bims-moremu 2021-08-22 papers

bims-moremu

Biomed News

on Molecular regulators of muscle mass

Issue of 2021–08–22
33 papers selected by
Anna Vainshtein, Craft Science Inc.

Lifelong Exercise in Age Rats Improves Skeletal Muscle Function and MicroRNA Profile.
Chromatin accessibility profiling identifies evolutionary conserved loci in activated human satellite cells.
AMP deamination is sufficient to replicate an atrophy-like metabolic phenotype in skeletal muscle.
Metformin and leucine increase satellite cells and collagen remodeling during disuse and recovery in aged muscle.
Ketogenic diet with medium-chain triglycerides restores skeletal muscle function and pathology in a rat model of Duchenne muscular dystrophy.
Raising the Guanosine-Based Molecules as Regulators of Excitable Tissues by the Exosomal-Vehiculated Signaling.
Electrophysiological analysis of healthy and dystrophic 3D bioengineered skeletal muscle tissues.
Prolonged caloric restriction ameliorates age-related atrophy in slow and fast muscle fibers of rat soleus muscle.
Lack of Increase in Muscle Mitochondrial Protein Synthesis During the Course of Aerobic Exercise and Its Recovery in the Fasting State Irrespective of Obesity.
The Effect of Mechanical Stretch on Myotube Growth Suppression by Colon-26 Tumor-Derived Factors.
Activation of YAP regulates muscle fiber size in a PKC-dependent mechanism during chick in vitro myogenesis.
Skeletal muscle mitochondrial network dynamics in metabolic disorders and aging.
Isolation of human fibroadipogenic progenitors and satellite cells from frozen muscle biopsies.
A xanthene derivative, DS20060511, attenuates glucose intolerance by inducing skeletal muscle-specific GLUT4 translocation in mice.
A Physiology-Guided Classification of Active-Stress and Active-Strain Approaches for Continuum-Mechanical Modeling of Skeletal Muscle Tissue.
Molecular and Biomechanical Adaptations to Mechanical Stretch in Cultured Myotubes.
Effect of Local Heat Application During Exercise on Gene Expression Related to Mitochondrial Homeostasis.
CREB-mediated transcriptional activation of NRMT1 drives muscle differentiation.
Myeloid cell-mediated targeting of LIF to dystrophic muscle causes transient increases in muscle fiber lesions by disrupting the recruitment and dispersion of macrophages in muscle.
Cold-Induced RNA-Binding Protein Promotes Glucose Metabolism and Reduces Apoptosis by Increasing AKT Phosphorylation in Mouse Skeletal Muscle Under Acute Cold Exposure.
Cell-type selective targeted delivery of a recombinant lysosomal enzyme for enzyme therapies.
Translational control by DHX36 binding to 5'UTR G-quadruplex is essential for muscle stem-cell regenerative functions.
Branched-chain amino acids at supraphysiological but not physiological levels reduce myotube insulin sensitivity.
Extracellular miRNAs as mediators of obesity-associated disease.
A mutation in desmin makes skeletal muscle less vulnerable to acute muscle damage after eccentric loading in rats.
Dissecting dual roles of MyoD during lineage conversion to mature myocytes and myogenic stem cells.
Normal to enhanced intrinsic mitochondrial respiration in skeletal muscle of middle- to older-aged women and men with uncomplicated type 1 diabetes.
Nox4 Knockout Does Not Prevent Diaphragm Atrophy, Contractile Dysfunction, or Mitochondrial Maladaptation in the Early Phase Post-Myocardial Infarction in Mice.
Filamin C Cardiomyopathy Variants Cause Protein and Lysosome Accumulation.
Myogenic Cell Transplantation in Genetic and Acquired Diseases of Skeletal Muscle.
Muscle-specific sirtuin 3 overexpression does not attenuate the pathological effects of high-fat/high-sucrose feeding but does enhance cardiac SERCA2a activity.
The effects of cocoa flavanols on indices of muscle recovery and exercise performance: a narrative review.
Titin kinase ubiquitination aligns autophagy receptors with mechanical signals in the sarcomere.

Med Sci Sports Exerc. 2021 Sep 01. 53(9): 1873-1882

Lifelong Exercise in Age Rats Improves Skeletal Muscle Function and MicroRNA Profile.

Hao-En Gao, Fang-Hui Li, Tian Xie, Song Ma, Yi-Bo Qiao, DA-Shuai Wu, Lei Sun.

PURPOSE: Lifelong exercise is known to attenuate sarcopenia (age-associated reduction in muscle mass and function); however, the underlying molecular mechanisms remain unclear. As microRNAs are widely involved in the regulation of skeletal muscle growth and development, we aimed to evaluate the effects of lifelong regular exercise on age-related alterations in muscle microRNA expression profiles as well as on skeletal muscle atrophy, apoptosis, and mitochondria and autophagy dysfunction.
METHODS: Female 8-month-old Sprague-Dawley rats were divided into four groups; 1) 18 months of moderate-intensity continuous training (MICT) initiated at 8 months (adult-MICT, n = 12), 2) 8 months of MICT initiated at 18 months (presarcopenia-MICT, n = 12), 3) 8-month-old adult sedentary controls (adult-SED), and 4) 26-month-old aging sedentary controls (old-SED). Age skeletal muscles were then subjected to quantitative reverse transcription-polymerase chain reaction, Kyoto Encyclopedia of Genes and Genomes, immunoblotting, and miR-486 3' untranslated region luciferase reporter gene analyses.
RESULTS: Age-related loss of miR-486 expression was improved, skeletal muscle atrophy and apoptosis were downregulated, and mitochondrial activity and autophagy were upregulated in the adult-MICT group. Kyoto Encyclopedia of Genes and Genomes analysis revealed that the PI3K/Akt pathway was upregulated in adult-MICT rats compared with that in old-SED. In vitro analyses in rat skeletal muscle L6 cells further confirmed that miR-486 targets PTEN, not SAV1, thereby activating the PI3K/Akt pathway and indirectly inhibiting HIPPO signaling.
CONCLUSIONS: Compared with presarcopenia-MICT rats, adult-MICT rats experienced greater beneficial effects regarding ameliorated age-related alterations in muscle miRNA expression profile, skeletal muscle atrophy, apoptosis, and mitochondria and autophagy dysfunction, which is potentially associated with the increased miR-486 expression and concomitant targeting of the PTEN/Akt signaling pathway.

DOI: https://doi.org/10.1249/MSS.0000000000002661
Stem Cell Res. 2021 Aug 11. pii: S1873-5061(21)00343-3. [Epub ahead of print]55 102496

Chromatin accessibility profiling identifies evolutionary conserved loci in activated human satellite cells.

Lisa S Chow, Darko Bosnakovski, Douglas G Mashek, Michael Kyba, Rita C R Perlingeiro, Alessandro Magli.

  Satellite cells represent the main myogenic population accounting for skeletal muscle homeostasis and regeneration. While our knowledge of the signaling pathways controlling satellite cell regenerative capability is increasing, the underlying epigenetic mechanisms are still not clear, especially in the case of human satellite cells. Here, by performing chromatin accessibility profiling (ATAC-seq) in samples isolated from human and murine muscles, we investigated the changes in the epigenetic landscape occurring during the transition from activated satellite cells to myoblasts. Our analysis identifies a compendium of putative regulatory elements defining human activated satellite cells and myoblasts, respectively. A subset of these differentially accessible loci is shared by both murine and human satellite cells, includes elements associated with known self-renewal regulators, and is enriched for motifs bound by transcription factors participating in satellite cell regulation. Integration of transcriptional and epigenetic data reveals that known regulators of metabolic gene expression, such as PPARGC1A, represent potential PAX7 targets. Through characterization of genomic networks and the underlying effectors, our data represent an important starting point for decoding and manipulating the molecular mechanisms underlying human satellite cell muscle regenerative potential.

Keywords:  ATAC-seq; Human satellite cell; Muscle stem cell; PGC1 alpha; Pax7; Regulatory element

DOI:  https://doi.org/10.1016/j.scr.2021.102496
Metabolism. 2021 Aug 13. pii: S0026-0495(21)00164-5. [Epub ahead of print] 154864

AMP deamination is sufficient to replicate an atrophy-like metabolic phenotype in skeletal muscle.

Spencer G Miller, Paul S Hafen, Andrew S Law, Catherine B Springer, David L Logsdon, Thomas M O'Connell, Carol A Witczak, Jeffrey J Brault.

   BACKGROUND: Skeletal muscle atrophy, whether caused by chronic disease, acute critical illness, disuse or aging, is characterized by tissue-specific decrease in oxidative capacity and broad alterations in metabolism that contribute to functional decline. However, the underlying mechanisms responsible for these metabolic changes are largely unknown. One of the most highly upregulated genes in atrophic muscle is AMP deaminase 3 (AMPD3: AMP → IMP+NH3), which controls the content of intracellular adenine nucleotides (AdN; ATP + ADP + AMP). Given the central role of AdN in signaling mitochondrial gene expression and directly regulating metabolism, we hypothesized that overexpressing AMPD3 in muscle cells would be sufficient to alter their metabolic phenotype similar to that of atrophic muscle.
METHODS: AMPD3 and GFP (control) were overexpressed in mouse tibialis anterior (TA) muscles via plasmid electroporation and in C2C12 myotubes using adenovirus vectors. TA muscles were excised one week later, and AdN were quantified by UPLC. In myotubes, targeted measures of AdN, AMPK/PGC-1α/mitochondrial protein synthesis rates, unbiased metabolomics, and transcriptomics by RNA sequencing were measured after 24 h of AMPD3 overexpression. Media metabolites were measured as an indicator of net metabolic flux. At 48 h, the AMPK/PGC-1α/mitochondrial protein synthesis rates, and myotube respiratory function/capacity were measured.
RESULTS: TA muscles overexpressing AMPD3 had significantly less ATP than contralateral controls (-25%). In myotubes, increasing AMPD3 expression for 24 h was sufficient to significantly decrease ATP concentrations (-16%), increase IMP, and increase efflux of IMP catabolites into the culture media, without decreasing the ATP/ADP or ATP/AMP ratios. When myotubes were treated with dinitrophenol (mitochondrial uncoupler), AMPD3 overexpression blunted decreases in ATP/ADP and ATP/AMP ratios but exacerbated AdN degradation. As such, pAMPK/AMPK, pACC/ACC, and phosphorylation of AMPK substrates, were unchanged by AMPD3 at this timepoint. AMPD3 significantly altered 191 out of 639 detected intracellular metabolites, but only 30 transcripts, none of which encoded metabolic enzymes. The most altered metabolites were those within purine nucleotide, BCAA, glycolysis, and ceramide metabolic pathways. After 48 h, AMPD3 overexpression significantly reduced pAMPK/AMPK (-24%), phosphorylation of AMPK substrates (-14%), and PGC-1α protein (-22%). Moreover, AMPD3 significantly reduced myotube mitochondrial protein synthesis rates (-55%), basal ATP synthase-dependent (-13%), and maximal uncoupled oxygen consumption (-15%).
CONCLUSIONS: Increased expression of AMPD3 significantly decreased mitochondrial protein synthesis rates and broadly altered cellular metabolites in a manner similar to that of atrophic muscle. Importantly, the changes in metabolites occurred prior to reductions in AMPK signaling, gene expression, and mitochondrial protein synthesis, suggesting metabolism is not dependent on reductions in oxidative capacity, but may be consequence of increased AMP deamination. Therefore, AMP deamination in skeletal muscle may be a mechanism that alters the metabolic phenotype of skeletal muscle during atrophy and could be a target to improve muscle function during muscle wasting.

Keywords:  AMP activated protein kinase; AMP deaminase; ATP; Metabolomics; Mitochondrial biogenesis; Muscle atrophy

DOI:  https://doi.org/10.1016/j.metabol.2021.154864
FASEB J. 2021 Sep;35(9): e21862

Metformin and leucine increase satellite cells and collagen remodeling during disuse and recovery in aged muscle.

Jonathan J Petrocelli, Ziad S Mahmassani, Dennis K Fix, Jessie A Montgomery, Paul T Reidy, Alec I McKenzie, Naomi M de Hart, Patrick J Ferrara, Joshua J Kelley, Hiroaki Eshima, Katsuhiko Funai, Micah J Drummond.

  Loss of muscle mass and strength after disuse followed by impaired muscle recovery commonly occurs with aging. Metformin (MET) and leucine (LEU) individually have shown positive effects in skeletal muscle during atrophy conditions but have not been evaluated in combination nor tested as a remedy to enhance muscle recovery following disuse atrophy in aging. The purpose of this study was to determine if a dual treatment of metformin and leucine (MET + LEU) would prevent disuse-induced atrophy and/or promote muscle recovery in aged mice and if these muscle responses correspond to changes in satellite cells and collagen remodeling. Aged mice (22-24 months) underwent 14 days of hindlimb unloading (HU) followed by 7 or 14 days of reloading (7 or 14 days RL). MET, LEU, or MET + LEU was administered via drinking water and were compared to Vehicle (standard drinking water) and ambulatory baseline. We observed that during HU, MET + LEU resolved whole body grip strength and soleus muscle specific force decrements caused by HU. Gastrocnemius satellite cell abundance was increased with MET + LEU treatment but did not alter muscle size during disuse or recovery conditions. Moreover, MET + LEU treatment alleviated gastrocnemius collagen accumulation caused by HU and increased collagen turnover during 7 and 14 days RL driven by a decrease in collagen IV content. Transcriptional pathway analysis revealed that MET + LEU altered muscle hallmark pathways related to inflammation and myogenesis during HU. Together, the dual treatment of MET and LEU was able to increase muscle function, satellite cell content, and reduce collagen accumulation, thus improving muscle quality during disuse and recovery in aging.

Keywords:  aging; extracellular matrix; fibrosis; hindlimb unloading; inflammation; reloading

DOI:  https://doi.org/10.1096/fj.202100883R
FASEB J. 2021 Sep;35(9): e21861

Ketogenic diet with medium-chain triglycerides restores skeletal muscle function and pathology in a rat model of Duchenne muscular dystrophy.

Yuri Fujikura, Hidetoshi Sugihara, Masaki Hatakeyama, Katsutaka Oishi, Keitaro Yamanouchi.

  Duchenne muscular dystrophy (DMD) is an intractable genetic disease associated with progressive skeletal muscle weakness and degeneration. Recently, it was reported that intraperitoneal injections of ketone bodies partially ameliorated muscular dystrophy by increasing satellite cell (SC) proliferation. Here, we evaluated whether a ketogenic diet (KD) with medium-chain triglycerides (MCT-KD) could alter genetically mutated DMD in model rats. We found that the MCT-KD significantly increased muscle strength and fiber diameter in these rats. The MCT-KD significantly suppressed the key features of DMD, namely, muscle necrosis, inflammation, and subsequent fibrosis. Immunocytochemical analysis revealed that the MCT-KD promoted the proliferation of muscle SCs, suggesting enhanced muscle regeneration. The muscle strength of DMD model rats fed with MCT-KD was significantly improved even at the age of 9 months. Our findings suggested that the MCT-KD ameliorates muscular dystrophy by inhibiting myonecrosis and promoting the proliferation of muscle SCs. As far as we can ascertain, this is the first study to apply a functional diet as therapy for DMD in experimental animals. Further studies are needed to elucidate the underlying mechanisms of the MCT-KD-induced improvement of DMD.

Keywords:  Duchenne muscular dystrophy; ketogenic diet; ketone bodies; nutrition therapy; skeletal muscle

DOI:  https://doi.org/10.1096/fj.202100629R
Front Pharmacol. 2021 ;12 658370

Raising the Guanosine-Based Molecules as Regulators of Excitable Tissues by the Exosomal-Vehiculated Signaling.

Tiziana Pietrangelo.



Keywords:  exosomes; guanosine; neopterin; satellite cells; skeletal muscle

DOI:  https://doi.org/10.3389/fphar.2021.658370
Am J Physiol Cell Physiol. 2021 08 18.

Electrophysiological analysis of healthy and dystrophic 3D bioengineered skeletal muscle tissues.

Christine T Nguyen, Majid Ebrahimi, Penney M Gilbert, Bryan Andrew Stewart.

  Recently, methods for creating three-dimensional (3D) human skeletal muscle tissues from myogenic cell lines have been reported. Bioengineered muscle tissues are contractile and respond to electrical and chemical stimulation. In this study we provide an electrophysiological analysis of healthy and dystrophic 3D bioengineered skeletal muscle tissues. We focus on Duchenne muscular dystrophy (DMD), a fatal muscle disorder involving the skeletal muscle system. The dystrophin gene, which when mutated causes DMD, encodes for the Dystrophin protein, which anchors the cytoskeletal network inside of a muscle cell to the extracellular matrix outside the cell. Here, we enlist a 3D in vitro model of DMD muscle tissue, to evaluate an understudied aspect of DMD, muscle cell electrical properties uncoupled from presynaptic neural inputs. Our data shows that electrophysiological aspects of DMD are replicated in the 3D bioengineered skeletal muscle tissue model. Furthermore, we test a block co-polymer, poloxamer 188, and demonstrate capacity for improving the membrane potential in DMD muscle. Therefore, this study serves as the baseline for a new in vitro method to examine potential therapies directed at muscular disorders.

Keywords:  DMD; Dystrophin; Electrophysiology; Human skeletal muscle; immortalized human myoblast

DOI:  https://doi.org/10.1152/ajpcell.00049.2021
Exp Gerontol. 2021 Aug 17. pii: S0531-5565(21)00301-6. [Epub ahead of print] 111519

Prolonged caloric restriction ameliorates age-related atrophy in slow and fast muscle fibers of rat soleus muscle.

Yuhei Mizunoe, Masaki Kobayashi, Hiroki Saito, Akifumi Goto, Ryota Migitaka, Kumi Miura, Naoyuki Okita, Yuka Sudo, Ryoma Tagawa, Miki Yoshida, Ai Umemori, Yoshimi Nakagawa, Hitoshi Shimano, Yoshikazu Higami.

  Aging causes loss of skeletal muscle mass and function, which is called sarcopenia. While sarcopenia impairs the quality of life of older adults and is a major factor in long-term hospitalization, its detailed pathogenic mechanism and preventive measures remain to be identified. Caloric restriction (CR) suppresses age-related physiological and pathological changes in many species and prolongs the average and healthy life expectancy. It has recently been reported that CR suppresses the onset of sarcopenia; however, few studies have analyzed the effects of long-term CR on age-related skeletal muscle atrophy. Thus, we investigated the aging and CR effects on soleus (SOL) muscles of 9-, 24-, and 29-month-old ad libitum-fed rats (9AL, 24AL, and 29AL, respectively) and of 29-month-old CR (29CR) rats. The total muscle cross sectional area (mCSA) of the entire SOL muscle significantly decreased in the 29AL rats, but not in the 24AL rats, compared with the 9AL rats. SOL muscle of the 29AL rats exhibited marked muscle fiber atrophy and increases in the number of muscle fibers with a central nucleus, in fibrosis, and in adipocyte infiltration. Additionally, although the decrease in the single muscle fiber cross-sectional area (fCSA) and the muscle fibers' number occurred in both slow-type and fast-type muscle fibers, the degree of atrophy was more remarkable in the fast-type fibers. However, CR suppressed the muscle fiber atrophy observed in the 29AL rats' SOL muscle by preserving the mCSA and the number of muscle fibers that declined with aging, and by decreasing the number of muscle fibers with a central nucleus, fibrosis and denervated muscle fibers. Overall, these results revealed that advanced aging separately reduces the number and fCSA of each muscle fiber type, but long-term CR can ameliorate this age-related sarcopenic muscle atrophy.

Keywords:  Aging; Caloric restriction; Muscle fiber type; Skeletal muscle

DOI:  https://doi.org/10.1016/j.exger.2021.111519
Front Physiol. 2021 ;12 702742

Lack of Increase in Muscle Mitochondrial Protein Synthesis During the Course of Aerobic Exercise and Its Recovery in the Fasting State Irrespective of Obesity.

Nathan Serrano, Lee Tran, Nyssa Hoffman, Lori Roust, Elena A De Filippis, Chad C Carroll, Shivam H Patel, Katon A Kras, Matthew Buras, Christos S Katsanos.

  Acute aerobic exercise induces skeletal muscle mitochondrial gene expression, which in turn can increase muscle mitochondrial protein synthesis. In this regard, the peroxisome proliferator-activated receptor γ coactivator 1α (PGC-1α), is a master regulator of mitochondrial biogenesis, and thus mitochondrial protein synthesis. However, PGC-1α expression is impaired in muscle of humans with obesity in response to acute aerobic exercise. Therefore, we sought to determine whether muscle mitochondrial protein synthesis is also impaired under the same conditions in humans with obesity. To this end, we measured mitochondrial and mixed-muscle protein synthesis in skeletal muscle of untrained subjects with (body fat: 34.7 ± 2.3%) and without (body fat: 25.3 ± 3.3%) obesity in a basal period and during a continuous period that included a 45 min cycling exercise (performed at an intensity corresponding to 65% of heart rate reserve) and a 3-h post-exercise recovery. Exercise increased PGC-1α mRNA expression in muscle of subjects without obesity, but not in subjects with obesity. However, muscle mitochondrial protein synthesis did not increase in either subject group. Similarly, mixed-muscle protein synthesis did not increase in either group. Concentrations of plasma amino acids decreased post-exercise in the subjects without obesity, but not in the subjects with obesity. We conclude that neither mitochondrial nor mixed-muscle protein synthesis increase in muscle of humans during the course of a session of aerobic exercise and its recovery period in the fasting state irrespective of obesity. Trial Registration: The study has been registered within ClinicalTrials.gov (NCT01824173).

Keywords:  PGC-1α; exercise; mitochondria; myosin heavy chain; protein synthesis

DOI:  https://doi.org/10.3389/fphys.2021.702742
Front Cell Dev Biol. 2021 ;9 690452

The Effect of Mechanical Stretch on Myotube Growth Suppression by Colon-26 Tumor-Derived Factors.

Jessica L Halle, Brittany R Counts-Franch, Rose M Prince, James A Carson.

  Preclinical models and in vitro experiments have provided valuable insight into the regulation of cancer-induced muscle wasting. Colon-26 (C26) tumor cells induce cachexia in mice, and conditioned media (CM) from these cells promotes myotube atrophy and catabolic signaling. While mechanical stimuli can prevent some effects of tumor-derived factors on myotubes, the impact of mechanical signaling on tumor-derived factor regulation of myosin heavy chain (MyHC) expression is not well understood. Therefore, we examined the effects of stretch-induced mechanical signaling on C2C12 myotube growth and MyHC expression after C26 CM exposure. C26 CM was administered to myotubes on day 5 of differentiation for 48 h. During the last 4 or 24 h of C26 CM exposure, 5% static uniaxial stretch was administered. C26 CM suppressed myotube growth and MyHC protein and mRNA expression. Stretch for 24 h increased myotube size and prevented the C26 CM suppression of MyHC-Fast protein expression. Stretch did not change suppressed MyHC mRNA expression. Stretch for 24 h reduced Atrogin-1/MAFbx, MuRF-1, and LC3B II/I ratio and increased integrin β1D protein expression and the myogenin-to-MyoD protein ratio. Stretch in the last 4 h of CM increased ERK1/2 phosphorylation but did not alter the CM induction of STAT3 or p38 phosphorylation. These results provide evidence that in myotubes pre-incubated with CM, the induction of mechanical signaling can still provide a growth stimulus and preserve MyHC-Fast protein expression independent of changes in mRNA expression.

Keywords:  cancer cachexia; myogenic differentiation; myosin heavy chain; passive stretch; skeletal muscle (myotubes)

DOI:  https://doi.org/10.3389/fcell.2021.690452
J Muscle Res Cell Motil. 2021 Aug 19.

Activation of YAP regulates muscle fiber size in a PKC-dependent mechanism during chick in vitro myogenesis.

Geyse Gomes, Kayo Moreira Bagri, Ivone de Andrade Rosa, Arnon Dias Jurberg, Claudia Mermelstein, Manoel Luis Costa.

  The formation of skeletal muscle fibers is an intricate process controlled by a multitude of signaling pathways, including Wnt, Shh, and FGF. However, the role of the Hippo pathway during vertebrate myofiber formation has conflicting reports, which we decided to address in chick muscle cultures. We found that the transcriptional regulator Yes-associated protein (YAP) was highly concentrated within the nuclei of myoblasts. As cells differentiate into myotubes, YAP localization shifted to the cell cytoplasm in more mature myotubes. Treatment of cultures with XMU-MP-1 (XMU), a MST1/2 inhibitor, stimulated the nuclear localization of YAP in myoblasts and in myotubes, upregulated myogenin, and promoted myoblast fusion, ultimately resulting in the formation of large and fully striated multinucleated myotubes. The XMU-induced phenotype was blocked by the protein kinase C (PKC) inhibitor calphostin, which raises the possibility that the Hippo pathway controls the growth of skeletal muscle fibers through a PKC-dependent mechanism.

Keywords:  Calphostin; Hippo pathway; Myogenesis; PKC; XMU-MP-1; YAP

DOI:  https://doi.org/10.1007/s10974-021-09608-8
Trends Mol Med. 2021 Aug 17. pii: S1471-4914(21)00198-2. [Epub ahead of print]

Skeletal muscle mitochondrial network dynamics in metabolic disorders and aging.

Ciarán E Fealy, Lotte Grevendonk, Joris Hoeks, Matthijs K C Hesselink.

  With global demographics trending towards an aging population, the numbers of individuals with an age-associated loss of independence is increasing. A key contributing factor is loss of skeletal muscle mitochondrial, metabolic, and contractile function. Recent advances in imaging technologies have demonstrated the importance of mitochondrial morphology and dynamics in the pathogenesis of disease. In this review, we examine the evidence for altered mitochondrial dynamics as a mechanism in age and obesity-associated loss of skeletal muscle function, with a particular focus on the available human data. We highlight some of the areas where more data are needed to identify the specific mechanisms connecting mitochondrial morphology and skeletal muscle dysfunction.

Keywords:  aging; metabolic disease; mitochondria; mitochondrial dynamics; sarcopenia

DOI:  https://doi.org/10.1016/j.molmed.2021.07.013
FASEB J. 2021 Sep;35(9): e21819

Isolation of human fibroadipogenic progenitors and satellite cells from frozen muscle biopsies.

Xavier Suárez-Calvet, Esther Fernández-Simón, Patricia Piñol-Jurado, Jorge Alonso-Pérez, Ana Carrasco-Rozas, Cinta Lleixà, Susana López-Fernández, Gemma Pons, Laura Soria, Anne Bigot, Vincent Mouly, Isabel Illa, Eduard Gallardo, Jyoti K Jaiswal, Jordi Díaz-Manera.

  Skeletal muscle contains multiple cell types that work together to maintain tissue homeostasis. Among these, satellite cells (SC) and fibroadipogenic progenitors cells (FAPs) are the two main stem cell pools. Studies of these cells using animal models have shown the importance of interactions between these cells in repair of healthy muscle, and degeneration of dystrophic muscle. Due to the unavailability of fresh patient muscle biopsies, similar analysis of interactions between human FAPs and SCs is limited especially among the muscular dystrophy patients. To address this issue here we describe a method that allows the use of frozen human skeletal muscle biopsies to simultaneously isolate and grow SCs and FAPs from healthy or dystrophic patients. We show that while the purified SCs differentiate into mature myotubes, purified FAPs can differentiate into adipocytes or fibroblasts demonstrating their multipotency. We find that these FAPs can be immortalized and the immortalized FAPs (iFAPs) retain their multipotency. These approaches open the door for carrying out personalized analysis of patient FAPs and interactions with the SCs that lead to muscle loss.

Keywords:  Duchenne muscular dystrophy; fibroadipogenic progenitors cells; muscle explant muscle biopsies; satellite cells

DOI:  https://doi.org/10.1096/fj.202100588R
Commun Biol. 2021 Aug 20. 4(1): 994

A xanthene derivative, DS20060511, attenuates glucose intolerance by inducing skeletal muscle-specific GLUT4 translocation in mice.

Shinji Furuzono, Tetsuya Kubota, Junki Taura, Masahiro Konishi, Asuka Naito, Masato Tsutsui, Hiroshi Karasawa, Naoto Kubota, Takashi Kadowaki.

Reduced glucose uptake into the skeletal muscle is an important pathophysiological abnormality in type 2 diabetes, and is caused by impaired translocation of glucose transporter 4 (GLUT4) to the skeletal muscle cell surface. Here, we show a xanthene derivative, DS20060511, induces GLUT4 translocation to the skeletal muscle cell surface, thereby stimulating glucose uptake into the tissue. DS20060511 induced GLUT4 translocation and stimulated glucose uptake into differentiated L6-myotubes and into the skeletal muscles in mice. These effects were completely abolished in GLUT4 knockout mice. Induction of GLUT4 translocation by DS20060511 was independent of the insulin signaling pathways including IRS1-Akt-AS160 phosphorylation and IRS1-Rac1-actin polymerization, eNOS pathway, and AMPK pathway. Acute and chronic DS20060511 treatment attenuated the glucose intolerance in obese diabetic mice. Taken together, DS20060511 acts as a skeletal muscle-specific GLUT4 translocation enhancer to facilitate glucose uptake. Further studies of DS20060511 may pave the way for the development of novel antidiabetic medicines.

DOI: https://doi.org/10.1038/s42003-021-02491-6
Front Physiol. 2021 ;12 685531

A Physiology-Guided Classification of Active-Stress and Active-Strain Approaches for Continuum-Mechanical Modeling of Skeletal Muscle Tissue.

Thomas Klotz, Christian Bleiler, Oliver Röhrle.

  The well-established sliding filament and cross-bridge theory explain the major biophysical mechanism responsible for a skeletal muscle's active behavior on a cellular level. However, the biomechanical function of skeletal muscles on the tissue scale, which is caused by the complex interplay of muscle fibers and extracellular connective tissue, is much less understood. Mathematical models provide one possibility to investigate physiological hypotheses. Continuum-mechanical models have hereby proven themselves to be very suitable to study the biomechanical behavior of whole muscles or entire limbs. Existing continuum-mechanical skeletal muscle models use either an active-stress or an active-strain approach to phenomenologically describe the mechanical behavior of active contractions. While any macroscopic constitutive model can be judged by it's ability to accurately replicate experimental data, the evaluation of muscle-specific material descriptions is difficult as suitable data is, unfortunately, currently not available. Thus, the discussions become more philosophical rather than following rigid methodological criteria. Within this work, we provide a extensive discussion on the underlying modeling assumptions of both the active-stress and the active-strain approach in the context of existing hypotheses of skeletal muscle physiology. We conclude that the active-stress approach resolves an idealized tissue transmitting active stresses through an independent pathway. In contrast, the active-strain approach reflects an idealized tissue employing an indirect, coupled pathway for active stress transmission. Finally the physiological hypothesis that skeletal muscles exhibit redundant pathways of intramuscular stress transmission represents the basis for considering a mixed-active-stress-active-strain constitutive framework.

Keywords:  active strain; active stress; continuum mechanics; muscle modeling; skeletal muscle

DOI:  https://doi.org/10.3389/fphys.2021.685531
Front Physiol. 2021 ;12 689492

Molecular and Biomechanical Adaptations to Mechanical Stretch in Cultured Myotubes.

Dapeng Ren, Jing Song, Ran Liu, Xuemin Zeng, Xiao Yan, Qiang Zhang, Xiao Yuan.

  Myotubes are mature muscle cells that form the basic structural element of skeletal muscle. When stretching skeletal muscles, myotubes are subjected to passive tension as well. This lead to alterations in myotube cytophysiology, which could be related with muscular biomechanics. During the past decades, much progresses have been made in exploring biomechanical properties of myotubes in vitro. In this review, we integrated the studies focusing on cultured myotubes being mechanically stretched, and classified these studies into several categories: amino acid and glucose uptake, protein turnover, myotube hypertrophy and atrophy, maturation, alignment, secretion of cytokines, cytoskeleton adaption, myotube damage, ion channel activation, and oxidative stress in myotubes. These biomechanical adaptions do not occur independently, but interconnect with each other as part of the systematic mechanoresponse of myotubes. The purpose of this review is to broaden our comprehensions of stretch-induced muscular alterations in cellular and molecular scales, and to point out future challenges and directions in investigating myotube biomechanical manifestations.

Keywords:  biomechanical adaptation; mechanical stretch; mechanoresponse; myotube; skeletal muscle

DOI:  https://doi.org/10.3389/fphys.2021.689492
Appl Physiol Nutr Metab. 2021 Aug 16.

Effect of Local Heat Application During Exercise on Gene Expression Related to Mitochondrial Homeostasis.

Nattie O'Reilly, Christopher Collins, Mark L McGlynn, Dustin Russel Slivka.

Determine the impact of local muscle heating during endurance exercise on human skeletal muscle mitochondrial-related gene expression. Twelve subjects (25±6 yrs, 177±8 cm, 78±16 kg, and VO2peak peak 45±8 ml·kg-1·min-1) cycled with one leg heated (HOT) and the other serving as a control (CON). Skin and intramuscular temperatures were taken before temperature intervention (Pre), after 30 min (Pre30), after exercise (Post) and four hours after exercise (4Post). Muscle biopsies were taken from each leg at Pre and 4Post. Intramuscular temperature increased within HOT (34.4±0.7ºC to 36.1±0.5ºC, p<0.001) and was higher than CON at Pre30 (34.0±0.7ºC, p<0.001). However, temperatures at POST were similar (HOT 38.4±0.7ºC, CON 38.3±0.5ºC, p=0.661). Skin temperature was higher than CON at Post30 (30.3±1.0ºC, p<0.001) and Post (HOT 34.6±0.9ºC, CON 32.3±1.6ºC, p<0.001). PGC-1α, VEGF and NRF2 mRNA increased with exercise (p<0.05) but was not altered with heating (p>0.05). TFAM increased after exercise with heat application (HOT, p=0.019) but not with exercise alone (CON, p=0.422). There was no difference in NRF1, ESRRα, or any of the mitophagy related genes in response to exercise or temperature (p>0.05). In conclusion, TFAM is enhanced by local heat application during endurance exercise, whereas other genes related to mitochondrial homeostasis are unaffected. Novelty: The main finding of this study is that localized heating increased TFAM mRNA expression. The normal exercise-induced increased PGC-1α gene expression was unaltered by local muscle heating.

DOI: https://doi.org/10.1139/apnm-2021-0346
Transcription. 2021 Aug 17. 1-17

CREB-mediated transcriptional activation of NRMT1 drives muscle differentiation.

John G Tooley, James P Catlin, Christine E Schaner Tooley.

  The N-terminal methyltransferase NRMT1 is an important regulator of protein/DNA interactions and plays a role in many cellular processes, including mitosis, cell cycle progression, chromatin organization, DNA damage repair, and transcriptional regulation. Accordingly, loss of NRMT1 results in both developmental pathologies and oncogenic phenotypes. Though NRMT1 plays such important and diverse roles in the cell, little is known about its own regulation. To better understand the mechanisms governing NRMT1 expression, we first identified its predominant transcriptional start site and minimal promoter region with predicted transcription factor motifs. We then used a combination of luciferase and binding assays to confirm CREB1 as the major regulator of NRMT1 transcription. We tested which conditions known to activate CREB1 also activated NRMT1 transcription, and found CREB1-mediated NRMT1 expression was increased during recovery from serum starvation and muscle cell differentiation. To determine how NRMT1 expression affects myoblast differentiation, we used CRISPR/Cas9 technology to knock out NRMT1 expression in immortalized C2C12 mouse myoblasts. C2C12 cells depleted of NRMT1 lacked Pax7 expression and were unable to proceed down the muscle differentiation pathway. Instead, they took on characteristics of C2C12 cells that have transdifferentiated into osteoblasts, including increased alkaline phosphatase and type I collagen expression and decreased proliferation. These data implicate NRMT1 as an important downstream target of CREB1 during muscle cell differentiation.

Keywords:  CREB1; NRMT1; differentiation; methylation; stem cell

DOI:  https://doi.org/10.1080/21541264.2021.1963627
Hum Mol Genet. 2021 Aug 14. pii: ddab230. [Epub ahead of print]

Myeloid cell-mediated targeting of LIF to dystrophic muscle causes transient increases in muscle fiber lesions by disrupting the recruitment and dispersion of macrophages in muscle.

Ivan Flores, Steven S Welc, Michelle Wehling-Henricks, James G Tidball.

Leukemia inhibitory factor (LIF) can influence development by increasing cell proliferation and inhibiting differentiation. Because of its potency for expanding stem cell populations, delivery of exogenous LIF to diseased tissue could have therapeutic value. However, systemic elevations of LIF can have negative, off-target effects. We tested whether inflammatory cells expressing a LIF transgene under control of a leukocyte-specific, CD11b promoter provide a strategy to target LIF to sites of damage in the mdx mouse model of Duchenne muscular dystrophy, leading to increased numbers of muscle stem cells and improved muscle regeneration. However, transgene expression in inflammatory cells did not increase muscle growth or increase numbers of stem cells required for regeneration. Instead, transgene expression disrupted the normal dispersion of macrophages in dystrophic muscles, leading to transient increases in muscle damage in foci where macrophages were highly-concentrated during early stages of pathology. The defect in inflammatory cell dispersion reflected impaired chemotaxis of macrophages to C-C motif chemokine ligand-2 and local increases of LIF production that produced large aggregations of cytolytic macrophages. Transgene expression also induced a shift in macrophage phenotype away from a CD206+, M2-biased phenotype that supports regeneration. However, at later stages of the disease when macrophage numbers declined, they dispersed in the muscle, leading to reductions in muscle fiber damage, compared to non-transgenic mdx mice. Together, the findings show that macrophage-mediated delivery of transgenic LIF exerts differential effects on macrophage dispersion and muscle damage depending on the stage of dystrophic pathology.

DOI: https://doi.org/10.1093/hmg/ddab230
Front Mol Biosci. 2021 ;8 685993

Cold-Induced RNA-Binding Protein Promotes Glucose Metabolism and Reduces Apoptosis by Increasing AKT Phosphorylation in Mouse Skeletal Muscle Under Acute Cold Exposure.

Yang Liu, Peng Liu, Yajie Hu, Yu Cao, Jingjing Lu, Yuying Yang, Hongming Lv, Shuai Lian, Bin Xu, Shize Li.

  The main danger of cold stress to animals in cold regions is systemic metabolic changes and protein synthesis inhibition. Cold-induced RNA-binding protein is a cold shock protein that is rapidly up-regulated under cold stimulation in contrast to the inhibition of most proteins and participates in multiple cellular physiological activities by regulating targets. Therefore, this study was carried out to investigate the possible mechanism of CIRP-mediated glucose metabolism regulation and survival promotion in skeletal muscle after acute cold exposure. Skeletal muscle and serum from mice were obtained after 0, 2, 4 and 8 h of acute hypothermia exposure. Subsequently, the changes of CIRP, metabolism and apoptosis were examined. Acute cold exposure increased energy consumption, enhanced glycolysis, increased apoptosis, and up-regulated CIRP and phosphorylation of AKT. In addition, CIRP overexpression in C2C12 mouse myoblasts at each time point under 37°C and 32°C mild hypothermia increased AKT phosphorylation, enhanced glucose metabolism, and reduced apoptosis. CIRP knockdown by siRNA interference significantly reduced the AKT phosphorylation of C2C12 cells. Wortmannin inhibited the AKT phosphorylation of skeletal muscle after acute cold exposure, thereby inhibiting glucose metabolism and aggravating apoptosis. Taken together, acute cold exposure up-regulates CIRP in mouse skeletal muscle, which regulates glucose metabolism and maintains energy balance in skeletal muscle cells through the AKT signaling pathway, thus slowing down the apoptosis of skeletal muscle cells.

Keywords:  Akt; CIRP; acute cold exposure; apoptosis; glucose metabolism

DOI:  https://doi.org/10.3389/fmolb.2021.685993
Mol Ther. 2021 Aug 13. pii: S1525-0016(21)00414-7. [Epub ahead of print]

Cell-type selective targeted delivery of a recombinant lysosomal enzyme for enzyme therapies.

Andrew D Baik, Philip Calafati, Xiaoli Zhang, Nina A Aaron, Antonia Mehra, Sven Moller-Tank, Lawrence Miloscio, Maria Praggastis, Nicholas Giovannone, Cheryl Pan, Yajun Tang, Susannah Bridges, Alejo Mujica, Peter Barbounis, Jean Yanolatos, Nicholas Gale, Ning Li, Christos A Kyratsous, Christopher J Schoenherr, Andrew J Murphy, Aris N Economides, Katherine D Cygnar.

Lysosomal diseases are a class of genetic disorders predominantly caused by loss of lysosomal hydrolases, leading to lysosomal and cellular dysfunction. Enzyme replacement therapy (ERT), where recombinant enzyme is given intravenously, internalized by cells, and trafficked to the lysosome, has been applied to treat several lysosomal diseases. However, current ERT regimens do not correct disease phenotypes in all affected organs because the biodistribution of enzyme uptake does not match that of the affected cells that require the enzyme. We present here targeted ERT, an approach that utilizes antibody-enzyme fusion proteins to target the enzyme to specific cell types. The antibody moiety recognizes transmembrane proteins involved in lysosomal trafficking and that are also preferentially expressed in those cells most affected in disease. Using Pompe disease (PD) as an example, we show that targeted ERT is superior to ERT in treating the skeletal muscle phenotypes of PD mice both as a protein replacement therapeutic and as a gene therapy.

DOI: https://doi.org/10.1016/j.ymthe.2021.08.020
Nat Commun. 2021 08 19. 12(1): 5043

Translational control by DHX36 binding to 5'UTR G-quadruplex is essential for muscle stem-cell regenerative functions.

Xiaona Chen, Jie Yuan, Guang Xue, Silvia Campanario, Di Wang, Wen Wang, Xi Mou, Shiau Wei Liew, Mubarak Ishaq Umar, Joan Isern, Yu Zhao, Liangqiang He, Yuying Li, Christopher J Mann, Xiaohua Yu, Lei Wang, Eusebio Perdiguero, Wei Chen, Yuanchao Xue, Yoshikuni Nagamine, Chun Kit Kwok, Hao Sun, Pura Muñoz-Cánoves, Huating Wang.

Skeletal muscle has a remarkable ability to regenerate owing to its resident stem cells (also called satellite cells, SCs). SCs are normally quiescent; when stimulated by damage, they activate and expand to form new fibers. The mechanisms underlying SC proliferative progression remain poorly understood. Here we show that DHX36, a helicase that unwinds RNA G-quadruplex (rG4) structures, is essential for muscle regeneration by regulating SC expansion. DHX36 (initially named RHAU) is barely expressed at quiescence but is highly induced during SC activation and proliferation. Inducible deletion of Dhx36 in adult SCs causes defective proliferation and muscle regeneration after damage. System-wide mapping in proliferating SCs reveals DHX36 binding predominantly to rG4 structures at various regions of mRNAs, while integrated polysome profiling shows that DHX36 promotes mRNA translation via 5'-untranslated region (UTR) rG4 binding. Furthermore, we demonstrate that DHX36 specifically regulates the translation of Gnai2 mRNA by unwinding its 5' UTR rG4 structures and identify GNAI2 as a downstream effector of DHX36 for SC expansion. Altogether, our findings uncover DHX36 as an indispensable post-transcriptional regulator of SC function and muscle regeneration acting through binding and unwinding rG4 structures at 5' UTR of target mRNAs.

DOI: https://doi.org/10.1038/s41467-021-25170-w
Diabetes Metab Res Rev. 2021 Aug 16. e3490

Branched-chain amino acids at supraphysiological but not physiological levels reduce myotube insulin sensitivity.

Madison E Rivera, Caroline N Rivera, Roger A Vaughan.

   AIMS: Branched-chain amino acids (BCAA) are often emphasized in the diets of avid exercisers, yet population data demonstrates a correlation between circulating BCAA and insulin resistance. However, it is unclear if BCAA independently promote insulin resistance in otherwise healthy cells. The purpose of this study is to examine the effect of a BCAA mixture on muscle insulin signaling in vitro in both insulin resistant and sensitive cells.
MATERIALS AND METHODS: C2C12 myotubes were treated with a BCAA mixture containing leucine:isoleucine:valine at a ratio of 2:1:1 at 0.2mM, 2mM, or 20mM (based on leucine content) for either 30 min, 1 day, or 6 days. Western blot was used to assess insulin sensitivity of cells treated with BCAA both with and without concurrent insulin resistance, and, with and without insulin stimulation.
RESULTS: BCAA treatment for 1 day significantly reduced basal, but not insulin-stimulated pAkt expression. BCAA treatment for 6 days resulted in significantly reduced basal insulin signaling in health cells and insulin-stimulated insulin signaling in insulin resistant (but not insulin sensitive) cells.
CONCLUSION: Similar to previous observations demonstrating BCAA may correlate with insulin resistance during metabolically stressed conditions, we demonstrate excessively high BCAA exposure can negatively influence basal insulin signaling, as well as insulin sensitivity in insulin resistant myotubes. However, given the intentionally high concentrations of BCAA used in this study, the extent to which these observations translate to in vivo models is unclear and warrants further investigation. This article is protected by copyright. All rights reserved.

Keywords:  Leucine; diabetes; insulin resistance; isoleucine; pAkt/Akt; skeletal muscle; valine

DOI:  https://doi.org/10.1002/dmrr.3490
J Physiol. 2021 Aug 15.

Extracellular miRNAs as mediators of obesity-associated disease.

Bruna Brasil Brandao, Marsel Lino, C Ronald Kahn.

  Extracellular miRNAs are found in a variety of body fluids and mediate intercellular and interorgan communication thus regulating gene expression and cellular metabolism. These miRNAs are secreted either in small vesicles/exosomes (sEV) or bound to proteins such as Argonaute and HDL. Both exosomal and protein-bound circulating miRNAs are altered in obesity. Although all tissues can contribute to changes in circulating miRNAs, adipose tissue itself is an important source of these miRNAs, especially those in sEVs. These are derived from both adipocytes and macrophages and participate in crosstalk between these cells, as well as peripheral tissues, including liver, skeletal muscle, and pancreas, whose function may be impaired in obesity. Changes in levels of circulating miRNAs have also been linked to the beneficial effects induced by weight loss interventions, including diet, exercise, and bariatric surgery, further indicating a role for these miRNAs as mediators of disease pathogenesis. Here, we review the role of circulating miRNAs in the pathophysiology of obesity and explore their potential use as biomarkers and in therapy of obesity-associated metabolic syndrome. Abstract figure legend Extracellular miRNAs as indicators of metabolic status. Extracellular miRNAs are found in the circulation and are secreted bound to protein complex, lipoproteins particles or loaded into extracellular vesicles including microvesicles and exosomes. Many circulating microRNAs are dysregulated during obesity, but weight loss intervention such as exercise, diet and bariatric surgery can restore the miRNA secretion. Metabolic organs including adipose tissue, liver, skeletal muscle, and pancreas, whose function may be impaired during obesity, contribute to changes in the pool of obesity-associated circulating miRNAs. This article is protected by copyright. All rights reserved.

Keywords:  RNA therapy; biomarkers; circulating miRNAs; exosomes; obesity; tissue-crosstalk

DOI:  https://doi.org/10.1113/JP280910
FASEB J. 2021 Sep;35(9): e21860

A mutation in desmin makes skeletal muscle less vulnerable to acute muscle damage after eccentric loading in rats.

Henning T Langer, Agata A Mossakowski, Alec M Avey, Ross P Wohlgemuth, Lucas R Smith, Herman Zbinden-Foncea, Keith Baar.

  Desminopathy is the most common intermediate filament disease in humans. The most frequent mutation causing desminopathy in patients is a R350P DES missense mutation. We have developed a rat model with an analogous mutation in R349P Des. To investigate the role of R349P Des in mechanical loading, we stimulated the sciatic nerve of wild-type littermates (WT) (n = 6) and animals carrying the mutation (MUT) (n = 6) causing a lengthening contraction of the dorsi flexor muscles. MUT animals showed signs of ongoing regeneration at baseline as indicated by a higher number of central nuclei (genotype: P < .0001). While stimulation did not impact central nuclei, we found an increased number of IgG positive fibers (membrane damage indicator) after eccentric contractions with both genotypes (stimulation: P < .01). Interestingly, WT animals displayed a more pronounced increase in IgG positive fibers with stimulation compared to MUT (interaction: P < .05). In addition to altered histology, molecular signaling on the protein level differed between WT and MUT. The membrane repair protein dysferlin decreased with eccentric loading in WT but increased in MUT (interaction: P < .05). The autophagic substrate p62 was increased in both genotypes with loading (stimulation: P < .05) but tended to be more elevated in WT (interaction: P = .05). Caspase 3 levels, a central regulator of apoptotic cell death, was increased with stimulation in both genotypes (stimulation: P < .01) but more so in WT animals (interaction: P < .0001). Overall, our data indicate that R349P Des rats have a lower susceptibility to structural muscle damage of the cytoskeleton and sarcolemma with acute eccentric loading.

Keywords:  exercise; filament; injury; intermediate; muscle; signaling

DOI:  https://doi.org/10.1096/fj.202100711RR
Genes Dev. 2021 Aug 19.

Dissecting dual roles of MyoD during lineage conversion to mature myocytes and myogenic stem cells.

Masaki Yagi, Fei Ji, Jocelyn Charlton, Simona Cristea, Kathleen Messemer, Naftali Horwitz, Bruno Di Stefano, Nikolaos Tsopoulidis, Michael S Hoetker, Aaron J Huebner, Ori Bar-Nur, Albert E Almada, Masakazu Yamamoto, Anthony Patelunas, David J Goldhamer, Amy J Wagers, Franziska Michor, Alexander Meissner, Ruslan I Sadreyev, Konrad Hochedlinger.

  The generation of myotubes from fibroblasts upon forced MyoD expression is a classic example of transcription factor-induced reprogramming. We recently discovered that additional modulation of signaling pathways with small molecules facilitates reprogramming to more primitive induced myogenic progenitor cells (iMPCs). Here, we dissected the transcriptional and epigenetic dynamics of mouse fibroblasts undergoing reprogramming to either myotubes or iMPCs using a MyoD-inducible transgenic model. Induction of MyoD in fibroblasts combined with small molecules generated Pax7+ iMPCs with high similarity to primary muscle stem cells. Analysis of intermediate stages of iMPC induction revealed that extinction of the fibroblast program preceded induction of the stem cell program. Moreover, key stem cell genes gained chromatin accessibility prior to their transcriptional activation, and these regions exhibited a marked loss of DNA methylation dependent on the Tet enzymes. In contrast, myotube generation was associated with few methylation changes, incomplete and unstable reprogramming, and an insensitivity to Tet depletion. Finally, we showed that MyoD's ability to bind to unique bHLH targets was crucial for generating iMPCs but dispensable for generating myotubes. Collectively, our analyses elucidate the role of MyoD in myogenic reprogramming and derive general principles by which transcription factors and signaling pathways cooperate to rewire cell identity.

Keywords:  DNA methylation; MyoD; dedifferentiation; epigenetic reprogramming; induced myogenic progenitor cells (iMPCs); satellite cells; transdifferentiation

DOI:  https://doi.org/10.1101/gad.348678.121
Diabetologia. 2021 Aug 14.

Normal to enhanced intrinsic mitochondrial respiration in skeletal muscle of middle- to older-aged women and men with uncomplicated type 1 diabetes.

Cynthia M F Monaco, Mark A Tarnopolsky, Athan G Dial, Joshua P Nederveen, Irena A Rebalka, Maria Nguyen, Lauren V Turner, Christopher G R Perry, Vladimir Ljubicic, Thomas J Hawke.

   AIMS/HYPOTHESIS: This study interrogated mitochondrial respiratory function and content in skeletal muscle biopsies of healthy adults between 30 and 72 years old with and without uncomplicated type 1 diabetes.
METHODS: Participants (12 women/nine men) with type 1 diabetes (48 ± 11 years of age), without overt complications, were matched for age, sex, BMI and level of physical activity to participants without diabetes (control participants) (49 ± 12 years of age). Participants underwent a Bergström biopsy of the vastus lateralis to assess mitochondrial respiratory function using high-resolution respirometry and citrate synthase activity. Electron microscopy was used to quantify mitochondrial content and cristae (pixel) density.
RESULTS: Mean mitochondrial area density was 27% lower (p = 0.006) in participants with type 1 diabetes compared with control participants. This was largely due to smaller mitochondrial fragments in women with type 1 diabetes (-18%, p = 0.057), as opposed to a decrease in the total number of mitochondrial fragments in men with diabetes (-28%, p = 0.130). Mitochondrial respiratory measures, whether estimated per milligram of tissue (i.e. mass-specific) or normalised to area density (i.e. intrinsic mitochondrial function), differed between cohorts, and demonstrated sexual dimorphism. Mass-specific mitochondrial oxidative phosphorylation (OXPHOS) capacity with the substrates for complex I and complex II (CI + II) was significantly lower (-24%, p = 0.033) in women with type 1 diabetes compared with control participants, whereas mass-specific OXPHOS capacities with substrates for complex I only (pyruvate [CI pyr] or glutamate [CI glu]) or complex II only (succinate [CII succ]) were not different (p > 0.404). No statistical differences (p > 0.397) were found in mass-specific OXPHOS capacity in men with type 1 diabetes compared with control participants despite a 42% non-significant increase in CI glu OXPHOS capacity (p = 0.218). In contrast, intrinsic CI + II OXPHOS capacity was not different in women with type 1 diabetes (+5%, p = 0.378), whereas in men with type 1 diabetes it was 25% higher (p = 0.163) compared with control participants. Men with type 1 diabetes also demonstrated higher intrinsic OXPHOS capacity for CI pyr (+50%, p = 0.159), CI glu (+88%, p = 0.033) and CII succ (+28%, p = 0.123), as well as higher intrinsic respiratory rates with low (more physiological) concentrations of either ADP, pyruvate, glutamate or succinate (p < 0.012). Women with type 1 diabetes had higher (p < 0.003) intrinsic respiratory rates with low concentrations of succinate only. Calculated aerobic fitness (Physical Working Capacity Test [PWC130]) showed a strong relationship with mitochondrial respiratory function and content in the type 1 diabetes cohort.
CONCLUSIONS/INTERPRETATION: In middle- to older-aged adults with uncomplicated type 1 diabetes, we conclude that skeletal muscle mitochondria differentially adapt to type 1 diabetes and demonstrate sexual dimorphism. Importantly, these cellular alterations were significantly associated with our metric of aerobic fitness (PWC130) and preceded notable impairments in skeletal mass and strength.

Keywords:  Aerobic fitness; Mitochondria; Older adults; Oxidative phosphorylation; Skeletal muscle; Type 1 diabetes

DOI:  https://doi.org/10.1007/s00125-021-05540-1
Cell Physiol Biochem. 2021 Aug 20. 55(4): 489-504

Nox4 Knockout Does Not Prevent Diaphragm Atrophy, Contractile Dysfunction, or Mitochondrial Maladaptation in the Early Phase Post-Myocardial Infarction in Mice.

Dongwoo Hahn, Ravi A Kumar, Derek R Muscato, Terence E Ryan, Katrin Schröder, Leonardo F Ferreira.

   BACKGROUND/AIMS: Diaphragm dysfunction with increased reactive oxygen species (ROS) occurs within 72 hrs post-myocardial infarction (MI) in mice and may contribute to loss of inspiratory maximal pressure and endurance in patients.
METHODS: We used wild-type (WT) and whole-body Nox4 knockout (Nox4KO) mice to measure diaphragm bundle force in vitro with a force transducer, mitochondrial respiration in isolated fiber bundles with an O2 sensor, mitochondrial ROS by fluorescence, mRNA (RT-PCR) and protein (immunoblot), and fiber size by histology 72 hrs post-MI.
RESULTS: MI decreased diaphragm fiber cross-sectional area (CSA) (~15%, p = 0.015) and maximal specific force (10%, p = 0.005), and increased actin carbonylation (5-10%, p = 0.007) in both WT and Nox4KO. Interestingly, MI did not affect diaphragm mRNA abundance of MAFbx/atrogin-1 and MuRF-1 but Nox4KO decreased it by 20-50% (p < 0.01). Regarding the mitochondria, MI and Nox4KO decreased the protein abundance of citrate synthase and subunits of electron transport system (ETS) complexes and increased mitochondrial O2 flux (JO2) and H2O2 emission (JH2O2) normalized to citrate synthase. Mitochondrial electron leak (JH2O2/JO2) in the presence of ADP was lower in Nox4KO and not changed by MI.
CONCLUSION: Our study shows that the early phase post-MI causes diaphragm atrophy, contractile dysfunction, sarcomeric actin oxidation, and decreases citrate synthase and subunits of mitochondrial ETS complexes. These factors are potential causes of loss of inspiratory muscle strength and endurance in patients, which likely contribute to the pathophysiology in the early phase post-MI. Whole-body Nox4KO did not prevent the diaphragm abnormalities induced 72 hrs post-MI, suggesting that systemic pharmacological inhibition of Nox4 will not benefit patients in the early phase post-MI.

Keywords:  Atrophy; Oxidants; Force; Respiration; Heart failure

DOI:  https://doi.org/10.33594/000000400
Circ Res. 2021 Aug 18.

Filamin C Cardiomyopathy Variants Cause Protein and Lysosome Accumulation.

Radhika Agarwal, Joao Paulo, Christopher N Toepfer, Jourdan K Ewoldt, Subramanian Sundaram, Anant Chopra, Qi Zhang, Joshua Gorham, Steven DePalma, Christopher S Chen, Steven Gygi, Christine E Seidman, Jonathan G Seidman.

Rationale: Dominant heterozygous variants in Filamin C (FLNC) cause diverse cardiomyopathies, though the underlying molecular mechanisms remain poorly understood. Objective: We aimed to define the molecular mechanisms by which FLNC variants altered human cardiomyocyte gene and protein expression, sarcomere structure, and contractile performance. Methods and Results: Using CRISPR/Cas9, we introduced FLNC variants into human cardiomyocytes derived from induced pluripotent stem cells (hiPSC-CMs). We compared isogenic hiPSC-CMs with normal (WT), ablated expression (FLNC-/-) or haploinsufficiency (FLNC+/-) that causes DCM. We also studied a heterozygous in-frame deletion (FLNC+/∆7aa) which did not affect FLNC expression but caused aggregate formation, similar to FLNC variants associated with hypertrophic cardiomyopathy (HCM). FLNC-/- hiPSC-CMs demonstrated profound sarcomere misassembly and reduced contractility. While sarcomere formation and function were unaffected in FLNC+/- and FLNC+/∆7aa hiPSC-CMs, these heterozygous variants caused increases in lysosome content, enhancement of autophagic flux, and accumulation of FLNC-binding partners and Z-disc proteins. Conclusions: FLNC expression is required for sarcomere organization and physiologic function. Variants that produce misfolded FLNC proteins cause the accumulation of FLNC and FLNC binding partners which leads to increased lysosome expression and activation of autophagic pathways. Surprisingly, similar pathways were activated in FLNC haploinsufficient hiPSC-CMs, likely initiated by the loss of stoichiometric FLNC protein interactions and impaired turnover of proteins at the Z-disc. These results indicate that both FLNC haploinsufficient variants and variants that produce misfolded FLNC protein cause disease by similar proteotoxic mechanisms, and indicate the therapeutic potential for augmenting protein degradative pathways to treat a wide range of FLNC-related cardiomyopathies.

DOI: https://doi.org/10.1161/CIRCRESAHA.120.317076
Front Genet. 2021 ;12 702547

Myogenic Cell Transplantation in Genetic and Acquired Diseases of Skeletal Muscle.

Study Group

  This article will review myogenic cell transplantation for congenital and acquired diseases of skeletal muscle. There are already a number of excellent reviews on this topic, but they are mostly focused on a specific disease, muscular dystrophies and in particular Duchenne Muscular Dystrophy. There are also recent reviews on cell transplantation for inflammatory myopathies, volumetric muscle loss (VML) (this usually with biomaterials), sarcopenia and sphincter incontinence, mainly urinary but also fecal. We believe it would be useful at this stage, to compare the same strategy as adopted in all these different diseases, in order to outline similarities and differences in cell source, pre-clinical models, administration route, and outcome measures. This in turn may help to understand which common or disease-specific problems have so far limited clinical success of cell transplantation in this area, especially when compared to other fields, such as epithelial cell transplantation. We also hope that this may be useful to people outside the field to get a comprehensive view in a single review. As for any cell transplantation procedure, the choice between autologous and heterologous cells is dictated by a number of criteria, such as cell availability, possibility of in vitro expansion to reach the number required, need for genetic correction for many but not necessarily all muscular dystrophies, and immune reaction, mainly to a heterologous, even if HLA-matched cells and, to a minor extent, to the therapeutic gene product, a possible antigen for the patient. Finally, induced pluripotent stem cell derivatives, that have entered clinical experimentation for other diseases, may in the future offer a bank of immune-privileged cells, available for all patients and after a genetic correction for muscular dystrophies and other myopathies.

Keywords:  cell transplantation; inflammatory myopathies; mitochondrial myopathies; muscle stem cells; muscular dystrophies; sphincter incontinence; volumetric muscle loss

DOI:  https://doi.org/10.3389/fgene.2021.702547
Physiol Rep. 2021 Aug;9(16): e14961

Muscle-specific sirtuin 3 overexpression does not attenuate the pathological effects of high-fat/high-sucrose feeding but does enhance cardiac SERCA2a activity.

Christopher J Oldfield, Teri L Moffatt, Kimberley A O'Hara, Bo Xiang, Vernon W Dolinsky, Todd A Duhamel.

  Obesity, type 2 diabetes, and heart disease are linked to an unhealthy diet. Sarco(endo)plasmic reticulum calcium (Ca2+ ) ATPase 2a (SERCA2a) controls cardiac function by transporting Ca2+ in cardiomyocytes. SERCA2a is altered by diet and acetylation, independently; however, it is unknown if diet alters cardiac SERCA2a acetylation. Sirtuin (SIRT) 3 is an enzyme that might preserve health under conditions of macronutrient excess by modulating metabolism via regulating deacetylation of target proteins. Our objectives were to determine if muscle-specific SIRT3 overexpression attenuates the pathological effects of high fat-high sucrose (HFHS) feeding and if HFHS feeding alters cardiac SERCA2a acetylation. We also determined if SIRT3 alters cardiac SERCA2a acetylation and regulates cardiac SERCA2a activity. C57BL/6J wild-type (WT) mice and MCK-mSIRT3-M1-Flag transgenic (SIRT3TG ) mice, overexpressing SIRT3 in cardiac and skeletal muscle, were fed a standard-diet or a HFHS-diet for 4 months. SIRT3TG and WT mice developed obesity, glucose intolerance, cardiac dysfunction, and pathological cardiac remodeling after 4 months of HFHS feeding, indicating muscle-specific SIRT3 overexpression does not attenuate the pathological effects of HFHS-feeding. Overall cardiac lysine acetylation was increased by 63% in HFHS-fed mice (p = 0.022), though HFHS feeding did not alter cardiac SERCA2a acetylation. Cardiac SERCA2a acetylation was not altered by SIRT3 overexpression, whereas SERCA2a Vmax was 21% higher in SIRT3TG (p = 0.039) than WT mice. This suggests that SIRT3 overexpression enhanced cardiac SERCA2a activity without direct SERCA2a deacetylation. Muscle-specific SIRT3 overexpression may not prevent the complications associated with an unhealthy diet in mice, but it appears to enhance SERCA2a activity in the mouse heart.

Keywords:  SERCA; acetylation; calcium handling; diabetes; obesity; sirtuins

DOI:  https://doi.org/10.14814/phy2.14961
BMC Sports Sci Med Rehabil. 2021 Aug 14. 13(1): 90

The effects of cocoa flavanols on indices of muscle recovery and exercise performance: a narrative review.

Liam D Corr, Adam Field, Deborah Pufal, Tom Clifford, Liam D Harper, Robert J Naughton.

  Exercise-induced muscle damage (EIMD) is associated with oxidative stress and inflammation, muscle soreness, and reductions in muscle function. Cocoa flavanols (CF) are (poly)phenols with antioxidant and anti-inflammatory effects and thus may attenuate symptoms of EIMD. The purpose of this narrative review was to collate and evaluate the current literature investigating the effect of CF supplementation on markers of exercise-induced oxidative stress and inflammation, as well as changes in muscle function, perceived soreness, and exercise performance. Acute and sub-chronic intake of CF reduces oxidative stress resulting from exercise. Evidence for the effect of CF on exercise-induced inflammation is lacking and the impact on muscle function, perceived soreness and exercise performance is inconsistent across studies. Supplementation of CF may reduce exercise-induced oxidative stress, with potential for delaying fatigue, but more evidence is required for any definitive conclusions on the impact of CF on markers of EIMD.

Keywords:  Dark chocolate; Fatigue; Inflammation; Muscle damage; Muscle function; Oxidative stress

DOI:  https://doi.org/10.1186/s13102-021-00319-8
EMBO Rep. 2021 Aug 17. e48018

Titin kinase ubiquitination aligns autophagy receptors with mechanical signals in the sarcomere.

Julius Bogomolovas, Jennifer R Fleming, Barbara Franke, Bruno Manso, Bernd Simon, Alexander Gasch, Marija Markovic, Thomas Brunner, Ralph Knöll, Ju Chen, Siegfried Labeit, Martin Scheffner, Christine Peter, Olga Mayans.

  Striated muscle undergoes remodelling in response to mechanical and physiological stress, but little is known about the integration of such varied signals in the myofibril. The interaction of the elastic kinase region from sarcomeric titin (A168-M1) with the autophagy receptors Nbr1/p62 and MuRF E3 ubiquitin ligases is well suited to link mechanosensing with the trophic response of the myofibril. To investigate the mechanisms of signal cross-talk at this titin node, we elucidated its 3D structure, analysed its response to stretch using steered molecular dynamics simulations and explored its functional relation to MuRF1 and Nbr1/p62 using cellular assays. We found that MuRF1-mediated ubiquitination of titin kinase promotes its scaffolding of Nbr1/p62 and that the process can be dynamically down-regulated by the mechanical unfolding of a linker sequence joining titin kinase with the MuRF1 receptor site in titin. We propose that titin ubiquitination is sensitive to the mechanical state of the sarcomere, the regulation of sarcomere targeting by Nbr1/p62 being a functional outcome. We conclude that MuRF1/Titin Kinase/Nbr1/p62 constitutes a distinct assembly that predictably promotes sarcomere breakdown in inactive muscle.

Keywords:  X-ray crystallography; cellular signalling; mechanotransduction; steered molecular dynamics simulations; ubiquitination

DOI:  https://doi.org/10.15252/embr.201948018