.

Age-Related Increase in Lactate Dehydrogenase Activity in Skeletal Muscle Reduces Lifespan in Drosophila.
Functional significance of gain-of-function H19 lncRNA in skeletal muscle differentiation and anti-obesity effects.
What's all the hype with fiber type? selective single fiber adaptations with lifelong endurance exercise.
The Satellite Cell at 60: The Foundation Years.
The Role of the Skeletal Muscle Secretome in Mediating Endurance and Resistance Training Adaptations.
A single bout of cycling exercise induces nucleosome repositioning in skeletal muscle of lean and obese individuals.
Effects of functional electrical stimulation on muscle health after spinal cord injury.
Skeletal muscle energy metabolism in obesity.
Maslinic acid activates mTORC1 and human TGR5 and induces skeletal muscle hypertrophy.
Deletion of phosphatidylserine flippase β-subunit Tmem30a in satellite cells leads to delayed skeletal muscle regeneration.
Sarcopenia; functional concerns, molecular mechanisms involved, and seafood as a nutritional intervention - review article.
A muscle-centric view of time-restricted feeding for older adults.
Genetic variability affects the response of skeletal muscle to disuse.
mTORC2 is an important target for simvastatin-associated toxicity in C2C12 cells and mouse skeletal muscle - roles of Rap1 geranylgeranylation and mitochondrial dysfunction.
Three-dimensional in vitro models of neuromuscular tissue.
A carbonized wormwood modified photothermal microneedle patch for the repair of damaged skeletal muscles.
Early Changes in Skeletal Muscle of Young C22 Mice, A Model of Charcot-Marie-Tooth 1A.
Myofiber / pro-inflammatory macrophage interplay controls muscle damage in mdx mice.
Simvastatin does not alleviate muscle pathology in a mouse model of Duchenne muscular dystrophy.
DHA but not EPA induces the trans-differentiation of C2C12 cells into white-like adipocytes phenotype.
The effects of pulmonary hypertension on skeletal muscle oxygen pressures in contracting rat spinotrapezius muscle.
High-resolution structure of the membrane-embedded skeletal muscle ryanodine receptor.
Mapping Co-regulatory Interactions among Ligand Binding sites in RyR1.
Acetyl-CoA carboxylase 2 inhibition reduces skeletal muscle bioactive lipid content and attenuates progression of type 2 diabetes in Zucker diabetic fatty rats.
Skeletal muscle is an early site of Zika virus replication and injury impairing myogenesis.
Emerging mechanisms for exercise effects on muscle wasting and anabolic resistance.
Dissection of Single Skeletal Muscle Fibers for Immunofluorescent and Morphometric Analyses of Whole-Mount Neuromuscular Junctions.
Can increasing physical activity prevent aging-related loss of skeletal muscle?
Minimally invasive co-injection of modular micro-muscular and micro-vascular tissues improves in situ skeletal muscle regeneration.
Muscle-directed AAV gene therapy rescues the maple syrup urine disease phenotype in a mouse model.
Diet and exercise reduce pre-existing NASH and fibrosis and have additional beneficial effects on the vasculature, adipose tissue and skeletal muscle via organ-crosstalk.
Physiological complexity: Influence of ageing, disease and neuromuscular fatigue on muscle force and torque fluctuations.
High levels of TFAM repress mammalian mitochondrial DNA transcription in vivo.
RNA-bound PGC-1α controls gene expression in liquid-like nuclear condensates.
Muscle-specific economy of force generation and efficiency of work production during human running.
RGS12 is required for the maintenance of mitochondrial function during skeletal development.

J Gerontol A Biol Sci Med Sci. 2021 Sep 03. pii: glab260. [Epub ahead of print]

Age-Related Increase in Lactate Dehydrogenase Activity in Skeletal Muscle Reduces Lifespan in Drosophila.

Liam C Hunt, Fabio Demontis.

  Metabolic adaptations occur with aging but the significance and causal roles of such changes are only partially known. In Drosophila, we find that skeletal muscle aging is paradoxically characterized by increased readouts of glycolysis (lactate, NADH/NAD+) but reduced expression of most glycolytic enzymes. This conundrum is explained by lactate dehydrogenase (LDH), an enzyme necessary for anaerobic glycolysis and whose expression increases with aging. Experimental Ldh overexpression in skeletal muscle of young flies increases glycolysis and shortens lifespan, suggesting that age-related increases in muscle LDH contribute to mortality. Similar results are also found with overexpression of other glycolytic enzymes (Pfrx/PFKFB, Pgi/GPI). Conversely, hypomorphic mutations in Ldh extend lifespan whereas reduction in PFK, Pglym78/PGAM, Pgi/GPI, and Ald/ALDO levels shorten lifespan to various degrees, indicating that glycolysis needs to be tightly controlled for optimal aging. Altogether, these findings indicate a role for muscle LDH and glycolysis in aging.

Keywords:  Drosophila; LDH; aging; glycolysis; lifespan; skeletal muscle

DOI:  https://doi.org/10.1093/gerona/glab260
Genome Med. 2021 Aug 28. 13(1): 137

Functional significance of gain-of-function H19 lncRNA in skeletal muscle differentiation and anti-obesity effects.

Yajuan Li, Yaohua Zhang, Qingsong Hu, Sergey D Egranov, Zhen Xing, Zhao Zhang, Ke Liang, Youqiong Ye, Yinghong Pan, Sujash S Chatterjee, Brandon Mistretta, Tina K Nguyen, David H Hawke, Preethi H Gunaratne, Mien-Chie Hung, Leng Han, Liuqing Yang, Chunru Lin.

   BACKGROUND: Exercise training is well established as the most effective way to enhance muscle performance and muscle building. The composition of skeletal muscle fiber type affects systemic energy expenditures, and perturbations in metabolic homeostasis contribute to the onset of obesity and other metabolic dysfunctions. Long noncoding RNAs (lncRNAs) have been demonstrated to play critical roles in diverse cellular processes and diseases, including human cancers; however, the functional importance of lncRNAs in muscle performance, energy balance, and obesity remains elusive. We previously reported that the lncRNA H19 regulates the poly-ubiquitination and protein stability of dystrophin (DMD) in muscular dystrophy.
METHODS: Here, we identified mouse/human H19-interacting proteins using mouse/human skeletal muscle tissues and liquid chromatography-mass spectrometry (LC-MS). Human induced pluripotent stem-derived skeletal muscle cells (iPSC-SkMC) from a healthy donor and Becker Muscular Dystrophy (BMD) patients were utilized to study DMD post-translational modifications and associated proteins. We identified a gain-of-function (GOF) mutant of H19 and characterized the effects on myoblast differentiation and fusion to myotubes using iPSCs. We then conjugated H19 RNA gain-of-function oligonucleotides (Rgof) with the skeletal muscle enrichment peptide agrin (referred to as AGR-H19-Rgof) and evaluated AGR-H19-Rgof's effects on skeletal muscle performance using wild-type (WT) C57BL/6 J mice and its anti-obesity effects using high-fat diet (HFD)- and leptin deficiency-induced obese mouse models.
RESULTS: We demonstrated that both human and mouse H19 associated with DMD and that the H19 GOF exhibited enhanced interaction with DMD compared to WT H19. DMD was found to associate with serine/threonine-protein kinase MRCK alpha (MRCKα) and α-synuclein (SNCA) in iPSC-SkMC derived from BMD patients. Inhibition of MRCKα and SNCA-mediated phosphorylation of DMD antagonized the interaction between H19 and DMD. These signaling events led to improved skeletal muscle cell differentiation and myotube fusion. The administration of AGR-H19-Rgof improved the muscle mass, muscle performance, and base metabolic rate of WT mice. Furthermore, mice treated with AGR-H19-Rgof exhibited resistance to HFD- or leptin deficiency-induced obesity.
CONCLUSIONS: Our study suggested the functional importance of the H19 GOF mutant in enhancing muscle performance and anti-obesity effects.

Keywords:  Dystrophin; H19; Long noncoding RNA; Obesity; RNA therapy; Skeletal muscle

DOI:  https://doi.org/10.1186/s13073-021-00937-4
J Physiol. 2021 Aug 31.

What's all the hype with fiber type? selective single fiber adaptations with lifelong endurance exercise.

Meral N Culver, Sean P Langan, Zach J Hutchison.



Keywords:  aging; exercise; performance; single fiber; skeletal muscle

DOI:  https://doi.org/10.1113/JP281986
J Neuromuscul Dis. 2021 Aug 27.

The Satellite Cell at 60: The Foundation Years.

Elise N Engquist, Peter S Zammit.

  The resident stem cell for skeletal muscle is the satellite cell. On the 50th anniversary of its discovery in 1961, we described the history of skeletal muscle research and the seminal findings made during the first 20 years in the life of the satellite cell (Scharner and Zammit 2011, doi: 10.1186/2044-5040-1-28). These studies established the satellite cell as the source of myoblasts for growth and regeneration of skeletal muscle. Now on the 60th anniversary, we highlight breakthroughs in the second phase of satellite cell research from 1980 to 2000. These include technical innovations such as isolation of primary satellite cells and viable muscle fibres complete with satellite cells in their niche, and generation of many useful reagents including genetically modified organisms and antibodies still in use today. New methodologies were combined with description of endogenous satellite cells markers, notably Pax7. Discovery of the muscle regulatory factors Myf5, MyoD, Myogenin, and MRF4 in the late 1980s revolutionized understanding of the control of both developmental and regerenative myogenesis. Emergence of genetic lineage markers facilitated identification of satellite cells in situ, but also empowered transplantation studies to examine satellite cell function. Finally, satellite cell heterogeneity and the non-satellite cell support of muscle regeneration were described. These major advances in methodology and in understanding satellite cell biology provided the foundations for the dramatic escalation of work on muscle stem cells in the 21st century.

Keywords:  Myf5; MyoD; PAX7; Satellite cell; muscle regeneration; muscle regulatory factors; myofibre; skeletal muscle

DOI:  https://doi.org/10.3233/JND-210705
Front Physiol. 2021 ;12 709807

The Role of the Skeletal Muscle Secretome in Mediating Endurance and Resistance Training Adaptations.

Aurel B Leuchtmann, Volkan Adak, Sedat Dilbaz, Christoph Handschin.

  Exercise, in the form of endurance or resistance training, leads to specific molecular and cellular adaptions not only in skeletal muscles, but also in many other organs such as the brain, liver, fat or bone. In addition to direct effects of exercise on these organs, the production and release of a plethora of different signaling molecules from skeletal muscle are a centerpiece of systemic plasticity. Most studies have so far focused on the regulation and function of such myokines in acute exercise bouts. In contrast, the secretome of long-term training adaptation remains less well understood, and the contribution of non-myokine factors, including metabolites, enzymes, microRNAs or mitochondrial DNA transported in extracellular vesicles or by other means, is underappreciated. In this review, we therefore provide an overview on the current knowledge of endurance and resistance exercise-induced factors of the skeletal muscle secretome that mediate muscular and systemic adaptations to long-term training. Targeting these factors and leveraging their functions could not only have broad implications for athletic performance, but also for the prevention and therapy in diseased and elderly populations.

Keywords:  PGC-1alpha; endurance training; exercise; myokines; resistance training; secretome; skeletal muscle

DOI:  https://doi.org/10.3389/fphys.2021.709807
Diabetes Obes Metab. 2021 Sep 02.

A single bout of cycling exercise induces nucleosome repositioning in skeletal muscle of lean and obese individuals.

Prasad P Devarshi, Andrea S Pereyra, Jessica M Ellis, Tara M Henagan.

Acute aerobic exercise improves, whereas being overweight impairs, skeletal muscle mitochondrial function in correlation with reduced insulin sensitivity. Here, the molecular and metabolic effects of a single exercise bout in the skeletal muscle was compared in men, aged 19-30 years, that were either lean (BMI<25, 18.5-24.1 kg/m2 , n=15) or overweight/obese (Ov/Ob; BMI≥25, 25.5-36.9 kg/m2 , n=15). Four hours after a high-carbohydrate breakfast (7 kcal/kg; 60% carbohydrate, 25% fat, 15% protein), participants performed cycling exercise (50% VO2 max, expending ~650 kcal). Muscle biopsies and peripheral blood samples were collected 30 minutes before the meal and immediately after exercise. Muscle long-chain acylcarnitines were increased in Ov/Ob compared to lean, with or without exercise. A single exercise bout increased mRNA abundance of genes related to mitochondria and insulin signaling in both lean and Ov/Ob. Nucleosome mapping by micrococcal nuclease digestion with deep sequencing (MNase-seq) revealed that exercise repositioned the -1N nucleosome away from the transcription start site of the PGC1a promoter and of other mitochondrial genes, but did not affect genes related to insulin signaling, in both lean and Ov/Ob. In conclusion, these data suggest that a single exercise bout induced epigenetic alterations in skeletal muscle in a BMI-independent manner. This article is protected by copyright. All rights reserved.

DOI: https://doi.org/10.1111/dom.14541
Curr Opin Pharmacol. 2021 Aug 28. pii: S1471-4892(21)00124-7. [Epub ahead of print]60 226-231

Effects of functional electrical stimulation on muscle health after spinal cord injury.

Kelly D Atkins, C Scott Bickel.

  Spinal cord injury is a devastating condition interrupting voluntary movement and motor control. In response to unloading, skeletal muscle undergoes numerous adaptations, including rapid and profound atrophy, intramuscular fat accumulation, impaired muscular glucose metabolism and decreased force generation and muscle performance. Functional electrical stimulation (FES) involves electrically stimulating affected muscles to contract in a coordinated manner to create a functional movement or task. Effects of FES-cycling, rowing and resistance training on muscle health are described here. Briefly, FES-cycling and resistance training may slow muscle atrophy or facilitate muscle hypertrophy, and all modalities benefit muscle composition and performance to some extent. These interventions show promise as future rehabilitative tools after spinal cord injury.

Keywords:  Atrophy; Exercise; Functional electrical stimulation; Muscle health; Spinal cord injury

DOI:  https://doi.org/10.1016/j.coph.2021.07.025
Obesity (Silver Spring). 2021 Aug 31.

Skeletal muscle energy metabolism in obesity.

Abel M Mengeste, Arild C Rustan, Jenny Lund.

Comparing energy metabolism in human skeletal muscle and primary skeletal muscle cells in obesity, while focusing on glucose and fatty acid metabolism, shows many common changes. Insulin-mediated glucose uptake in skeletal muscle and primary myotubes is decreased by obesity, whereas differences in basal glucose metabolism are inconsistent among studies. With respect to fatty acid metabolism, there is an increased uptake and storage of fatty acids and a reduced complete lipolysis, suggesting alterations in lipid turnover. In addition, fatty acid oxidation is decreased, probably at the level of complete oxidation, as β -oxidation may be enhanced in obesity, which indicates mitochondrial dysfunction. Metabolic changes in skeletal muscle with obesity promote metabolic inflexibility, ectopic lipid accumulation, and formation of toxic lipid intermediates. Skeletal muscle also acts as an endocrine organ, secreting myokines that participate in interorgan cross talk. This review highlights interventions and some possible targets for treatment through action on skeletal muscle energy metabolism. Effects of exercise in vivo on obesity have been compared with simulation of endurance exercise in vitro on myotubes (electrical pulse stimulation). Possible pharmaceutical targets, including signaling pathways and drug candidates that could modify lipid storage and turnover or increase mitochondrial function or cellular energy expenditure through adaptive thermogenic mechanisms, are discussed.

DOI: https://doi.org/10.1002/oby.23227
Biosci Biotechnol Biochem. 2021 Aug 30. pii: zbab151. [Epub ahead of print]

Maslinic acid activates mTORC1 and human TGR5 and induces skeletal muscle hypertrophy.

Shotaro Murata, Takashi Sasaki, Yuki Yamauchi, Makoto Shimizu, Ryuichiro Sato.

  Maslinic acid, a naturally occurring pentacyclic triterpene in more than 30 plants (including olives), reportedly increases human muscle mass and muscle strength; however, the underlying molecular mechanism remains unknown. C57BL/6J mice were fed a standard diet or supplemented with 0.27% maslinic acid for four weeks, and their skeletal muscle mass was measured. Mice that consumed maslinic acid displayed significant increases in gastrocnemius and soleus muscle mass. Cultured mouse-C2C12 skeletal muscle cells were treated with mammalian target of rapamycin complex 1 (mTORC1) or protein kinase b (Akt) inhibitor, and protein synthesis was quantified. Maslinic acid accelerated protein synthesis via mTORC1 activation independent of Akt. Furthermore, maslinic acid activated human Takeda G protein-coupled receptor 5 (TGR5) more strongly than mouse TGR5, augmenting the expression of several genes related to muscular hypertrophy. Maslinic acid activated mTORC1 and human TGR5, implying its contribution to human muscular hypertrophy through these effects.

Keywords:  TGR5; mammalian target of rapamycin complex 1; maslinic acid; muscle hypertrophy; triterpenoid

DOI:  https://doi.org/10.1093/bbb/zbab151
Zool Res. 2021 Sep 18. pii: 2095-8137(2021)05-0650-10. [Epub ahead of print]42(5): 650-659

Deletion of phosphatidylserine flippase β-subunit Tmem30a in satellite cells leads to delayed skeletal muscle regeneration.

Kuan-Xiang Sun, Xiao-Yan Jiang, Xiao Li, Yu-Jing Su, Ju-Lin Wang, Lin Zhang, Ye-Ming Yang, Xian-Jun Zhu.

  Phosphatidylserine (PS) is distributed asymmetrically in the plasma membrane of eukaryotic cells. Phosphatidylserine flippase (P4-ATPase) transports PS from the outer leaflet of the lipid bilayer to the inner leaflet of the membrane to maintain PS asymmetry. The β subunit TMEM30A is indispensable for transport and proper function of P4-ATPase. Previous studies have shown that the ATP11A and TMEM30A complex is the molecular switch for myotube formation. However, the role of Tmem30a in skeletal muscle regeneration remains elusive. In the current study, Tmem30a was highly expressed in the tibialis anterior (TA) muscles of dystrophin-null ( mdx) mice and BaCl 2-induced muscle injury model mice. We generated a satellite cell (SC)-specific Tmem30a conditional knockout (cKO) mouse model to investigate the role of Tmem30a in skeletal muscle regeneration. The regenerative ability of cKO mice was evaluated by analyzing the number and diameter of regenerated SCs after the TA muscles were injured by BaCl 2-injection. Compared to the control mice, the cKO mice showed decreased Pax7 + and MYH3 + SCs, indicating diminished SC proliferation, and decreased expression of muscular regulatory factors (MYOD and MYOG), suggesting impaired myoblast proliferation in skeletal muscle regeneration. Taken together, these results demonstrate the essential role of Tmem30a in skeletal muscle regeneration.

Keywords:  Atp11a; Knockout mouse model; Satellite cell; Skeletal muscle regeneration; Tmem30a

DOI:  https://doi.org/10.24272/j.issn.2095-8137.2021.195
Crit Rev Food Sci Nutr. 2021 Aug 28. 1-21

Sarcopenia; functional concerns, molecular mechanisms involved, and seafood as a nutritional intervention - review article.

Thilina U Jayawardena, Seo-Young Kim, You-Jin Jeon.

  The fundamental basis for the human function is provided by skeletal muscle. Advancing age causes selective fiber atrophy, motor unit loss, and hybrid fiber formation resulting in hampered mass and strength, thus referred to as sarcopenia. Influence on the loss of independence of aged adults, contribute toward inclined healthcare costs conveys the injurious impact. The current understating of age-related skeletal muscle changes are addressed in this review, and further discusses mechanisms regulating protein turnover, although they do not completely define the process yet. Moreover, the reduced capacity of muscle regeneration due to impairment of satellite cell activation and proliferation with neuronal, immunological, hormonal factors were brought into the light of attention. Nevertheless, complete understating of sarcopenia requires disentangling it from disuse and disease. Nutritional intervention is considered a potentially preventable factor contributing to sarcopenia. Seafood is a crucial player in the fight against hunger and malnutrition, where it consists of macro and micronutrients. Hence, the review shed light on seafood as a nutritional intrusion in the treatment and prevention of sarcopenia. Understanding multiple factors will provide therapeutic targets in the prevention, treatment, and overcoming adverse effects of sarcopenia.

Keywords:  aging; muscle growth; nutrition; sarcopenia; seafood

DOI:  https://doi.org/10.1080/10408398.2021.1969889
Curr Opin Clin Nutr Metab Care. 2021 Sep 01.

A muscle-centric view of time-restricted feeding for older adults.

Matthew J Lees, Nathan Hodson, Daniel R Moore.

PURPOSE OF REVIEW: The practice of time-restricted feeding (TRF) has received fervent interest in recent years as a strategy to mitigate obesity and metabolic disease. We sought to review the implications of TRF for skeletal muscle health and function in aging.
RECENT FINDINGS: TRF has high adherence and can promote body weight loss in older populations. Body weight reductions favor fat mass in the young, however, there is also the potential for undesirable losses in lean mass. There is currently no evidence to support TRF for skeletal muscle function and metabolism in older persons, and only tentative findings in the young. With a narrow eating window of 6-8 h and a prolonged fasting period to minimize daily insulin exposure, TRF may contradict recommended dietary practices for optimizing skeletal muscle anabolism in older people.
SUMMARY: TRF might represent a promising intervention to address obesity and its associated metabolic diseases, however, at present there is insufficient evidence for optimizing skeletal muscle mass or health in older individuals. Further research is needed to: (1) ascertain the impact of TRF on body composition, skeletal muscle anabolism, and autophagy in aging, and; (2) delineate the potentially myoprotective roles of dietary protein and exercise within the framework of TRF in older persons.

DOI: https://doi.org/10.1097/MCO.0000000000000789
J Musculoskelet Neuronal Interact. 2021 Sep 01. 21(3): 387-396

Genetic variability affects the response of skeletal muscle to disuse.

Camilla Reina Maroni, Michael A Friedman, Yue Zhang, Michael J McClure, Stefania Fulle, Charles R Farber, Henry J Donahue.

OBJECTIVE: To examine whether genetic variability plays a role in skeletal muscle response to disuse.
METHODS: We examined skeletal muscle response to disuse in five different strains of mice: CAST/EiJ, NOD/ShiLtJ, NZO/HILtJ, 129S1/SvImJ and A/J. Mice had one limb immobilized by a cast for three weeks.
RESULTS: Response to immobilization was dependent on the strain of mice. Skeletal muscle mass/body weight was decreased by immobilization in all strains except 1291/SvImJ. Immobilization decreased absolute skeletal muscle mass in quadriceps and gastrocnemius in NOD/ShiltJ and NZO/HILtJ mice. Three weeks of immobilization resulted in an increase in quadriceps levels of atrogenes in CAST/EiJ. Immobilization resulted in an increase in quadriceps and gastrocnemius levels of Myh4 in CAST/EiJ. A similar trend was observed for Myh7 in gastrocnemius muscle. Immobilization resulted in a decrease of the p-p70S6K1/total p706SK1 ratio in quadriceps of NOD/ShiLtJ mice and the gastrocnemius of A/J mice. Immobilization did not affect the p-4EBP1/total 4EBP1 ratio in quadriceps of any of the strains examined. However, the p-4EBP1/total 4EBP1 ratio in gastrocnemius was greater in immobilized, relative to control, limbs in CAST/EiJ mice.
CONCLUSION: Genetic variability affects the response of skeletal muscle to disuse.

Keywords: Immobilization; Skeletal Muscle Atrophy
Biochem Pharmacol. 2021 Aug 27. pii: S0006-2952(21)00366-X. [Epub ahead of print] 114750

mTORC2 is an important target for simvastatin-associated toxicity in C2C12 cells and mouse skeletal muscle - roles of Rap1 geranylgeranylation and mitochondrial dysfunction.

Gerda M Sanvee, Leonie Hitzfeld, Jamal Bouitbir, Stephan Krähenbühl.

  Statins decrease the serum LDL-cholesterol concentration and reduce the risk for cardiovascular diseases but can cause myopathy, which may be related to mTORC inhibition. In the current study, we investigated which mTORC is inhibited by simvastatin and by which mechanisms. In C2C12 myoblasts and myotubes and mouse gastrocnemius, simvastatin was cytotoxic and inhibited S6rp and Akt Ser473 phosphorylation, indicating inhibition of mTORC1 and mTORC2, respectively. In contrast to simvastatin, the mTORC1 inhibitor rapamycin did not inhibit mTORC2 activity and was not cytotoxic. Like simvastatin, knock-down of Rictor, an essential component of mTORC2, impaired Akt Ser473 and S6rp phosphorylation and was cytotoxic for C2C12 myoblasts, suggesting that mTORC2 inhibition is an important myotoxic mechanism. The investigation of the mechanism of mTORC2 inhibition showed that simvastatin impaired Ras farnesylation, which was prevented by farnesol but without restoring mTORC2 activity. In comparison, Rap1 knock-down reduced mTORC2 activity and was cytotoxic for C2C12 myoblasts. Simvastatin impaired Rap1 geranylgeranylation and function, which was prevented by geranylgeraniol. In addition, simvastatin and the complex III inhibitor antimycin A caused mitochondrial superoxide accumulation and impaired the activity of mTORC2, which could partially be prevented by the antioxidant MitoTEMPO. In conclusion, mTORC2 inhibition is an important mechanism of simvastatin-induced myotoxicity. Simvastatin inhibits mTORC2 by impairing geranylgeranylation of Rap1 and by inducing mitochondrial dysfunction.

Keywords:  Akt/PKB; Rap1; Rictor; geranylgeranylation; mTORC1; mTORC2; mitochondrial function; simvastatin

DOI:  https://doi.org/10.1016/j.bcp.2021.114750
Neural Regen Res. 2022 Apr;17(4): 759-766

Three-dimensional in vitro models of neuromuscular tissue.

Paolo Raffa, Maria Easler, Anna Urciuolo.

  Skeletal muscle is a dynamic tissue in which homeostasis and function are guaranteed by a very defined three-dimensional organization of myofibers in respect to other non-muscular components, including the extracellular matrix and the nervous network. In particular, communication between myofibers and the nervous system is essential for the overall correct development and function of the skeletal muscle. A wide range of chronic, acute and genetic-based human pathologies that lead to the alteration of muscle function are associated with modified preservation of the fine interaction between motor neurons and myofibers at the neuromuscular junction. Recent advancements in the development of in vitro models for human skeletal muscle have shown that three-dimensionality and integration of multiple cell types are both key parameters required to unveil pathophysiological relevant phenotypes. Here, we describe recent achievement reached in skeletal muscle modeling which used biomaterials for the generation of three-dimensional constructs of myotubes integrated with motor neurons.

Keywords:  3D organization; bioengineering; biomaterials; motor neurons; neuromuscular junction; skeletal muscle models

DOI:  https://doi.org/10.4103/1673-5374.322447
J Mater Chem B. 2021 Sep 03.

A carbonized wormwood modified photothermal microneedle patch for the repair of damaged skeletal muscles.

Chuxi Zhang, Shuang Jia, Jinlong Huang, Haichuan Peng, Jiao Zhang, Lubing Liu, Wei Zhang, Hongbo Xin, Xiaolei Wang.

In this study, we aimed to achieve an efficient repair of damaged skeletal muscles using polyvinyl alcohol (PVA) soluble microneedle patches (MNP) loaded with carbonized wormwood and prostaglandin E2 (inflammatory factors). The introduction of carbonized wormwood imparted the MNP with near-infrared light heating characteristics that improved the efficiency of prostaglandin E2 delivery while also promoting circulation in the damaged muscle area. Our experimental results showed that, compared with the classical moxibustion treatment, the system could more quickly restore muscle strength and the cross-sectional area of muscle bundle fibers in a mouse model of muscular injury. In addition, it could also successfully induce the proliferation and differentiation of muscle stem cells to effectively repair injured muscle tissues. Above all, this light-controlled photothermal MN (microneedle) drug-delivery system avoided the common problems of traditional moxibustion such as large levels of smoke, slow efficacy and risk of scalding. Collectively, we put forward a safe, accurate and efficient approach for skeletal muscle damage treatment using carbonized wormwood.

DOI: https://doi.org/10.1039/d1tb00610j
J Neuromuscul Dis. 2021 Aug 24.

Early Changes in Skeletal Muscle of Young C22 Mice, A Model of Charcot-Marie-Tooth 1A.

Friederike Deres, Stephanie Schwartz, Karin Kappes-Horn, Cornela Kornblum, Jens Reimann.

   BACKGROUND: The C22 mouse is a Charcot-Marie-Tooth 1A transgenic model with minimal axonal loss.
OBJECTIVE: To analyse early skeletal muscle changes resulting from this dysmyelinating neuropathy.
METHODS: Histology of tibialis anterior muscles of C22 mice and wild type litter mate controls for morphometric analysis and (immuno-)histochemistry for known denervation markers and candidate proteins identified by representational difference analysis (RDA) based on mRNA from the same muscles; quantitative PCR and Western blotting for confirmation of RDA findings.
RESULTS: At age 10 days, morphometry was not different between groups, while at 21 days, C22 showed significantly more small diameter fibres, indicating the onset of atrophy at an age when weakness becomes detectable. Neither (immuno-)histochemistry nor RDA detected extrajunctional expression of acetylcholine receptors by age 10 and 21 days, respectively. RDA identified some mRNA up-regulated in C22 muscles, among them at 10 days, prior to detectable weakness or atrophy, integral membrane protein 2a (Itm2a), eukaryotic initiation factor 2, subunit 2 (Eif2s2) and cytoplasmic phosphatidylinositol transfer protein 1 (Pitpnc1). However, qPCR failed to measure significant differences. In contrast, Itm2a and Eif2s2 mRNA were significantly down-regulated comparing 21 versus 10 days of age in both groups, C22 and controls. Western blotting confirmed significant down-regulation of ITM2A protein in C22 only.
CONCLUSION: Denervation-like changes in this model develop slowly with onset of atrophy and weakness at about three weeks of age, before detection of extrajunctional acetylcholine receptors. Altered Itm2a expression seems to being early as an increase, but becomes distinct as a decrease later.

Keywords:  Skeletal muscle; beta subunit; charcot-marie-tooth disease; cytoplasmic 1; demyelination; eukaryotic initiation factor 2; gene expression profiling; integral membrane protein 2A; muscle denervation; phosphatidylinositol transfer protein

DOI:  https://doi.org/10.3233/JND-210681
J Cell Sci. 2021 Sep 02. pii: jcs.258429. [Epub ahead of print]

Myofiber / pro-inflammatory macrophage interplay controls muscle damage in mdx mice.

Marielle Saclier, Sabrina Ben Larbi, Ha My Ly, Eugénie Moulin, Rémi Mounier, Bénédicte Chazaud, Gaëtan Juban.

  Duchenne Muscular Dystrophy is a genetic muscle disease characterized by chronic inflammation and fibrosis, mediated by a pro-fibrotic macrophage population expressing pro-inflammatory markers. Our aim was to characterize cellular events leading to the alteration of macrophage properties, and to modulate macrophage inflammatory status using the gaseous mediator H2S. Using co-culture experiments, we first showed that myofibers derived from mdx mice strongly skewed the polarization of resting macrophages towards a pro-inflammatory phenotype. Treatment of mdx mice with NaHS, an H2S donor, reduced the number of pro-inflammatory macrophages in skeletal muscle, which was associated with a decreased number of nuclei per fiber, together with a reduced myofiber branching and fibrosis. Finally, we established the metabolic sensor AMPK as a critical NaHS target in muscle macrophages. These results identify an interplay between myofibers and macrophages where dystrophic myofibers contribute to the maintenance of a highly inflammatory environment sustaining a pro-inflammatory macrophage status, in turn favoring myofiber damage, myofiber branching and fibrosis establishment. They also identify H2S donors as a potential therapeutic strategy to improve dystrophic muscle phenotype by dampening chronic inflammation.

Keywords:  AMPK; Duchenne muscular dystrophy; Fibrosis; Macrophage; Myofiber branching

DOI:  https://doi.org/10.1242/jcs.258429
Skelet Muscle. 2021 Sep 03. 11(1): 21

Simvastatin does not alleviate muscle pathology in a mouse model of Duchenne muscular dystrophy.

Olga Mucha, Paulina Podkalicka, Katarzyna Kaziród, Emilia Samborowska, Józef Dulak, Agnieszka Łoboda.

   BACKGROUND: Duchenne muscular dystrophy (DMD) is an incurable disease, caused by the mutations in the DMD gene, encoding dystrophin, an actin-binding cytoskeletal protein. Lack of functional dystrophin results in muscle weakness, degeneration, and as an outcome cardiac and respiratory failure. As there is still no cure for affected individuals, the pharmacological compounds with the potential to treat or at least attenuate the symptoms of the disease are under constant evaluation. The pleiotropic agents, 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors, known as statins, have been suggested to exert beneficial effects in the mouse model of DMD. On the other hand, they were also reported to induce skeletal-muscle myopathy. Therefore, we decided to verify the hypothesis that simvastatin may be considered a potential therapeutic agent in DMD.
METHODS: Several methods including functional assessment of muscle function via grip strength measurement, treadmill test, and single-muscle force estimation, enzymatic assays, histological analysis of muscle damage, gene expression evaluation, and immunofluorescence staining were conducted to study simvastatin-related alterations in the mdx mouse model of DMD.
RESULTS: In our study, simvastatin treatment of mdx mice did not result in improved running performance, grip strength, or specific force of the single muscle. Creatine kinase and lactate dehydrogenase activity, markers of muscle injury, were also unaffected by simvastatin delivery in mdx mice. Furthermore, no significant changes in inflammation, fibrosis, and angiogenesis were noted. Despite the decreased percentage of centrally nucleated myofibers in gastrocnemius muscle after simvastatin delivery, no changes were noticed in other regeneration-related parameters. Of note, even an increased rate of necrosis was found in simvastatin-treated mdx mice.
CONCLUSION: In conclusion, our study revealed that simvastatin does not ameliorate DMD pathology.

Keywords:  3-Hydroxy-3-methylglutaryl coenzyme A reductase inhibitors; Angiogenesis; DMD; Duchenne muscular dystrophy; Simvastatin; mdx

DOI:  https://doi.org/10.1186/s13395-021-00276-3
PLoS One. 2021 ;16(9): e0249438

DHA but not EPA induces the trans-differentiation of C2C12 cells into white-like adipocytes phenotype.

Saeed Ghnaimawi, Lisa Rebello, Jamie Baum, Yan Huang.

Muscle derived stem cells (MDSCs) and myoblast play an important role in myotube regeneration when muscle tissue is injured. However, these cells can be induced to differentiate into adipocytes once exposed to PPARγ activator like EPA and DHA that are highly suggested during pregnancy. The objective of this study aims at determining the identity of trans-differentiated cells by exploring the effect of EPA and DHA on C2C12 undergoing differentiation into brown and white adipocytes. DHA but not EPA committed C2C12 cells reprograming into white like adipocyte phenotype. Also, DHA promoted the expression of lipolysis regulating genes but had no effect on genes regulating β-oxidation referring to its implication in lipid re-esterification. Furthermore, DHA impaired C2C12 cells differentiation into brown adipocytes through reducing the thermogenic capacity and mitochondrial biogenesis of derived cells independent of UCP1. Accordingly, DHA treated groups showed an increased accumulation of lipid droplets and suppressed mitochondrial maximal respiration and spare respiratory capacity. EPA, on the other hand, reduced myogenesis regulating genes, but no significant differences were observed in the expression of adipogenesis key genes. Likewise, EPA suppressed the expression of WAT signature genes indicating that EPA and DHA have an independent role on white adipogensis. Unlike DHA treatment, EPA supplementation had no effect on the differential of C2C12 cells into brown adipocytes. In conclusion, DHA is a potent adipogenic and lipogenic factor that can change the metabolic profile of muscle cells by increasing myocellular fat.

DOI: https://doi.org/10.1371/journal.pone.0249438
Exp Physiol. 2021 Sep 01.

The effects of pulmonary hypertension on skeletal muscle oxygen pressures in contracting rat spinotrapezius muscle.

Kiana M Schulze, Ramona E Weber, Trenton D Colburn, Andrew G Horn, Carl J Ade, Wei-Wen Hsu, David C Poole, Timothy I Musch.

   NEW FINDINGS: What is the central question of this study? This investigation was designed to determine whether impairment in the dynamics of O2 transport in skeletal muscle during a series of contractions might constitute a potential mechanism underlying reduced exercise capacity in pulmonary hypertension. What is the main finding and its importance? Our findings demonstrate that pulmonary hypertension compromises the dynamic matching of skeletal muscle O2 delivery-to-utilization following contractions onset in the rat spinotrapezius muscle. These results implicate a role for vascular dysfunction in the slow V̇O2 kinetics and exercise intolerance present in pulmonary hypertension.
ABSTRACT: Pulmonary hypertension (PH) is characterized by pulmonary vascular dysfunction and exercise intolerance due, in part, to compromised pulmonary and cardiac function. We tested the hypothesis that there are peripheral (i.e. skeletal muscle) aberrations in O2 delivery (Q̇O2 )-to-O2 utilization (V̇O2 ) matching and vascular control that might help to explain poor exercise tolerance in PH. Furthermore, we investigated the peripheral effects of nitric oxide (NO) in attenuating these decrements. Male Sprague-Dawley rats (n = 21) were administered monocrotaline (MCT; 50 mg/kg, i.p.) to induce PH. Disease progression was monitored via echocardiography. Phosphorescence quenching determined the O2 partial pressure in the interstitial space (PO2 is) in the spinotrapezius muscle at rest and during contractions under control (SNP-) and NO-donor (SNP+) conditions. MCT rats displayed RV hypertrophy (RV/(LV+S): 0.44(0.13) vs. 0.28(0.05)), pulmonary congestion, increased right ventricular systolic pressure (48(18) vs. 20(8) mmHg), and arterial hypoxaemia (PaO2 : 64(9) vs. 82(9) mmHg) compared to healthy controls (HC) (P<0.05). PO2 is was significantly lower in MCT rats during the first 30 seconds of SNP- contractions. SNP superfusion elevated PO2 is in both groups, however MCT rats demonstrated a lower PO2 is throughout SNP+ contractions versus HC (P<0.05). Thus, for small muscle mass exercise in MCT rats, muscle oxygenation is impaired across the rest-to-contractions transition and exogenous NO does not raise the Q̇O2 -to-V̇O2 ratio in contracting muscle to the same levels as HC. These data support muscle Q̇O2 -to-V̇O2 mismatch as a potential contributor to slow V̇O2 kinetics and therefore exercise intolerance in PH and suggest peripheral vascular dysfunction or remodeling as a possible mechanism. This article is protected by copyright. All rights reserved.

Keywords:  microvasculature; monocrotaline; nitric oxide; oxygen transport

DOI:  https://doi.org/10.1113/EP089631
Structure. 2021 Aug 26. pii: S0969-2126(21)00296-3. [Epub ahead of print]

High-resolution structure of the membrane-embedded skeletal muscle ryanodine receptor.

Zephan Melville, Kookjoo Kim, Oliver B Clarke, Andrew R Marks.

  The type 1 ryanodine receptor (RyR)/calcium release channel on the sarcoplasmic reticulum (SR) is required for skeletal muscle excitation-contraction coupling and is the largest known ion channel, composed of four 565-kDa protomers. Cryogenic electron microscopy (cryo-EM) studies of the RyR have primarily used detergent to solubilize the channel; in the present study, we have used cryo-EM to solve high-resolution structures of the channel in liposomes using a gel-filtration approach with on-column detergent removal to form liposomes and incorporate the channel simultaneously. This allowed us to resolve the structure of the channel in the primed and open states at 3.4 and 4.0 Å, respectively, with a single dataset. This method offers validation for detergent-based structures of the RyR and offers a starting point for utilizing a chemical gradient mimicking the SR, where Ca2+ concentrations are millimolar in the lumen and nanomolar in the cytosol.

Keywords:  calcium release channel; cryoelectron microscopy; excitation-contraction coupling; ion channel; membrane protein; proteoliposomes; ryanodine receptor; sarcoplasmic reticulum; skeletal muscle

DOI:  https://doi.org/10.1016/j.str.2021.08.001
Proteins. 2021 Aug 29.

Mapping Co-regulatory Interactions among Ligand Binding sites in RyR1.

Venkat R Chirasani, Konstantin I Popov, Gerhard Meissner, Nikolay V Dokholyan.

  Ryanodine receptor 1 (RyR1) is an intracellular calcium ion (Ca2+ ) release channel required for skeletal muscle contraction. Although cryo-electron microscopy identified binding sites of three coactivators Ca2+ , ATP and caffeine (CFF), the mechanism of co-regulation and synergy of these activators is unknown. Here, we report allosteric connections among the three ligand binding sites and pore region in (i) Ca2+ bound-closed, (ii) ATP/CFF bound- closed, (iii) Ca2+ /ATP/CFF bound-closed, and (iv) Ca2+ /ATP/CFF bound-open RyR1 states. We identified two dominant networks of interactions that mediate communication between the Ca2+ binding site and pore region in Ca2+ bound-closed state, which partially overlapped with the pore communications in ATP/CFF bound-closed RyR1 state. In Ca2+ /ATP/CFF bound-closed and -open RyR1 states, co-regulatory interactions were analogous to communications in the Ca2+ bound-closed and ATP/CFF bound- closed states. Both ATP- and CFF- binding sites mediate communication between the Ca2+ binding site and the pore region in Ca2+ /ATP/CFF bound - open RyR1 structure. We conclude that Ca2+ , ATP, and CFF propagate their effects to the pore region through a network of overlapping interactions that mediate allosteric control and molecular synergy in channel regulation. This article is protected by copyright. All rights reserved.

Keywords:  Ryanodine receptor; allosteric coupling; and allosteric interactions; computational analysis; percolation theory; single channel measurements

DOI:  https://doi.org/10.1002/prot.26228
Eur J Pharmacol. 2021 Aug 27. pii: S0014-2999(21)00605-1. [Epub ahead of print]910 174451

Acetyl-CoA carboxylase 2 inhibition reduces skeletal muscle bioactive lipid content and attenuates progression of type 2 diabetes in Zucker diabetic fatty rats.

Hiroyuki Takagi, Tatsuya Ikehara, Kumi Hashimoto, Keiichi Tanimoto, Atsuyuki Shimazaki, Yuto Kashiwagi, Shingo Sakamoto, Hideo Yukioka.

  Intramyocellular lipid (IMCL) accumulation in skeletal muscle is closely associated with development of insulin resistance. In particular, diacylglycerol and ceramide are currently considered as causal bioactive lipids for impaired insulin action. Recently, inhibition of acetyl-CoA carboxylase 2 (ACC2), which negatively modulates mitochondrial fatty acid oxidation, has been shown to reduce total IMCL content and improve whole-body insulin resistance. This study aimed to investigate whether ACC2 inhibition-induced compositional changes in bioactive lipids, especially diacylglycerol and ceramide, within skeletal muscle contribute to the improved insulin resistance. In skeletal muscle of normal rats, treatment of the ACC2 inhibitor compound 2e significantly decreased both diacylglycerol and ceramide levels while having no significant impact on other lipid metabolite levels. In skeletal muscle of Zucker diabetic fatty (ZDF) rats, which exhibited greater lipid accumulation than that of normal rats, compound 2e significantly decreased diacylglycerol and ceramide levels corresponding to reduced long chain acyl-CoA pools. Additionally, in the lipid metabolomics study, ZDF rats treated with compound 2e also showed improved diabetes-related metabolic disturbance, as reflected by delayed hyperinsulinemia as well as upregulated gene expression associated with diabetic conditions in skeletal muscle. These metabolic improvements were strongly correlated with the bioactive lipid reductions. Furthermore, long-term treatment of compound 2e markedly improved whole-body insulin resistance, attenuated hyperglycemia and delayed insulin secretion defect even at severe diabetic conditions. These findings suggest that ACC2 inhibition decreases diacylglycerol and ceramide accumulation within skeletal muscle by enhancing acyl-CoA breakdown, leading to attenuation of lipid-induced insulin resistance and subsequent diabetes progression.

Keywords:  Acetyl-CoA carboxylase 2; Diabetes; Insulin resistance; Lipid accumulation; Skeletal muscle

DOI:  https://doi.org/10.1016/j.ejphar.2021.174451
J Virol. 2021 Sep 01. JVI0090421

Skeletal muscle is an early site of Zika virus replication and injury impairing myogenesis.

Daniel Gavino-Leopoldino, Camila Menezes Figueiredo, Mariana Oliveira Lopes da Silva, Letícia Gonçalves Barcellos, Rômulo Leão Silva Neris, Laryssa Daniele Miranda Pinto, Suzana Maria Bernardino Araújo, Leandro Ladislau, Claudia Farias Benjamim, Andrea T Da Poian, Julia Rosauro Clarke, Claudia Pinto Figueiredo, Iranaia Assunção-Miranda.

Zika virus (ZIKV) infection became a worldwide concern due to its correlation with the development of microcephaly and other neurological disorders. ZIKV neurotropism is well characterized, but the role of peripheral viral amplification to brain infection remains unknown. Here we found that ZIKV replicates in human primary skeletal muscle myoblasts, impairing its differentiation into myotubes but not interfering with the integrity of the already formed muscle fibers. Using mouse models, we showed ZIKV tropism to muscle tissue either during embryogenesis after maternal transmission or when infection occurred after birth. Interestingly, ZIKV replication in the mouse skeletal muscle started immediately after ZIKV inoculation, preceding viral RNA detection in the brain and causing no disruption to the integrity of the blood brain barrier, and remained active for more than two weeks, while replication in spleen and liver were not sustained over time. In addition, ZIKV infection of the skeletal muscle induces necrotic lesions, inflammation and fiber atrophy. We also found a reduction in the expression of regulatory myogenic factors that are essential for muscle repair after injury. Taken together our results indicate that the skeletal muscle is an early site of viral amplification and lesion that may result in late consequences in muscle development after ZIKV infection. Importance Zika Virus (ZIKV) neurotropism and its deleterious effects on central nervous system have been well characterized. But, investigations of the initial replication sites for the establishment of infection and viral spread to neural tissues remain under explored. The complete description of the range of ZIKV induced lesions and others factors that can influence the severity of the disease are necessary to prevent ZIKV deleterious effects. ZIKV has been shown to access the central nervous system without significantly affecting blood-brain barrier permeability. Here we demonstrated that skeletal muscle is an earlier site of ZIKV replication, contributing to the increase of peripheral ZIKV load. ZIKV replication in muscle promotes necrotic lesions, inflammation and also impairs myogenesis. Overall, our findings showed that skeletal muscle is involved in pathogenesis and opens new fields in the investigation of the long-term consequence of early infection.

DOI: https://doi.org/10.1128/JVI.00904-21
Sports Med Health Sci. 2020 Dec;2(4): 175-176

Emerging mechanisms for exercise effects on muscle wasting and anabolic resistance.

J William Deaver, Nicholas P Greene.

DOI: https://doi.org/10.1016/j.smhs.2020.11.001
J Vis Exp. 2021 Aug 14.

Dissection of Single Skeletal Muscle Fibers for Immunofluorescent and Morphometric Analyses of Whole-Mount Neuromuscular Junctions.

Carmen Bolatto, Silvia Olivera-Bravo, Sofía Cerri.

The neuromuscular junction (NMJ) is a specialized point of contact between the motor nerve and the skeletal muscle. This peripheral synapse exhibits high morphological and functional plasticity. In numerous nervous system disorders, NMJ is an early pathological target resulting in neurotransmission failure, weakness, atrophy, and even in muscle fiber death. Due to its relevance, the possibility to quantitatively assess certain aspects of the relationship between NMJ components can help to understand the processes associated with its assembly/disassembly. The first obstacle when working with muscles is to gain the technical expertise to quickly identify and dissect without damaging their fibers. The second challenge is to utilize high-quality detection methods to obtain NMJ images that can be used to perform quantitative analysis. This article presents a step-by-step protocol for dissecting extensor digitorum longus and soleus muscles from rats. It also explains the use of immunofluorescence to visualize pre and postsynaptic elements of whole-mount NMJs. Results obtained demonstrate that this technique can be used to establish the microscopic anatomy of the synapsis and identify subtle changes in the status of some of its components under physiological or pathological conditions.

DOI: https://doi.org/10.3791/62620
Am J Clin Nutr. 2021 Sep 02. pii: nqab283. [Epub ahead of print]

Can increasing physical activity prevent aging-related loss of skeletal muscle?

Steven B Heymsfield, Nicole Fearnbach.

DOI: https://doi.org/10.1093/ajcn/nqab283
Biomaterials. 2021 Aug 20. pii: S0142-9612(21)00428-2. [Epub ahead of print]277 121072

Minimally invasive co-injection of modular micro-muscular and micro-vascular tissues improves in situ skeletal muscle regeneration.

Ying Wang, Ranjith Kumar Kankala, Yuan-Yuan Cai, Han-Xiao Tang, Kai Zhu, Jian-Ting Zhang, Da-Yun Yang, Shi-Bin Wang, Yu Shrike Zhang, Ai-Zheng Chen.

  Various conventional treatment strategies for volumetric muscle loss (VML) are often hampered by the extreme donor site morbidity, the limited availability of quality muscle flaps, and complicated, as well as invasive surgical procedures. The conventional biomaterial-based scaffolding systems carrying myoblasts have been extensively investigated towards improving the regeneration of the injured muscle tissues, as well as their injectable forms. However, the applicability of such designed systems has been restricted due to the lack of available vascular networks. Considering these facts, here we present the development of a unique set of two minimally invasively injectable modular microtissues, consisting of mouse myoblast (C2C12)-laden poly(lactic-co-glycolic acid) porous microspheres (PLGA PMs), or the micro-muscles, and human umbilical vein endothelial cell (HUVEC)-laden poly(ethylene glycol) hollow microrods (PEG HMs), or the microvessels. Besides systematic in vitro investigations, the myogenic performance of these modular composite microtissues, when co-injected, was explored in vivo using a mouse VML model, which confirmed improved in situ muscle regeneration and remolding. Together, we believe that the construction of these injectable modular microtissues and their combination for minimally invasive therapy provides a promising method for in situ tissue healing.

Keywords:  Hollow microrods; Injectable scaffolds; Minimally invasive surgery; Vascularization; Volumetric muscle loss

DOI:  https://doi.org/10.1016/j.biomaterials.2021.121072
Mol Genet Metab. 2021 Aug 17. pii: S1096-7192(21)00767-8. [Epub ahead of print]

Muscle-directed AAV gene therapy rescues the maple syrup urine disease phenotype in a mouse model.

Jenny A Greig, Matthew Jennis, Aditya Dandekar, Joanna K Chorazeczewski, Melanie K Smith, Scott N Ashley, Hanying Yan, James M Wilson.

  Maple syrup urine disease (MSUD) is a rare, inherited metabolic disorder characterized by a dysfunctional mitochondrial enzyme complex, branched-chain alpha-keto acid dehydrogenase (BCKDH), which catabolizes branched-chain amino acids (BCAAs). Without functional BCKDH, BCAAs and their neurotoxic alpha-keto intermediates can accumulate in the blood and tissues. MSUD is currently incurable and treatment is limited to dietary restriction or liver transplantation, meaning there is a great need to develop new treatments for MSUD. We evaluated potential gene therapy applications for MSUD in the intermediate MSUD (iMSUD) mouse model, which harbors a mutation in the dihydrolipoamide branched-chain transacylase E2 (DBT) subunit of BCKDH. Systemic delivery of an adeno-associated virus (AAV) vector expressing DBT under control of the liver-specific TBG promoter to the liver did not sufficiently ameliorate all aspects of the disease phenotype. These findings necessitated an alternative therapeutic strategy. Muscle makes a larger contribution to BCAA metabolism than liver in humans, but a muscle-specific approach involving a muscle-specific promoter for DBT expression delivered via intramuscular (IM) administration only partially rescued the MSUD phenotype in mice. Combining the muscle-tropic AAV9 capsid with the ubiquitous CB7 promoter via IM or IV injection, however, substantially increased survival across all assessed doses. Additionally, near-normal serum BCAA levels were achieved and maintained in the mid- and high-dose cohorts throughout the study; this approach also protected these mice from a lethal high-protein diet challenge. Therefore, administration of a gene therapy vector that expresses in both muscle and liver may represent a viable approach to treating patients with MSUD.

Keywords:  Animal model; Branched-chain amino acids; Gene therapy; High protein diet; Intramuscular; Maple syrup urine disease

DOI:  https://doi.org/10.1016/j.ymgme.2021.08.003
Metabolism. 2021 Aug 31. pii: S0026-0495(21)00173-6. [Epub ahead of print] 154873

Diet and exercise reduce pre-existing NASH and fibrosis and have additional beneficial effects on the vasculature, adipose tissue and skeletal muscle via organ-crosstalk.

Anita M van den Hoek, Jelle C B C de Jong, Nicole Worms, Anita van Nieuwkoop, Marijke Voskuilen, Aswin L Menke, Serene Lek, Martien P M Caspers, Lars Verschuren, Robert Kleemann.

   BACKGROUND: Non-alcoholic steatohepatitis (NASH) has become one of the most common liver diseases and is still without approved pharmacotherapy. Lifestyle interventions using exercise and diet change remain the current treatment of choice and even a small weight loss (5-7%) can already have a beneficial effect on NASH. However, the underlying molecular mechanisms of exercise and diet interventions remain largely elusive, and it is unclear whether they exert their health effects via similar or different pathways.
METHODS: Ldlr-/-.Leiden mice received a high fat diet (HFD) for 30 weeks to establish a severe state of NASH/fibrosis with simultaneous atherosclerosis development. Groups of mice were then either left untreated (control group) or were treated for 20 weeks with exercise (running wheel), diet change (switch to a low fat chow diet) or the combination thereof. The liver and distant organs including heart, white adipose tissue (WAT) and muscle were histologically examined. Comprehensive transcriptome analysis of liver, WAT and muscle revealed the organ-specific effects of exercise and diet and defined the underlying pathways.
RESULTS: Exercise and dietary change significantly reduced body weight, fat mass, adipocyte size and improved myosteatosis and muscle function with additive effects of combination treatment. WAT inflammation was significantly improved by diet change, tended to be reduced with exercise, and combination therapy had no additive effect. Hepatic steatosis and inflammation were almost fully reversed by exercise and diet change, while hepatic fibrosis tended to be improved with exercise and was significantly improved with diet change. Additive effects for the combination therapy were shown for liver steatosis and associated liver lipids, and atherosclerosis, but not for hepatic inflammation and fibrosis. Pathway analysis revealed complementary effects on metabolic pathways and lipid handling processes, thereby substantiating the added value of combined lifestyle treatment.
CONCLUSIONS: Exercise, diet change and the combination thereof can reverse established NASH/fibrosis in obese Ldlr-/-.Leiden mice. In addition, the lifestyle interventions had beneficial effects on atherosclerosis, WAT inflammation and muscle function. For steatosis and other parameters related to adiposity or lipid metabolism, exercise and dietary change affected more distinct pathways that acted complementary when the interventions were combined resulting in an additive effect for the combination therapy on important endpoints including NASH and atherosclerosis. For inflammation, exercise and diet change shared several underlying pathways resulting in a net similar effect when the interventions were combined.

Keywords:  Atherosclerosis; Fibrosis; Lifestyle interventions; Muscle-liver axis; NAFLD; WAT-liver axis

DOI:  https://doi.org/10.1016/j.metabol.2021.154873
Exp Physiol. 2021 Sep 01.

Physiological complexity: Influence of ageing, disease and neuromuscular fatigue on muscle force and torque fluctuations.

Jamie Pethick, Samantha L Winter, Mark Burnley.

   NEW FINDINGS: What is the topic of this review? We review physiological complexity in muscle force and torque fluctuations; specifically, we focus on the quantification of complexity, how neuromuscular complexity is altered by perturbations and the potential mechanism underlying changes in neuromuscular complexity. What advances does it highlight? We highlight the necessity to calculate both magnitude- and complexity-based measures for the thorough evaluation of force/torque fluctuations. We also highlight the need for further research on neuromuscular complexity, particularly how it relates to the performance of functional activities (e.g. manual dexterity, balance, locomotion).
ABSTRACT: Physiological time-series produce inherently complex fluctuations. In the last 30 years, methods have been developed to characterise these fluctuations, and have revealed that such fluctuations contain information about the function of the system producing them. Two broad classes of metrics are used: 1) those which quantify the regularity of the signal (e.g. entropy metrics); and 2) those which quantify the fractal properties of the signal (e.g. detrended fluctuation analysis). Using these techniques, it has been demonstrated that aging results in a loss of complexity in the time-series of a multitude of signals, including heart rate, respiration, gait and, crucially, muscle force or torque output. This suggests that as the body ages, physiological systems become less adaptable (i.e. the systems' ability to respond rapidly to a changing external environment is diminished). More recently, it has been shown that neuromuscular fatigue causes a substantial loss of muscle torque complexity, a process that can be observed in a few minutes, rather than the decades it requires the same system to degrade with aging. The loss of torque complexity with neuromuscular fatigue appears to occur exclusively above the critical torque (at least for tasks lasting up to 30 minutes). The loss of torque complexity can be exacerbated with previous exercise of the same limb, and reduced by the administration of caffeine, suggesting both peripheral and central mechanisms contribute to this loss. The mechanisms underpinning the loss of complexity are not known but may be related to altered motor unit behaviour as the muscle fatigues. This article is protected by copyright. All rights reserved.

Keywords:  ageing; complexity; fractal; muscle force/torque; neuromuscular fatigue

DOI:  https://doi.org/10.1113/EP089711
Life Sci Alliance. 2021 Nov;pii: e202101034. [Epub ahead of print]4(11):

High levels of TFAM repress mammalian mitochondrial DNA transcription in vivo.

Nina A Bonekamp, Min Jiang, Elisa Motori, Rodolfo Garcia Villegas, Camilla Koolmeister, Ilian Atanassov, Andrea Mesaros, Chan Bae Park, Nils-Göran Larsson.

Mitochondrial transcription factor A (TFAM) is compacting mitochondrial DNA (dmtDNA) into nucleoids and directly controls mtDNA copy number. Here, we show that the TFAM-to-mtDNA ratio is critical for maintaining normal mtDNA expression in different mouse tissues. Moderately increased TFAM protein levels increase mtDNA copy number but a normal TFAM-to-mtDNA ratio is maintained resulting in unaltered mtDNA expression and normal whole animal metabolism. Mice ubiquitously expressing very high TFAM levels develop pathology leading to deficient oxidative phosphorylation (OXPHOS) and early postnatal lethality. The TFAM-to-mtDNA ratio varies widely between tissues in these mice and is very high in skeletal muscle leading to strong repression of mtDNA expression and OXPHOS deficiency. In the heart, increased mtDNA copy number results in a near normal TFAM-to-mtDNA ratio and maintained OXPHOS capacity. In liver, induction of LONP1 protease and mitochondrial RNA polymerase expression counteracts the silencing effect of high TFAM levels. TFAM thus acts as a general repressor of mtDNA expression and this effect can be counterbalanced by tissue-specific expression of regulatory factors.

DOI: https://doi.org/10.26508/lsa.202101034
Proc Natl Acad Sci U S A. 2021 Sep 07. pii: e2105951118. [Epub ahead of print]118(36):

RNA-bound PGC-1α controls gene expression in liquid-like nuclear condensates.

Joaquín Pérez-Schindler, Bastian Kohl, Konstantin Schneider-Heieck, Aurel B Leuchtmann, Carlos Henríquez-Olguín, Volkan Adak, Geraldine Maier, Julien Delezie, Thomas Sakoparnig, Elyzabeth Vargas-Fernández, Bettina Karrer-Cardel, Danilo Ritz, Alexander Schmidt, Maria Hondele, Thomas E Jensen, Sebastian Hiller, Christoph Handschin.

  Plasticity of cells, tissues, and organs is controlled by the coordinated transcription of biological programs. However, the mechanisms orchestrating such context-specific transcriptional networks mediated by the dynamic interplay of transcription factors and coregulators are poorly understood. The peroxisome proliferator-activated receptor γ coactivator 1α (PGC-1α) is a prototypical master regulator of adaptive transcription in various cell types. We now uncovered a central function of the C-terminal domain of PGC-1α to bind RNAs and assemble multiprotein complexes including proteins that control gene transcription and RNA processing. These interactions are important for PGC-1α recruitment to chromatin in transcriptionally active liquid-like nuclear condensates. Notably, such a compartmentalization of active transcription mediated by liquid-liquid phase separation was observed in mouse and human skeletal muscle, revealing a mechanism by which PGC-1α regulates complex transcriptional networks. These findings provide a broad conceptual framework for context-dependent transcriptional control of phenotypic adaptations in metabolically active tissues.

Keywords:  RNA-binding protein; chromatin; gene transcription; liquid–liquid phase separation; transcriptional coactivator

DOI:  https://doi.org/10.1073/pnas.2105951118
Elife. 2021 09 02. pii: e67182. [Epub ahead of print]10

Muscle-specific economy of force generation and efficiency of work production during human running.

Sebastian Bohm, Falk Mersmann, Alessandro Santuz, Arno Schroll, Adamantios Arampatzis.

  Human running features a spring-like interaction of body and ground, enabled by elastic tendons that store mechanical energy and facilitate muscle operating conditions to minimize the metabolic cost. By experimentally assessing the operating conditions of two important muscles for running, the soleus and vastus lateralis, we investigated physiological mechanisms of muscle work production and muscle force generation. We found that the soleus continuously shortened throughout the stance phase, operating as work generator under conditions that are considered optimal for work production: high force-length potential and high enthalpy efficiency. The vastus lateralis promoted tendon energy storage and contracted nearly isometrically close to optimal length, resulting in a high force-length-velocity potential beneficial for economical force generation. The favorable operating conditions of both muscles were a result of an effective length and velocity-decoupling of fascicles and muscle-tendon unit, mostly due to tendon compliance and, in the soleus, marginally by fascicle rotation.

Keywords:  enthalpy-velocity relationship; force-length and force-velocity relationship; human; length- and velocity-decoupling; metabolic cost of running; physics of living systems; soleus muscle; tendon elasticity; vastus lateralis muscle

DOI:  https://doi.org/10.7554/eLife.67182
Cell Discov. 2020 Sep 01. 6(1): 59

RGS12 is required for the maintenance of mitochondrial function during skeletal development.

Gongsheng Yuan, Shuting Yang, Min Liu, Shuying Yang.

Mitochondrial morphology and function are crucial for tissue homeostasis, such as for skeletal development, but the cellular and molecular mechanisms remain unclear. Here, we provide evidence that regulator of G-protein signaling 12 (RGS12) is present in the mitochondria of primary chondrocytes and cartilage tissues. Deletion of RGS12 in type II collagen-positive cells led to a significant decrease in mitochondrial number, membrane potential, and oxidative phosphorylation function. Mechanistically, RGS12 promoted the function of ATP5A as an enhancer of tyrosine phosphorylation. Mice with RGS12 deficiency in the chondrocyte lineage showed serious body retardation, decreased bone mass, and chondrocyte apoptosis due to the defective activity of ATP synthase. To our knowledge, this is the first report that RGS12 is required for maintaining the function of mitochondria, which may allow it to orchestrate responses to cellular homeostasis.

DOI: https://doi.org/10.1038/s41421-020-00190-w