bims-moremu 2023-09-24 papers

bims-moremu

Biomed News

on Molecular regulators of muscle mass

Issue of 2023–09–24
24 papers selected by
Anna Vainshtein, Craft Science Inc.

Semaphorins: Missing Signals in Age-dependent Alteration of Neuromuscular Junctions and Skeletal Muscle Regeneration.
The impact and utility of very low-calorie diets: the role of exercise and protein in preserving skeletal muscle mass.
Absolute Quantification of mRNA Isoforms in Adult Stem Cells Using Microfluidic Digital PCR.
Advances in research on cell models for skeletal muscle atrophy.
Mimicking sarcolemmal damagein vitro: a contractile 3D model of skeletal muscle for drug testing in Duchenne muscular dystrophy.
Irisin ameliorated skeletal muscle atrophy by inhibiting fatty acid oxidation and pyroptosis induced by palmitic acid in chronic kidney disease.
Regulatory T cells require IL6 receptor alpha signaling to control skeletal muscle function and regeneration.
Stimulator of interferon genes promotes diabetic sarcopenia by targeting peroxisome proliferator activated receptors γ degradation and inhibiting fatty acid oxidation.
Migration and differentiation of muscle stem cells are coupled by RhoA signalling during regeneration.
Both moderate- and high-intensity exercise training increase intramyocellular lipid droplet abundance and modify myocellular distribution in adults with obesity.
Loss of Trim28 in muscle alters mitochondrial signalling but not systemic metabolism.
Diosgenin prevents dexamethasone-induced myotube atrophy in C2C12 cells.
Exhaustive exercise alters native and site-specific H2O2 emission in red and white skeletal muscle mitochondria.
KLF4 regulates skeletal muscle development and regeneration by directly targeting P57 and Myomixer.
Muscle: an independent contributor to the neuromuscular spinal muscular atrophy disease phenotype.
Acyl-CoA synthetase expression in human skeletal muscle is reduced in obesity and insulin resistance.
The role of estrogen in female skeletal muscle aging: A systematic review.
Stimulation of fat oxidation in rat muscle by unacylated ghrelin persists for 2-3 hours, but is independent of fatty acid transporter translocation.
Highlighting the idea of exerkines in the management of cancer patients with cachexia: novel insights and a critical review.
Modelling three-dimensional cancer-associated cachexia and therapy: The molecular basis and therapeutic potential of interleukin-6 transignalling blockade.
Iditarod, a Drosophila homolog of the Irisin precursor FNDC5, is critical for exercise performance and cardiac autophagy.
The separation and identification of circulating small extracellular vesicles from endurance-trained, strength-trained and recreationally active men.
Influence of Inflammatory Cytokines IL-1β and IFNγ on Sarcoplasmic Aggregation of p62 and TDP-43 in Myotubes.
Effects of fatigue on intra-muscle force-stabilizing synergies.

Aging Dis. 2023 Sep 03.

Semaphorins: Missing Signals in Age-dependent Alteration of Neuromuscular Junctions and Skeletal Muscle Regeneration.

Damon Fard, Alessandra Barbiera, Gabriella Dobrowolny, Luca Tamagnone, Bianca Maria Scicchitano.

Skeletal muscle is characterized by a remarkable capacity to rearrange after physiological changes and efficiently regenerate. However, during aging, extensive injury, or pathological conditions, the complete regenerative program is severely affected, with a progressive loss of muscle mass and function, a condition known as sarcopenia. The compromised tissue repair program is attributable to the gradual depletion of stem cells and to altered regulatory signals. Defective muscle regeneration can severely affect re-innervation by motor axons, and neuromuscular junctions (NMJs) development, ultimately leading to skeletal muscle atrophy. Defects in NMJ formation and maintenance occur physiologically during aging and are responsible for the pathogenesis of several neuromuscular disorders. However, it is still largely unknown how neuromuscular connections are restored on regenerating fibers. It has been suggested that attractive and repelling signals used for axon guidance could be implicated in this process; in particular, guidance molecules called semaphorins play a key role. Semaphorins are a wide family of extracellular regulatory signals with a multifaceted role in cell-cell communication. Originally discovered as axon guidance factors, they have been implicated in cancer progression, embryonal organogenesis, skeletal muscle innervation, and other physiological and developmental functions in different tissues. In particular, in skeletal muscle, specific semaphorin molecules are involved in the restoration and remodeling of the nerve-muscle connections, thus emphasizing their plausible role to ensure the success of muscle regeneration. This review article aims to discuss the impact of aging on skeletal muscle regeneration and NMJs remodeling and will highlight the most recent insights about the role of semaphorins in this context.

DOI: https://doi.org/10.14336/AD.2023.0801
Curr Opin Clin Nutr Metab Care. 2023 Sep 08.

The impact and utility of very low-calorie diets: the role of exercise and protein in preserving skeletal muscle mass.

Tom Anthonius Hubertus Janssen, Derrick W Van Every, Stuart M Phillips.

PURPOSE OF REVIEW: Very low-calorie diets (VLCD) are used as a weight loss intervention, but concerns have been raised about their potential negative impact on lean mass. Here, we review the available evidence regarding the effects of VLCD on lean mass and explore their utility and strategies to mitigate reductions in skeletal muscle.
RECENT FINDINGS: We observed that VLCD, despite their effects on lean mass, may be suitable in certain populations but have a risk in reducing lean mass. The extent of the reduction in lean mass may depend on various factors, such as the duration and degree of energy deficit of the diet, as well as the individual's starting weight and overall health.
SUMMARY: VLCD may be a viable option in certain populations; however, priority needs to be given to resistance exercise training, and secondarily to adequate protein intake should be part of this dietary regime to mitigate losing muscle mass.

DOI: https://doi.org/10.1097/MCO.0000000000000980
Bio Protoc. 2023 Sep 05. 13(17): e4811

Absolute Quantification of mRNA Isoforms in Adult Stem Cells Using Microfluidic Digital PCR.

Shubhangi Das Barman, Zofija Frimand, Antoine De Morree.

  Adult stem cells play key roles in homeostasis and tissue repair. These cells are regulated by a tight control of transcriptional programs. For example, muscle stem cells (MuSCs), located beneath the basal lamina, exist in the quiescent state but can transition to an activated, proliferative state upon injury. The control of MuSC state depends on the expression levels of myogenic transcription factors. Recent studies revealed the presence of different mRNA isoforms, with distinct biological regulation. Quantifying the exact expression levels of the mRNA isoforms encoding these myogenic transcription factors is therefore key to understanding how MuSCs switch between cell states. Previously, quantitative real-time polymerase chain reaction (qRT-PCR) has been used to quantify RNA expression levels. However, qRT-PCR depends on large amounts of RNA input and only measures relative abundance. Here, we present a protocol for the absolute quantification of mRNA isoforms using microfluidic digital PCR (mdPCR). Primary MuSCs isolated from individual skeletal muscles (gastrocnemius and masseter) are lysed, and their RNA is reverse-transcribed into cDNA and copied into double-stranded DNA. Following exonuclease I digestion to remove remaining single-stranded DNA, the samples are loaded onto a mdPCR chip with TaqMan probes targeting the mRNA isoforms of interest, whereupon target molecules are amplified in nanoliter chambers. We demonstrate that mdPCR can give exact molecule counts per cell for mRNA isoforms encoding the myogenic transcription factor Pax3. This protocol enables the absolute quantification of low abundant mRNA isoforms in a fast, precise, and reliable way.

Keywords:  Alternative polyadenylation; Digital PCR; Microfluidics; Muscle stem cells; Pax3; Skeletal muscle; mRNA isoforms

DOI:  https://doi.org/10.21769/BioProtoc.4811
Biomed Pharmacother. 2023 Sep 20. pii: S0753-3322(23)01315-X. [Epub ahead of print]167 115517

Advances in research on cell models for skeletal muscle atrophy.

Liwei Li, Chunman Huang, Jingqun Pang, Yongbin Huang, Xinxin Chen, Guanghua Chen.

  Skeletal muscle, the largest organ in the human body, plays a crucial role in supporting and defending the body and is essential for movement. It also participates in regulating the processes of protein synthesis and degradation. Inhibition of protein synthesis and activation of degradation metabolism can both lead to the development of skeletal muscle atrophy, a pathological condition characterized by a decrease in muscle mass and fiber size. Many physiological and pathological conditions can cause a decline in muscle mass, but the underlying mechanisms of its pathogenesis remain incompletely understood, and the selection of treatment strategies and efficacy evaluations vary. Moreover, the early symptoms of this condition are often not apparent, making it easily overlooked in clinical practice. Therefore, it is necessary to develop and use cell models to understand the etiology and influencing factors of skeletal muscle atrophy. In this review, we summarize the methods used to construct skeletal muscle cell models, including hormone, inflammation, cachexia, genetic engineering, drug, and physicochemical models. We also analyze, compare, and evaluate the various construction and assessment methods.

Keywords:  Cell models; Myotubes; Skeletal muscle atrophy

DOI:  https://doi.org/10.1016/j.biopha.2023.115517
Biofabrication. 2023 Sep 19.

Mimicking sarcolemmal damagein vitro: a contractile 3D model of skeletal muscle for drug testing in Duchenne muscular dystrophy.

Ainoa Tejedera-Villafranca, Marisol Montolio, Javier Ramón-Azcón, Juan M Fernández-Costa.

  Duchenne muscular dystrophy (DMD) is the most prevalent neuromuscular disease diagnosed in childhood. It is a progressive and wasting disease, characterized by a degeneration of skeletal and cardiac muscles caused by the lack of dystrophin protein. The absence of this crucial structural protein leads to sarcolemmal fragility, resulting in muscle fiber damage during contraction. Despite ongoing efforts, there is no cure available for DMD patients. One of the primary challenges is the limited efficacy of current preclinical tools, which fail in modeling the biological complexity of the disease. Human-based 3D cell culture methods appear as a novel approach to accelerate preclinical research by enhancing the reproduction of pathophysiological processes in skeletal muscle. In this work, we developed a patientderived functional 3D skeletal muscle model of DMD that reproduces the sarcolemmal damage found in the native DMD muscle. These bioengineered skeletal muscle tissues exhibit contractile functionality, as they responded to electrical pulse stimulation (EPS). Sustained contractile regimes induced the loss of myotube integrity, mirroring the pathological myotube breakdown inherent in DMD due to sarcolemmal instability. Moreover, damaged DMD tissues showed disease functional phenotypes, such as tetanic fatigue. We also evaluated the therapeutic effect of utrophin upregulator drug candidates on the functionality of the skeletal muscle tissues, thus providing deeper insight into the real impact of these treatments. Overall, our findings underscore the potential of bioengineered 3D skeletal muscle technology to advance DMD research and facilitate the development of novel therapies for DMD and related neuromuscular disorders.

Keywords:  3D cell culture; Duchenne muscular dystrophy; disease modelling; drug testing; sarcolemmal damage; skeletal muscle; tissue engineering

DOI:  https://doi.org/10.1088/1758-5090/acfb3d
Kidney Blood Press Res. 2023 Sep 15.

Irisin ameliorated skeletal muscle atrophy by inhibiting fatty acid oxidation and pyroptosis induced by palmitic acid in chronic kidney disease.

Ting Zhou, Shiyuan Wang, Yajing Pan, Xingtong Dong, Leiyun Wu, Jiali Meng, Jialing Zhang, Qi Pang, Aihua Zhang.

INTRODUCTION: Protein-energy waste (PEW) is a common complication in patients with chronic kidney disease (CKD), among which skeletal muscle atrophy is one of the most important clinical features of PEW. Pyroptosis is a type of proinflammatory programmed cell death associated with skeletal muscle disease. Irisin, as a novel myokine, has attracted extensive attention for its protective role in the complications associated with CKD, but its role in muscle atrophy in CKD is unclear.
METHODS: Palmitic acid (PA) induced muscular atrophy was evaluated by a reduction in C2C12 myotube diameter. Muscle atrophy model was established in male C57BL/6J mice treated with 0.2% adenine for 4 weeks and then fed a 45% high-fat diet.BUN and Cr levels ,body and muscle weight, and muscle histology were assessed. The expression of carnitine palmitoyltransferase 1A (CPT1A) and pyroptosis-related protein was analysed by western blots or immunohistochemistry. The release of IL-1β was detected by ELISA.
RESULTS: In this study, we showed that PA induced muscular atrophy and manifested as a reduction in C2C12 myotube diameter. During this process PA can also induce pyroptosis, as shown by the upregulation of NLRP3, cleaved Caspase1 and GSDMD-N expression and the increased IL-1β release and PI-positive cell rate. Inhibition of Caspase1 or NLRP3 attenuated PA-induced pyroptosis and myotube atrophy in C2C12 cells. Importantly, Irisin treatment significantly ameliorated PA-induced skeletal muscle pyroptosis and atrophy. In terms of mechanism, PA upregulated CPT1A, a key enzyme of fatty acid oxidation(FAO), and Irisin attenuated this effect, which was consistent with Etomoxir (CPT1A inhibitor) treatment. Moreover, Irisin improved skeletal muscle atrophy and pyroptosis in adenine-induced mice by regulating FAO.
CONCLUSION: our study firstly verifies that pyroptosis is a novel mechanism of skeletal muscle atrophy in CKD. Irisin ameliorated skeletal muscle atrophy by inhibiting FAO and pyroptosis in CKD, and Irisin may be developed as a potential therapeutic agent for the treatment of muscle wasting in CKD patients.

DOI: https://doi.org/10.1159/000533926
Cell Metab. 2023 Sep 15. pii: S1550-4131(23)00331-5. [Epub ahead of print]

Regulatory T cells require IL6 receptor alpha signaling to control skeletal muscle function and regeneration.

Maike Becker, Sini S Joseph, Francisco Garcia-Carrizo, Robby Z Tom, Daria Opaleva, Isabelle Serr, Matthias H Tschöp, Tim J Schulz, Susanna M Hofmann, Carolin Daniel.

  Muscle-residing regulatory T cells (Tregs) control local tissue integrity and function. However, the molecular interface connecting Treg-based regulation with muscle function and regeneration remains largely unexplored. Here, we show that exercise fosters a stable induction of highly functional muscle-residing Tregs with increased expression of amphiregulin (Areg), EGFR, and ST2. Mechanistically, we find that mice lacking IL6Rα on T cells (TKO) harbor significant reductions in muscle Treg functionality and satellite and fibro-adipogenic progenitor cells, which are required for muscle regeneration. Using exercise and sarcopenia models, IL6Rα TKO mice demonstrate deficits in Tregs, their functional maturation, and a more pronounced decline in muscle mass. Muscle injury models indicate that IL6Rα TKO mice have significant disabilities in muscle regeneration. Treg gain of function restores impaired muscle repair in IL6Rα TKO mice. Of note, pharmacological IL6R blockade in WT mice phenocopies deficits in muscle function identified in IL6Rα TKO mice, thereby highlighting the clinical implications of the findings.

Keywords:  IL6Ra signaling; exercise; immune tissue crosstalk; immune-metabolic crosstalk; immunometabolism; injury; muscle function; niche-specific Tregs; tissue Tregs; voluntary wheel running

DOI:  https://doi.org/10.1016/j.cmet.2023.08.010
J Cachexia Sarcopenia Muscle. 2023 Sep 21.

Stimulator of interferon genes promotes diabetic sarcopenia by targeting peroxisome proliferator activated receptors γ degradation and inhibiting fatty acid oxidation.

Sen-Bo Yan, Huan Liang, Peng Zhan, Hui Zheng, Qin-Xiao Zhao, Zi-Jie Zheng, Hui-Xia Lu, Guo-Kai Shang, Xiao-Ping Ji.

   BACKGROUND: Declined skeletal muscle mass and function are inevitable consequences of long-term diabetes and bring about many adverse events. Muscle fibre atrophy and interstitial fibrosis are major pathological manifestations of diabetic sarcopenia. Stimulator of interferon genes (STING) participates in various metabolic diseases. We aimed to explore whether and how STING regulates the above pathological manifestations of diabetic sarcopenia.
METHODS: Wild-type and STINGgt/gt C57BL/6J mice and C2C12 myotubes were used to study the role of STING in the regulation of diabetic sarcopenia and the underlying mechanisms.
RESULTS: STING was abnormally activated in diabetic muscles and in PA-treated myotubes (P < 0.01 for all parameters). The diabetic mice demonstrated decreased forelimb grip strength, lean mass, muscle weight and hanging impulse, which were improved by STING deficiency due to alleviated muscle fibre atrophy and interstitial fibrosis (P < 0.05 for all parameters). STING deficiency alleviated muscle fibre atrophy through the following mechanisms. Firstly, STING deficiency or inhibition increased the contents of pDRP1Ser616 , PINK1, Parkin and LC3-II, decreased p62 content, and increased the amount of mito-Keima fluorescent dots at 578 nm in diabetic state (P < 0.05 for all parameters), suggesting improved mitofission and mitophagy. Secondly, STING deficiency or inhibition increased the expression of pAKTSer473 and GLUT4 post-insulin change in diabetic state (P < 0.05 for all), indicating alleviated insulin resistance (IR). Mechanically, STING deficiency or inhibition increased peroxisome proliferator activated receptors γ (PPARγ) protein content by reducing the degradation of ubiquitinated PPARγ through the proteasome pathway and thus increased the expression of fatty acid oxidation (FAO)-related proteins in diabetic state (P < 0.05 for all parameters). Decreased expression of FAO-related proteins caused by PPARγ inhibition abolished the improvements in mitofission, mitophagy and IR achieved by STING inhibition in PA-treated myotubes and thus promoted muscle fibre atrophy (P < 0.05 for all parameters). STING deficiency alleviated interstitial fibrosis by decreasing TGFβ1 expression in diabetic state and TGFβ1 promoted the fibrogenic differentiation of fibro-adipogenic progenitors (P < 0.05 for all parameters). PPARγ inhibition abolished the effect of STING inhibition on reducing TGFβ1 content in PA-treated myotubes (P < 0.01).
CONCLUSIONS: STING deficiency exerted protective effects in diabetic sarcopenia by inhibiting the degradation of ubiquitinated PPARγ through the proteasome pathway and enhancing PPARγ-mediated FAO, which alleviated muscle fibre atrophy by promoting mitophagy and ameliorating IR, and alleviated interstitial fibrosis by reducing TGFβ1 production and suppressing the fibrogenic differentiation of fibro-adipogenic progenitors.

Keywords:  Atrophy; Diabetic sarcopenia; Fibrosis; PPARγ; STING

DOI:  https://doi.org/10.1002/jcsm.13336
Open Biol. 2023 Sep;13(9): 230037

Migration and differentiation of muscle stem cells are coupled by RhoA signalling during regeneration.

Mirco Brondolin, Dylan Herzog, Sami Sultan, Fiona Warburton, Alessandra Vigilante, Robert D Knight.

  Skeletal muscle is highly regenerative and is mediated by a population of migratory adult muscle stem cells (muSCs). Effective muscle regeneration requires a spatio-temporally regulated response of the muSC population to generate sufficient muscle progenitor cells that then differentiate at the appropriate time. The relationship between muSC migration and cell fate is poorly understood and it is not clear how forces experienced by migrating cells affect cell behaviour. We have used zebrafish to understand the relationship between muSC cell adhesion, behaviour and fate in vivo. Imaging of pax7-expressing muSCs as they respond to focal injuries in trunk muscle reveals that they migrate by protrusive-based means. By carefully characterizing their behaviour in response to injury we find that they employ an adhesion-dependent mode of migration that is regulated by the RhoA kinase ROCK. Impaired ROCK activity results in reduced expression of cell cycle genes and increased differentiation in regenerating muscle. This correlates with changes to focal adhesion dynamics and migration, revealing that ROCK inhibition alters the interaction of muSCs to their local environment. We propose that muSC migration and differentiation are coupled processes that respond to changes in force from the environment mediated by RhoA signalling.

Keywords:  cell motility; mechanotransduction; myogenesis; satellite cell; zebrafish

DOI:  https://doi.org/10.1098/rsob.230037
Am J Physiol Endocrinol Metab. 2023 Sep 20.

Both moderate- and high-intensity exercise training increase intramyocellular lipid droplet abundance and modify myocellular distribution in adults with obesity.

Michael W Schleh, Cheehoon Ahn, Benjamin J Ryan, Olivia K Chugh, Austin T Luker, Kathryn E Luker, Jenna B Gillen, Alison C Ludzki, Douglas W Van Pelt, Lisa M Pitchford, Tao Zhang, Thomas Rode, Suzette M Howton, Charles F Burant, Jeffrey F Horowitz.

  Exercise training modifies lipid metabolism in skeletal muscle, but the effect of exercise training on intramyocellular lipid droplet (LD) abundance, size, and intracellular distribution in adults with obesity remains elusive. This study compared high-intensity interval training (HIIT) with more conventional moderate-intensity continuous training (MICT) on intramyocellular lipid content, as well as LD characteristics (size and number) and abundance within the intramyofibrillar (IMF) and subsarcolemmal (SS) regions of type I and type II skeletal muscle fibers in adults with obesity. Thirty-six adults with obesity (BMI=33±3 kg/m2) completed 12 weeks (4d/week) of either HIIT (10x1 min, 90% HRmax + 1 min active recovery; n=19) or MICT (45 min steady-state exercise, 70% HRmax; n=17), while on a weight-maintaining diet throughout training. Skeletal muscle biopsies were collected from the vastus lateralis before and after training, and intramyocellular lipid content and intracellular LD distribution were measured by immunofluorescence microscopy. Both MICT and HIIT increased total intramyocellular lipid content by more than 50% (p<0.01), which was attributed to a greater LD number per µm2 in the IMF region of both type I and type II muscle fibers (p<0.01). Our findings also suggest that LD lipophagy (autophagy-mediated LD degradation) may be transiently upregulated the day after the last exercise training session (p<0.02 for both MICT and HIIT). In summary, exercise programs for adults with obesity involving either MICT or HIIT increased skeletal muscle LD abundance via a greater number of LDs in the IMF region of the myocyte, thereby providing more lipid in close proximity to the site of energy production during exercise.

Keywords:  exercise; high-intensity interval training; lipid droplet; lipophagy; skeletal muscle

DOI:  https://doi.org/10.1152/ajpendo.00093.2023
J Endocrinol. 2023 Sep 01. pii: JOE-23-0210. [Epub ahead of print]

Loss of Trim28 in muscle alters mitochondrial signalling but not systemic metabolism.

Emily J King, Simon T Bond, Christine Yang, Yingying Liu, Anna C Calkin, Darren C Henstridge, Brian G Drew.

Type 2 diabetes mellitus (T2DM), a condition characterised by insulin resistance (IR) and skeletal muscle mitochondrial abnormalities, is a leading cause of death in developed societies. Much work has postulated that improving pathways linked to mitochondrial health, including autophagy, may be a potential avenue to prevent or treat T2DM. Given recent data indicating a role for Tripartite motif-containing 28 (TRIM28) in autophagy and mitochondrial pathways, we investigated whether muscle specific deletion of TRIM28 might impact on obesity, glucose tolerance and IR in mice. We studied two different muscle-specific (MCK-cre and ACTA1-cre-ERT2) TRIM28 knock out (KO) models, which were phenotyped during and after being fed a chow or high fat diet (HFD). Whilst muscle specific deletion of TRIM28 in both models demonstrated alterations in markers of mitochondrial activity and autophagy in skeletal muscle, we did not observe major impacts on the majority of metabolic measures in these mice. Specifically, we demonstrate that deletion of TRIM28 in skeletal muscle of mice during (MCK-cre) or post-development (ACTA1-cre-ERT2) does not prevent HFD-induced obesity or glucose intolerance. These findings are consistent with those reported previously in relation to autophagy and mitochondria in other cell types, and thus warrant further study into the biological role Trim28 has in relation to mitochondrial function.

DOI: https://doi.org/10.1530/JOE-23-0210
Arch Biochem Biophys. 2023 Sep 16. pii: S0003-9861(23)00258-8. [Epub ahead of print] 109759

Diosgenin prevents dexamethasone-induced myotube atrophy in C2C12 cells.

Yasukiyo Yoshioka, Shiori Oishi, Keita Onoda, Katsumi Shibata, Noriyuki Miyoshi.

  Several pathophysiological abnormalities, including a sedentary lifestyle, chronic diseases, and oxidative stress, can contribute to muscle atrophy triggered by an imbalance in muscle protein synthesis and degradation. Resolving muscle atrophy is a critical issue as it can reduce the quality of life. Here, one of the promising functional food factors, diosgenin (a steroidal sapogenin) showed strong preventive activities against dexamethasone (Dex)-induced muscle atrophy, as determined by the expression levels and morphology of the myosin heavy chain in C2C12 myotubes. Diosgenin inhibited protein expressions of Dex-induced skeletal muscle-specific ubiquitin ligase, including muscle RING finger 1 (MuRF1) and casitas B-lineage lymphoma protooncogene b (Cbl-b) but not atrogin-1. Diosgenin ameliorated Dex-induced declines of Akt phosphorylation at Ser473 and FoxO3a phosphorylation at Ser253, which probably at least partially contributed to the suppression of MuRF1, Cbl-b, and atrogin-1 gene expression. Additionally, diosgenin inhibited Dex-induced nuclear translocation of the glucocorticoid receptor (GR), diosgenin therefore may competitively inhibit the interaction between Dex and GR. These findings suggest that diosgenin is an effective functional food for preventing glucocorticoid-induced skeletal muscle atrophy.

Keywords:  C2C12 myotube cells; Dexamethasone; Diosgenin; Glucocorticoid receptor; Muscle atrophy

DOI:  https://doi.org/10.1016/j.abb.2023.109759
Free Radic Biol Med. 2023 Sep 18. pii: S0891-5849(23)00642-1. [Epub ahead of print]

Exhaustive exercise alters native and site-specific H2O2 emission in red and white skeletal muscle mitochondria.

Collins Kamunde, Yashodya Wijayakulathilake, Chidozie Okoye, Nirmala Chinnappareddy, Zahra Kalvani, Michael van den Heuvel, Ravinder Sappal, Don Stevens.

  Mitochondrial reactive oxygen species (ROS) homeostasis is intricately linked to energy conversion reactions and entails regulation of the mechanisms of ROS production and removal. However, there is limited understanding of how energy demand modulates ROS balance. Skeletal muscle experiences a wide range of energy demand depending on the intensity and duration of exercise and therefore is an excellent model to probe the effect of altered energy demand on mitochondrial ROS production. Because in most fish skeletal muscle exists essentially as pure spatially distinct slow-twitch red oxidative and fast-twitch white glycolytic fibers, it provides a natural system for investigating how functional specialization affects ROS homeostasis. We tested the hypothesis that acute increase in energy demand imposed by exhaustive exercise will increase mitochondrial H2O2 emission to a greater extent in red muscle mitochondria (RMM) compared with white muscle mitochondria (WMM). We found that native H2O2 emission rates varied by up to 6-fold depending on the substrate being oxidized and muscle fiber type, with RMM emitting at higher rates with glutamate-malate and palmitoylcarnitine while WMM emitted at higher rates with succinate and glyceral-3-phosphate. Exhaustive exercise increased the native and site-specific H2O2 emission rates; however, the maximal emission rates depended on the substrate, fiber type and redox site. The H2O2 consumption capacity and activities of individual antioxidant enzymes including the glutathione- and thioredoxin-dependent peroxidases as well as catalase were higher in RMM compared with WMM indicating that the activity of antioxidant defense system does not explain the differences in H2O2 emission rates in RMM and WMM. Overall, our study suggests that substrate selection and oxidation may be the key factors determining the rates of ROS production in RMM and WMM following exhaustive exercise.

Keywords:  Antioxidant capacity; Exhaustive exercise; H(2)O(2) emission and consumption; Mitochondria; Red and white skeletal muscle

DOI:  https://doi.org/10.1016/j.freeradbiomed.2023.09.018
Cell Death Dis. 2023 09 18. 14(9): 612

KLF4 regulates skeletal muscle development and regeneration by directly targeting P57 and Myomixer.

Shufang Cai, Xiaoyu Wang, Rong Xu, Ziyun Liang, Qi Zhu, Meilin Chen, Zhuhu Lin, Chenggan Li, Tianqi Duo, Xian Tong, Enru Li, Zuyong He, Xiaohong Liu, Yaosheng Chen, Delin Mo.

Krüppel-like factor 4 (KLF4) is an evolutionarily conserved zinc finger-containing transcription factor that regulates diverse cellular processes such as cell proliferation, apoptosis, and differentiation. Our previous study showed that KLF4 expression is upregulated in skeletal muscle ontogeny during embryonic development in pigs, suggesting its importance for skeletal muscle development and muscle function. We revealed here that KLF4 plays a critical role in skeletal muscle development and regeneration. Specific knockout of KLF4 in skeletal muscle impaired muscle formation further affecting physical activity and also defected skeletal muscle regeneration. In vitro, KLF4 was highly expressed in proliferating myoblasts and early differentiated cells. KLF4 knockdown promoted myoblast proliferation and inhibited myoblast fusion, while its overexpression showed opposite results. Mechanically, in proliferating myoblasts, KLF4 inhibits myoblast proliferation through regulating cell cycle arrest protein P57 by directly targeting its promoter; while in differentiated myoblasts, KLF4 promotes myoblast fusion by transcriptionally activating Myomixer. Our study provides mechanistic information for skeletal muscle development, reduced muscle strength and impaired regeneration after injury and unveiling the mechanism of KLF4 in myogenic regulation.

DOI: https://doi.org/10.1038/s41419-023-06136-w
JCI Insight. 2023 09 22. pii: e171878. [Epub ahead of print]8(18):

Muscle: an independent contributor to the neuromuscular spinal muscular atrophy disease phenotype.

Narendra N Jha, Jeong-Ki Kim, Yoon-Ra Her, Umrao R Monani.

Spinal muscular atrophy (SMA) is a pediatric-onset neuromuscular disorder caused by insufficient survival motor neuron (SMN) protein. SMN restorative therapies are now approved for the treatment of SMA; however, they are not curative, likely due to a combination of imperfect treatment timing, inadequate SMN augmentation, and failure to optimally target relevant organs. Here, we consider the implications of imperfect treatment administration, focusing specifically on outcomes for skeletal muscle. We examine the evidence that muscle plays a contributing role in driving neuromuscular dysfunction in SMA. Next, we discuss how SMN might regulate the health of myofibers and their progenitors. Finally, we speculate on therapeutic outcomes of failing to raise muscle SMN to healthful levels and present strategies to restore function to this tissue to ensure better treatment results.

DOI: https://doi.org/10.1172/jci.insight.171878
Physiol Rep. 2023 Sep;11(18): e15817

Acyl-CoA synthetase expression in human skeletal muscle is reduced in obesity and insulin resistance.

Margarete Poppelreuther, Anne-Marie Lundsgaard, Pernille Mensberg, Kim Sjøberg, Tina Vilsbøll, Bente Kiens, Joachim Füllekrug.

  Upon intramuscular entry, fatty acids are converted to amphiphatic fatty acyl-CoAs by action of the acyl-CoA synthetase (ACS) enzymes. While it has been reported that insulin resistant skeletal muscle shows an accumulation of fatty acyl-CoAs, the role of the enzymes which catalyze their synthesis is still sparsely studied in human muscle, in particular the influence of obesity, and insulin resistance. We analyzed muscle biopsies obtained from normal weight controls (n = 7, average BMI 24), males/females with obesity (n = 7, average BMI 31), and males/females with obesity and type 2 diabetes (T2D) (n = 7, average BMI 34), for relevant ACS (long-chain acyl-CoA synthetase 1 (ACSL1), -3 (ACSL3) and - 4 (ACSL4), fatty acid transport protein 1 (FATP1) and - 4 (FATP4)). The mRNA expression was determined by real-time PCR, and total oleoyl-CoA synthetase activity was measured. In the males/females with obesity and T2D, the response to 16 weeks of exercise training with minor weight loss was evaluated. ACSL1 is the dominantly expressed ACS isoform in human skeletal muscle. The content of total ACS mRNA, as well as ACSL1 mRNA, were lower in muscle of males/females with obesity and T2D. Exercise training in the males/females with obesity and T2D increased the total ACS enzyme activity, along with a lowering of the HOMA-IR index. The capacity for synthesis of fatty acyl-CoAs is lower in skeletal muscle of obese males/females with T2D. This suggests a decreased ability to convert fatty acids to fatty acyl-CoAs, which in turn may affect their entry into storage or metabolic pathways in muscle. Thus, the accumulation of fatty acyl-CoAs in the obese or insulin resistant state that has been shown in previous reports is not likely to result from increased fatty acid acylation. The upregulation of ACS activity by exercise training appears beneficial and occurred concomitantly with increased insulin sensitivity.

Keywords:  ACSL1; fatty acyl-CoA synthetase activity; human skeletal muscle; insulin resistance; type 2 diabetes

DOI:  https://doi.org/10.14814/phy2.15817
Maturitas. 2023 Sep 07. pii: S0378-5122(23)00450-4. [Epub ahead of print]178 107844

The role of estrogen in female skeletal muscle aging: A systematic review.

Annabel J Critchlow, Danielle Hiam, Ross Williams, David Scott, Séverine Lamon.

  Aging is associated with a loss of skeletal muscle mass and function that negatively impacts the independence and quality of life of older individuals. Females demonstrate a distinct pattern of muscle aging compared to males, potentially due to menopause, when the production of endogenous sex hormones declines. This systematic review aims to investigate the current knowledge about the role of estrogen in female skeletal muscle aging. A systematic search of MEDLINE Complete, Global Health, Embase, PubMed, SPORTDiscus, and CINHAL was conducted. Studies were considered eligible if they compared a state of estrogen deficiency (e.g. postmenopausal females) or supplementation (e.g. estrogen therapy) to normal estrogen conditions (e.g. premenopausal females or no supplementation). Outcome variables of interest included measures of skeletal muscle mass, function, damage/repair, and energy metabolism. Quality assessment was completed with the relevant Johanna Briggs critical appraisal tool, and data were synthesized in a narrative manner. Thirty-two studies were included in the review. Compared to premenopausal women, postmenopausal women had reduced muscle mass and strength, but the effect of menopause on markers of muscle damage and expression of the genes involved in metabolic signaling pathways remains unclear. Some studies suggest a beneficial effect of estrogen therapy on muscle size and strength, but evidence is largely conflicting and inconclusive, potentially due to large variations in the reporting and status of exposure and outcomes. The findings from this review point toward a potential negative effect of estrogen deficiency on aging skeletal muscle, but further mechanistic evidence is needed to clarify its role.

Keywords:  Aging; Menopause; Ovarian hormones; Skeletal muscle

DOI:  https://doi.org/10.1016/j.maturitas.2023.107844
Physiol Rep. 2023 Sep;11(18): e15815

Stimulation of fat oxidation in rat muscle by unacylated ghrelin persists for 2-3 hours, but is independent of fatty acid transporter translocation.

Evan M Hoecht, Joshua M Budd, Nicole M Notaro, Graham P Holloway, David J Dyck.

  While a definitive mechanism-of-action remains to be identified, recent findings indicate that ghrelin, particularly the unacylated form (UnAG), stimulates skeletal muscle fatty acid oxidation. The biological importance of UnAG-mediated increases in fat oxidation remains unclear, as UnAG peaks in the circulation before mealtimes, and decreases rapidly during the postprandial situation before increases in postabsorptive circulating lipids. Therefore, we aimed to determine if the UnAG-mediated stimulation of fat oxidation would persist long enough to affect the oxidation of meal-derived fatty acids, and if UnAG stimulated the translocation of fatty acid transporters to the sarcolemma as a mechanism-of-action. In isolated soleus muscle strips from male rats, short-term pre-treatment with UnAG elicited a persisting stimulus on fatty acid oxidation 2 h after the removal of UnAG. UnAG also caused an immediate phosphorylation of AMPK, but not an increase in plasma membrane FAT/CD36 or FABPpm. There was also no increase in AMPK signaling or increased FAT/CD36 or FABPpm content at the plasma membrane at 2 h which might explain the sustained increase in fatty acid oxidation. These findings confirm UnAG as a stimulator of fatty acid oxidation and provide evidence that UnAG may influence the handling of postprandial lipids. The underlying mechanisms are not known.

Keywords:  FABPpm; FAT/CD36; fatty acid oxidation; skeletal muscle; unacylated ghrelin

DOI:  https://doi.org/10.14814/phy2.15815
BMC Cancer. 2023 Sep 20. 23(1): 889

Highlighting the idea of exerkines in the management of cancer patients with cachexia: novel insights and a critical review.

Amirhossein Ahmadi Hekmatikar, André Nelson, Aaron Petersen.

   BACKGROUND: Exerkines are all peptides, metabolites, and nucleic acids released into the bloodstream during and after physical exercise. Exerkines liberated from skeletal muscle (myokines), the heart (cardiokines), liver (hepatokines), white adipose tissue (adipokines), brown adipose tissue (batokines), and neurons (neurokines) may benefit health and wellbeing. Cancer-related cachexia is a highly prevalent disorder characterized by weight loss with specific skeletal muscle and adipose tissue loss. Many studies have sought to provide exercise strategies for managing cachexia, focusing on musculoskeletal tissue changes. Therefore, understanding the responses of musculoskeletal and other tissue exerkines to acute and chronic exercise may provide novel insight and recommendations for physical training to counteract cancer-related cachexia.
METHODS: For the purpose of conducting this study review, we made efforts to gather relevant studies and thoroughly discuss them to create a comprehensive overview. To achieve this, we conducted searches using appropriate keywords in various databases. Studies that were deemed irrelevant to the current research, not available in English, or lacking full-text access were excluded. Nevertheless, it is important to acknowledge the limited amount of research conducted in this specific field.
RESULTS: In order to obtain a comprehensive understanding of the findings, we prioritized human studies in order to obtain results that closely align with the scope of the present study. However, in instances where human studies were limited or additional analysis was required to draw more robust conclusions, we also incorporated animal studies. Finally, 295 studies, discussed in this review.
CONCLUSION: Our understanding of the underlying physiological mechanisms related to the significance of investigating exerkines in cancer cachexia is currently quite basic. Nonetheless, this demonstrated that resistance and aerobic exercise can contribute to the reduction and control of the disease in individuals with cancer cachexia, as well as in survivors, by inducing changes in exerkines.

Keywords:  Cancer-related cachexia; Exercise; Exerkines; Physical exercise

DOI:  https://doi.org/10.1186/s12885-023-11391-3
J Cachexia Sarcopenia Muscle. 2023 Sep 20.

Modelling three-dimensional cancer-associated cachexia and therapy: The molecular basis and therapeutic potential of interleukin-6 transignalling blockade.

Marianna Cosentino, Laura Forcina, Mariam Zouhair, Ludovica Apa, Desirèe Genovese, Caterina Boccia, Emanuele Rizzuto, Antonio Musarò.

   BACKGROUND: Causes and mechanisms underlying cancer cachexia are not fully understood, and currently, no therapeutic approaches are available to completely reverse the cachectic phenotype. Interleukin-6 (IL-6) has been extensively described as a key factor in skeletal muscle physiopathology, exerting opposite roles through different signalling pathways.
METHODS: We employed a three-dimensional ex vivo muscle engineered tissue (X-MET) to model cancer-associated cachexia and to study the effectiveness of selective inhibition of IL-6 transignalling in counteracting the cachectic phenotype. Conditioned medium (CM) derived from C26 adenocarcinoma cells was used as a source of soluble factors contributing to the establishment of cancer cachexia in the X-MET model. A dose of 1.2 ng/mL of glycoprotein-130 fused chimaera (gp130Fc) was added to cachectic culture medium to neutralize IL-6 transignalling.
RESULTS: C26-conditioned medium induced a cachectic-like phenotype in the X-MET, leading to a decline of muscle mass (-60%; P < 0.001), a reduction in myosin expression (-92.4%; P < 0.005) and a reduction of the contraction frequency spectrum (-94%). C26-conditioned medium contains elevated amounts of IL-6 (8.61 ± 4.09 pg/mL) and IL6R (56.85 ± 10.96 pg/mL). These released factors activated the signal transducer and activator of transcription 3 (STAT3) signalling in the C26_CM X-MET system (phosphorylated STAT3/TOTAL +54.6%; P < 0.005), which in turn promote an enhancement of Il-6 (+69.2%; P < 0.05) and Il6r (+43%; P < 0.05) gene expression, suggesting the induction of a feed-forward loop. The selective neutralization of IL-6 transignalling, by gp130Fc, in C26_CM X-MET prevented the hyperactivation of STAT3 (-55.8%; P < 0.005), countered the reduction of cross-sectional area (+28.2%; P < 0.05) and reduced the expression of proteolytic factors including muscle ring finger-1 (-88%; P < 0.005) and ATROGIN1 (-92%; P < 0.05), thus preserving the robustness and increasing the contractile force (+20%) of the three-dimensional muscle system. Interestingly, the selective inhibition of IL-6 transignalling modulated gene regulatory networks involved in myogenesis and apoptosis, normalizing the expression of pro-apoptotic miRNAs, including miR-31 (-53.2%; P < 0.05) and miR-34c (-65%; P < 0.005), and resulting in the reduction of apoptotic pathways highlighted by the sensible reduction of cleaved caspase 3 (-92.5%; P < 0.005) in gp130Fc-treated C26_CM X-MET.
CONCLUSIONS: IL-6 transignalling appeared as a promising target to counter cancer cachexia-related alterations. The X-MET model has proven to be a reliable drug-screening tool to identify novel therapeutic approaches and to test them in preclinical studies, significantly reducing the use of animal models.

Keywords:  IL-6 transignalling blockade; cancer cachexia; drug screening; muscle wasting; three-dimensional skeletal muscle model

DOI:  https://doi.org/10.1002/jcsm.13329
Proc Natl Acad Sci U S A. 2023 Sep 26. 120(39): e2220556120

Iditarod, a Drosophila homolog of the Irisin precursor FNDC5, is critical for exercise performance and cardiac autophagy.

Tyler Cobb, Irene Hwang, Michael Soukar, Sim Namkoong, Uhn-Soo Cho, Maryam Safdar, Myungjin Kim, Robert J Wessells, Jun Hee Lee.

  Mammalian FNDC5 encodes a protein precursor of Irisin, which is important for exercise-dependent regulation of whole-body metabolism. In a genetic screen in Drosophila, we identified Iditarod (Idit), which shows substantial protein homology to mouse and human FNDC5, as a regulator of autophagy acting downstream of Atg1/Atg13. Physiologically, Idit-deficient flies showed reduced exercise performance and defective cold resistance, which were rescued by exogenous expression of Idit. Exercise training increased endurance in wild-type flies, but not in Idit-deficient flies. Conversely, Idit is induced upon exercise training, and transgenic expression of Idit in wild-type flies increased endurance to the level of exercise trained flies. Finally, Idit deficiency prevented both exercise-induced increase in cardiac Atg8 and exercise-induced cardiac stress resistance, suggesting that cardiac autophagy may be an additional mechanism by which Idit is involved in the adaptive response to exercise. Our work suggests an ancient role of an Iditarod/Irisin/FNDC5 family of proteins in autophagy, exercise physiology, and cold adaptation, conserved throughout metazoan species.

Keywords:  FNDC5; Irisin; autophagy; cardiac; exercise

DOI:  https://doi.org/10.1073/pnas.2220556120
J Physiol. 2023 Sep 19.

The separation and identification of circulating small extracellular vesicles from endurance-trained, strength-trained and recreationally active men.

Ian A J Darragh, Niamh McNamee, Róisín Daly, Sarai Martinez Pacheco, Lorraine O'Driscoll, Brendan Egan.

  Small extracellular vesicles (EV) are membrane-encapsulated particles that carry bioactive cargoes, are released by all cell types and are present in all human biofluids. Changes in EV profiles and abundance occur in response to acute exercise, but this study investigated whether individuals with divergent histories of exercise training (recreationally active controls - CON; endurance-trained - END; strength-trained - STR) presented with varied abundances of small EVs in resting samples and whether the abundance of small EVs differed within each group across two measurement days. Participants (n = 38, all male; CON n = 12, END n = 13, STR n = 13) arrived at the lab on two separate occasions in a rested, overnight fasted state, with standardisation of time of day of sampling, recent dietary intake, time since last meal and time since last exercise training session (∼40 h). Whole blood samples were collected and separated into plasma from which small EVs were separated using size exclusion chromatography and identified in accordance with the Minimal Information For Studies of Extracellular Vesicles (MISEV) guidelines. No differences in the abundance of small EVs were observed within or between groups across multiple methods of small EV identification (nanoparticle tracking analysis, flow cytometry, immunoblot of specific EV markers). Targeted metabolomics of the small EV preparations identified 96 metabolites that were associated with the structure and function of small EVs, with no statistically significant differences in concentrations observed across groups. The results of the current study suggest that the abundance and metabolomic profile of small EVs derived from men with divergent histories of exercise training are similar to those in resting blood samples. KEY POINTS: Extracellular vesicles (EV) are membrane-encapsulated particles that are present in circulation and carry bioactive materials as 'cargo'. The abundance and profile of small EVs are responsive to acute exercise, but little is known about the relationship between small EVs and exercise training. This study examined the abundance, and a targeted metabolomic profile, of small EVs separated from the blood of endurance athletes, strength athletes and recreationally active controls at rest (∼40 h after the most recent exercise session) on two separate but identical lab visits. No differences were observed in the abundance or metabolomic profile of small EV preparations between the groups or between the lab visits within each group. Further research should determine whether the bioactive cargoes (e.g. RNA, protein and additional metabolites) carried within EVs are altered in individuals with divergent histories of exercise training or in response to exercise training interventions.

Keywords:  athletes; exercise training; exosome; metabolomics

DOI:  https://doi.org/10.1113/JP285170
Mediators Inflamm. 2023 ;2023 9018470

Influence of Inflammatory Cytokines IL-1β and IFNγ on Sarcoplasmic Aggregation of p62 and TDP-43 in Myotubes.

Bryony McCord, Richard M Day.

Skeletal muscle of patients with sporadic inclusion body myositis (sIBM) presents with inflammation, including upregulation of inflammatory cytokines such as interferon γ (IFNγ). Non-inflammatory features are also observed, like the sarcoplasmic accumulation of proteins including TDP-43 and p62. This study aimed to investigate the effect of IFNγ and interleukin 1-β (IL-1β) on TDP-43 and p62 aggregation in vitro. Primary human myotubes were treated with IL-1β (20 ng/mL) and IFNγ (750 ng/mL) separately or combined for 48 hr. Sarcoplasmic TDP-43 aggregates and p62 puncta were assessed using image analysis for size, frequency, and colocalization with each other. Total protein expression of TDP-43, p62 and LC3 was assessed using western blotting. The subcellular localization of TDP-43 was also analyzed using image analysis. Combined IL-1β and IFNγ treatment increased puncta size of p62 compared to control (0.49 ± 0.13 µm2 versus 0.28 ± 0.06 µm2), without affecting puncta frequency or p62 expression but with an increased LC3II/LC3I ratio, suggesting autophagic alterations. IL-1β or IFNγ did not alter p62 puncta size or frequency, suggesting a combined insult of multiple inflammatory mediators is necessary to cause p62 alterations. IL-1β increased p62 protein expression in an autophagy-independent manner. None of the cytokine treatments affected TDP-43 protein expression, size, or frequency of TDP-43 aggregates or localization, suggesting IL-1β and IFNγ may influence TDP-43 processing in human skeletal muscle cells. TDP-43 was localized to the sarcoplasm under control conditions, suggesting this may not be a pathological feature. Overall, sIBM-like TDP-43/p62 features were not triggered by IL-1β and/or IFNγ.

DOI: https://doi.org/10.1155/2023/9018470
J Appl Physiol (1985). 2023 Sep 21.

Effects of fatigue on intra-muscle force-stabilizing synergies.

Joseph M Ricotta, Sayan D Deep De, Mauro Nardon, Anna Benamati, Mark L Latash.

  We applied the recently introduced concept of intra-muscle synergies in spaces of motor units (MUs) to quantify indices of such synergies in the tibialis anterior during ankle dorsiflexion force production tasks and their changes with fatigue. We hypothesized that MUs would be organized into robust groups (MU-modes), which would co-vary across trials to stabilize force magnitude, and the indices of such synergies would drop under fatigue. Healthy, young subjects (n = 14; 7 females) produced cyclical, isometric dorsiflexion forces while surface electromyography was used to identify action potentials of individual MUs. Principal component analysis was used to define MU-modes. The framework of the uncontrolled manifold (UCM) was used to analyze inter-cycle variance and compute the synergy index, ∆VZ. The tests were repeated after a non-fatiguing exercise (control) and fatiguing exercise. Across subjects, fatigue led, on average, to a 43% drop in maximal force and fewer identified MUs per subject (29.6±2.1 vs. 32.4±2.1). The first two MU-modes accounted for 81.2±0.08% of variance across conditions. Force-stabilizing synergies were present across all conditions and was diminished after fatiguing exercise (1.49±0.40), but not control exercise (1.76±0.75). Decreased stability after fatigue was caused by an increase in the amount of variance orthogonal to the UCM. These findings contrast with earlier studies of multi-effector synergies demonstrating increased synergy index under fatigue. We interpret the results as reflections of a drop in the gain of spinal reflex loops under fatigue. The findings corroborate an earlier hypothesis on the spinal nature of intra-muscle synergies.

Keywords:  fatigue; force production; motor unit; synergy; tibialis anterior

DOI:  https://doi.org/10.1152/japplphysiol.00419.2023