Obestatin Treatment Counteracts Muscle Wasting by Reactivation of Autophagy in Duchenne Muscular Dystrophy

Abstract

The mechanisms by which muscular dystrophy-related stress is transduced to the autophagic machinery remain poorly characterized. The formulation of strategies should be based on how disruption of these processes results in the deregulation of signaling pathways that contribute to many pathological effects of the disease. In this study, we investigated the molecular mechanism by which the obestatin/GPR39 system, an autocrine signaling with anabolic impact on normal skeletal muscle, restores autophagy in Duchenne muscular dystrophy (DMD). We report that obestatin integrates 5' AMP-activated protein kinase (AMPK) and mammalian target of rapamycin complex 1 (mTORC1) signaling to control ubiquitin proteasome system (UPS), autophagy–lysosome system, and protein synthesis under dystrophic context. The posttranslational modifications of the E3 ligase NEDD4-L emerges as the main switch to activate the autophagy in response to obestatin. This includes NEDD4-L tyrosine phosphorylation and autoubiquitination, which is critical for recruiting the ubiquitin-specific protease 10 to assemble a deubiquitination complex, that orchestrates the unc-51 like autophagy activating kinase 1 (ULK1) and class III PI3K (VPS34) complexes. Reactivation of autophagy through obestatin signaling promotes the recovery of physiological skeletal muscle function. Thus, DMD conditions determine permissiveness to the activation of AMPK that sustain autophagy under anabolic conditions stablished by obestatin signaling through mTORC1.

Keywords

autophagy / Duchenne muscular dystrophy / neural precursor cell expressed developmentally downregulated protein 4 / obestatin / ubiquitin-specific protease 10

References

[1] S. Crisafulli, J. Sultana, A. Fontana, et al., “Global epidemiology of Duchenne Muscular Dystrophy: An Updated Systematic Review and Meta-Analysis,” Orphanet Journal of Rare Diseases 15, no. 1 (2020): 141.

[2] D. Duan, N. Goemans, S. Takeda, et al., “Duchenne Muscular Dystrophy,” Nature Reviews Disease Primers 7, no. 7 (2021): 13.

[3] M. Koenig, A. P. Monaco, and L. M. Kunkel, “The Complete Sequence of Dystrophin Predicts a Rod-Shaped Cytoskeletal Protein,” Cell 53, no. 53 (1988): 219–228.

[4] J. M. Ervasti and K. P. Campbell, “Membrane Organization of the Dystrophin-glycoprotein Complex,” Cell 66, no. 6 (1991): 1121–1131.

[5] D. G. Allen, O. L. Gervasio, E. W. Yeung, E. W. eYeung, and N. P. Whitehead, “Calcium and the Damage Pathways in Muscular Dystrophy,” Canadian Journal of Physiology and Pharmacology 88, no. 88 (2010): 83–91.

[6] S. Cirak, L. Feng, K. Anthony, et al., “Restoration of the Dystrophin-Associated Glycoprotein Complex After Exon Skipping Therapy in Duchenne Muscular Dystrophy,” Molecular Therapy 20, no. 20 (2012): 462–467.

[7] E. Mercuri, C. G. Bönnemann, and F. Muntoni, “Muscular Dystrophies,” Lancet 394, no. 394 (2019): 2025–2038.

[8] C. De Palma, F. Morisi, and S. Cheli, “Autophagy as a New Therapeutic Target in Duchenne Muscular Dystrophy,” Cell Death & Disease 5, no. 8 (2014): e1363.

[9] C. De Palma, C. Perrotta, P. Pellegrino, E. Clementi, and D. Cervia, “Skeletal Muscle Homeostasis in duchenne Muscular Dystrophy: Modulating Autophagy as a Promising Therapeutic Strategy,” Frontiers in Aging Neuroscience 6 (2014): 188.

[10] E. Fiacco, F. Castagnetti, V. Bianconi, et al., “Autophagy Regulates Satellite Cell Ability to Regenerate Normal and Dystrophic Muscles,” Cell Death and Differentiation 23, no. 11 (2016): 1839–1849.

[11] M. Eagle, J. Bourke, and R. Bullock, “Managing Duchenne Muscular Dystrophy-the Additive Effect of Spinal Surgery and Home Nocturnal Ventilation in Improving Survival,” Neuromuscular Disorders 17, no. 6 (2007): 470–475.

[12] R. T. Moxley 3rd, S. Pandya, E. Ciafaloni, D. J. Fox, and K. Campbell, “Change in Natural History of Duchenne Muscular Dystrophy With Long-term Corticosteroid Treatment: Implications for Management,” Journal of Child Neurology 25, no. 9 (2010): 1116–1129.

[13] T. Saito, M. Kawai, E. Kimura, et al., “Study of Duchenne Muscular Dystrophy Long-term Survivors Aged 40 Years and Older Living in Specialized Institutions in Japan,” Neuromuscular Disorders 27, no. 2 (2017): 107–114.

[14] O. Sheikh and T. Yokota, “Advances in Genetic Characterization and Genotype-Phenotype Correlation of Duchenne and Becker Muscular Dystrophy in the Personalized Medicine Era,” Journal of Personalized Medicine 10 (2020): 111.

[15] D. J. Birnkrant, K. Bushby, C. M. Bann, et al., “DMD Care Considerations Working Group. Diagnosis and Management of Duchenne Muscular Dystrophy, Part 1: Diagnosis, and Neuromuscular, Rehabilitation, Endocrine, and Gastrointestinal and Nutritional Management,” Lancet Neurology 17, no. 3 (2018): 251–267.

[16] K. Bushby, R. Finkel, D. J. Birnkrant, et al., “DMD Care Considerations Working Group. Diagnosis and Management of Duchenne Muscular Dystrophy, Part 1: Diagnosis, and Pharmacological and Psychosocial Management,” Lancet Neurology 9, no. 1 (2010): 77–93.

[17] R. C. Griggs, J. P. Miller, C. R. Greenberg, et al., “Efficacy and Safety of deflazacort vs prednisone and placebo for Duchenne Muscular Dystrophy,” Neurology 87, no. 20 (2016): 2123–2131.

[18] E. Matthews, R. Brassington, T. Kuntzer, and A. Y. Jichi F Manzur, “Corticosteroids for the Treatment of Duchenne Muscular Dystrophy,” Cochrane Database of Systematic Reviews (Online) 5 (2016): CD003725.

[19] W. D. Biggar, A. Skalsky, and C. M. McDonald, “Comparing Deflazacort and Prednisone in Duchenne Muscular Dystrophy,” Journal of Neuromuscular Diseases 9, no. 4 (2022): 463–476.

[20] Q. Q. Gao and E. M. McNally, “The Dystrophin Complex: Structure, Function, and Implications for Therapy,” Comprehensive Physiology 5, no. 3 (2015): 1223–1239.

[21] D. G. Allen, N. P. Whitehead, and S. C. Froehner, “Absence of Dystrophin Disrupts Skeletal Muscle Signaling: Roles of Ca2+, Reactive Oxygen Species, and Nitric Oxide in the Development of Muscular Dystrophy,” Physiological Reviews 96, no. 1 (2016): 253–305.

[22] G. S. Pilgram, S. Potikanond, R. A. Baines, L. G. Fradkin, and J. N. Noordermeer, “The Roles of the Dystrophin-Associated Glycoprotein Complex at the Synapse,” Molecular Neurobiology 41, no. 1 (2010): 1–21.

[23] W. Chen, Y. Chen, Y. Liu, and X. Wang, “Autophagy in Muscle Regeneration: Potential Therapies for Myopathies,” Journal of Cachexia, Sarcopenia & Muscle 13, no. 3 (2022): 1673–1685.

[24] R. Pal, M. Palmieri, J. A. Loehr, et al., “Src-dependent Impairment of Autophagy by Oxidative Stress in a Mouse Model of Duchenne Muscular Dystrophy,” Nature Communications 5 (2014): 4425.

[25] Y. Xiong, Y. Zhou, and H. W. Jarret, “Dystrophin Glycoprotein Complex-associated Gbetagamma Subunits Activate Phosphatidylinositol-3-kinase/Akt Signaling in Skeletal Muscle in a Laminin-dependent Manner,” Journal of Cellular Physiology 219, no. 2 (2009): 402–414.

[26] M. Pauly, F. Daussin, Y. Burelle, et al., “AMPK Activation Stimulates Autophagy and Ameliorates Muscular Dystrophy in the mdx Mouse Diaphragm,” American Journal of Pathology 181, no. 2 (2012): 583–592.

[27] F. Langone, S. Cannata, C. Fuoco, et al., “Metformin Protects Skeletal Muscle From Cardiotoxin Induced Degeneration,” PLoS ONE 9, no. 12 (2014): e114018.

[28] F. Burdet, M. Ibberson, P. Aebischer, et al., “The Mitophagy Activator Urolithin A Is Safe and Induces a Molecular Signature of Improved Mitochondrial and Cellular Health in Humans,” Nature Metabolism 1, no. 6 (2019): 595–603.

[29] P. P. Laurila, M. Wohlwend, H. Li, et al., “Urolithin A Improves Muscle Function by Inducing Mitophagy in Muscular Dystrophy,” Science Translational Medicine 13, no. 588 (2021): eabb0319.

[30] U. Gurriaran-Rodriguez, I. Santos-ZasI, and O. Al-Massadi, “The Obestatin/GPR39 System Is Up-regulated by Muscle Injury and Functions as an Autocrine Regenerative System,” Journal of Biological Chemistry 287, no. 45 (2012): 38379–38389.

[31] U. Gurriaran-Rodriguez, I. Santos-Zas, J. Gonzalez-Sanchez, et al., “Action of Obestatin in Skeletal Muscle Repair: Stem Cell Expansion, Muscle Growth, and Microenvironment Remodeling,” Molecular Therapy 23, no. 6 (2015): 1003–1021.

[32] I. Santos-Zas, U. Gurriarán-Rodríguez, T. Cid-Díaz, et al., “β-Arrestin Scaffolds and Signaling Elements Essential for the Obestatin/GPR39 System That Determine the Myogenic Program in human Myoblast Cells,” Cellular and Molecular Life Sciences 73, no. 3 (2016): 617–635.

[33] I. Santos-Zas, T. Cid-Diaz, J. Gonzalez-Sanchez, et al., “Obestatin Controls Skeletal Muscle fiber-type Determination,” Scientific Reports 7, no. 1 (2017): 2137.

[34] T. Cid-Diaz, I. Santos-Zas, J. Gonzalez-Sanchez, et al., “Obestatin Controls the Ubiquitin-proteasome and Autophagy-lysosome Systems in Glucocorticoid-induced Muscle Cell Atrophy,” Journal of Cachexia, Sarcopenia & Muscle 8, no. 6 (2017): 974–990.

[35] T. Cid-Diaz, S. Leal-Lopez, F. Fernandez-Barreiro, et al., “Obestatin Signalling Counteracts Glucocorticoid-induced Skeletal Muscle Atrophy via NEDD4/KLF15 Axis,” Journal of Cachexia, Sarcopenia & Muscle 12, no. 2 (2021): 493–505.

[36] J. Gonzalez-Sanchez, A. Sanchez-Temprano, T. Cid-Diaz, et al., “Improvement of Duchenne Muscular Dystrophy Phenotype Following Obestatin Treatment,” Journal of Cachexia, Sarcopenia & Muscle 9, no. 6 (2018): 1063–1078.

[37] M. Antonioli, M. Di Rienzo, M. Piacentini, and G. M. Fimia, “Emerging Mechanisms in Initiating and Terminating Autophagy,” Trends in Biochemical Sciences 42, no. 1 (2017): 28–41.

[38] P. Sousa-Victor, L. Garcia-Prat, A. L. Serrano, E. Perdiguero, and P. Muñoz-Cánoves, “Muscle Stem Cell Aging: Regulation and Rejuvenation,” Trends in Endocrinology and Metabolism 26, no. 6 (2015): 287–296.

[39] C. Lennicke and H. M. Cochemé, “Redox Metabolism: ROS as Specific Molecular Regulators of Cell Signaling and Function,” Molecular Cell 81, no. 18 (2021): 3691–3707.

[40] H. Xu and D. Ren, “Lysosomal Physiology,” Annual Review of Physiology 77 (2015): 57–80.

[41] R. Pal, M. Palmieri, J. A. Loehr, et al., “Src-dependent Impairment of Autophagy by Oxidative Stress in a Mouse Model of Duchenne Muscular Dystrophy,” Nature Communications 5 (2014): 4425.

[42] L. Yu, X. Zhang, Y. Yang, et al., “Small-molecule Activation of Lysosomal TRP Channels Ameliorates Duchenne Muscular Dystrophy in Mouse Models,” Science Advances 6, no. 6 (2020): eaaz2736.

[43] S. K. Hindupur, A. Gonzalez, and M. N. Hall, “The Opposing Actions of Target of Rapamycin and AMP-activated Protein Kinase in Cell Growth Control,” Cold Spring Harbor Perspectives in Biology 7 (2015): a019141.

[44] M. Shimobayashi and M. N. Hall, “Multiple Amino Acid Sensing Inputs to mTORC1,” Cell Research 26, no. 1 (2016): 7–20.

[45] N. Hosokawa, T. Sasaki, S. Iemura, T. Natsume, T. Hara, and N. Mizushima, “Atg101, a Novel Mammalian Autophagy Protein Interacting With Atg13,” Autophagy 5, no. 7 (2009): 973–979.

[46] J. Kim, M. Kundu, B. Viollet, and K. L. Guan, “AMPK and mTOR Regulate Autophagy Through Direct Phosphorylation of Ulk1,” Nature Cell Biology 13, no. 2 (2011): 132–141.

[47] D. F. Egan, D. B. Shackelford, M. M. Mihaylova, et al., “Phosphorylation of ULK1 (hATG1) by AMP-activated Protein Kinase Connects Energy Sensing to Mitophagy,” Science 331, no. 6016 (2011): 456–461.

[48] D. F. Egan, M. G. Chun, M. Vamos, et al., “Small Molecule Inhibition of the Autophagy Kinase ULK1 and Identification of ULK1 Substrates,” Molecular Cell 59, no. 2 (2015): 285–297.

[49] R. C. Russell, Y. Tian, and H. Yuan, “ULK1 induces Autophagy by Phosphorylating Beclin-1 and Activating VPS34 Lipid Kinase,” Nature Cell Biology 15, no. 7 (2013): 741–750.

[50] L. Vucicevic, M. Misirkic, K. Janjetovic, et al., “Compound C Induces Protective Autophagy in Cancer Cells Through AMPK Inhibition-independent Blockade of Akt/mTOR Pathway,” Autophagy 7, no. 1 (2011): 40–50.

[51] D. Dalle Pezze, S. Ruf, A. G. Sonntag, et al., “A Systems Study Reveals Concurrent Activation of AMPK and mTOR by Amino Acids,” Nature Communications 7 (2016): 13254.

[52] Y. Xie, R. Kang, X. Sun, et al., “Posttranslational Modification of Autophagy-related Proteins in Macroautophagy,” Autophagy 11, no. 1 (2015): 8–45.

[53] L. Wells, L. K. Kreppel, F. I. Comer, B. E. Wadzinski, and G. W. Hart, “O-GlcNAc Transferase Is in a Functional Complex With Protein Phosphatase 1 Catalytic Subunits,” Journal of Biological Chemistry 279, no. 37 (2004): 38466–38470.

[54] K. E. Pyo, C. R. Kim, M. Lee, J. S. Kim, K. I. Kim, and S. H. Baek, “ULK1 O-GlcNAcylation Is Crucial for Activating VPS34 via ATG14L During Autophagy Initiation,” Cell Reports 25, no. 10 (2018): 2878–2890.e4.

[55] M. Di Rienzo, M. Antonioli, C. Fusco, et al., “Autophagy Induction in Atrophic Muscle Cells Requires ULK1 Activation by TRIM32 Through Unanchored K63-linked Polyubiquitin Chains,” Science Advances 5, no. 5 (2019): eaau8857.

[56] W. Xie, S. Jin, and Y. Wu, “Auto-ubiquitination of NEDD4-1 Recruits USP13 to Facilitate Autophagy Through Deubiquitinating VPS34,” Cell Reports 30, no. 8 (2020): 2807–2819.e4.

[57] M. Deng, X. Yang, B. Qin, et al., “Deubiquitination and Activation of AMPK by USP10,” Molecular Cell 61, no. 4 (2016): 614–624.

[58] L. Buetow and D. T. Huang, “Structural Insights Into the Catalysis and Regulation of E3 Ubiquitin Ligases,” Nature Reviews Molecular Cell Biology 17, no. 10 (2016): 626–642.

[59] P. Y. Jean-Charles, J. C. Snyder, and S. K. Shenoy, “Chapter One—Ubiquitination and Deubiquitination of G Protein-Coupled Receptors,” Progress in Molecular Biology and Translational Science 141 (2016): 1–55.

[60] N. J. Grimsey, R. Narala, and C. C. Rada CC, “A Tyrosine Switch on NEDD4-2 E3 Ligase Transmits GPCR Inflammatory Signaling,” Cell Reports 24, no. 12 (2018): 3312–3323.

[61] J. H. Hurley and L. N. Young, “Mechanisms of Autophagy Initiation,” Annual Review of Biochemistry 86 (2017): 225–244.

[62] X. Ma, P. Li, and L. Ge, “Targeting of Biomolecular Condensates to the Autophagy Pathway,” Trends in Cell Biology 33, no. 6 (2023): 505–516.

[63] J. N. S. Vargas, M. Hamasaki, T. Kawabata, R. J. Youle, and T. Yoshimori, “The Mechanisms and Roles of Selective Autophagy in Mammals,” Nature Reviews Molecular Cell Biology 24, no. 3 (2023): 167–185.

[64] D. G. Hardie, B. Schaffer, and A. Brunet, “AMPK: An Energy-Sensing Pathway With Multiple Inputs and Outputs,” Trends in Cell Biology 26, no. 3 (2016): 190–201.

[65] B. K. Kennedy and D. W. Lamming, “The Mechanistic Target of Rapamycin: The Grand ConducTOR of Metabolism and Aging,” Cell Metabolism 23, no. 6 (2016): 990–1003.

[66] R. A. Saxton and D. M. Sabatini, “mTOR Signaling in Growth, Metabolism, and Disease,” Cell 169, no. 2 (2017): 361–371.

[67] A. Gonzalez, M. N. Hall, S. C. Lin, and D. G. Hardie, “AMPK and TOR: The Yin and Yang of Cellular Nutrient Sensing and Growth Control,” Cell Metabolism 31, no. 3 (2020): 472–492.

[68] G. Y. Liu GY and D. M. Sabatini, “mTOR at the Nexus of Nutrition, Growth, Ageing and Disease,” Nature Reviews Molecular Cell Biology 21, no. 4 (2020): 183–203.

[69] G. R. Steinberg and D. G. Hardie, “New Insights Into Activation and Function of the AMPK,” Nature Reviews Molecular Cell Biology 24, no. 4 (2023): 255–272.

[70] I. Yoon, M. Nam, H. K. Kim, et al., “Glucose-dependent Control of Leucine Metabolism by Leucyl-tRNA Synthetase 1,” Science 367, no. 6474 (2020): 205–210.

[71] L. Shang, S. Chen, F. Du, L. Zhao, and X. Wang, “Nutrient Starvation Elicits an Acute Autophagic Response Mediated by Ulk1 Dephosphorylation and Its Subsequent Dissociation From AMPK,” Proceedings of the National Academy of Sciences of the United States of America 108, no. 12 (2011): 4788–4793.

[72] C. Nwadike, L. E. Williamson, L. E. Gallagher, J. L. Guan, and E. Y. W. Chan, “AMPK Inhibits ULK1-Dependent Autophagosome Formation and Lysosomal Acidification via Distinct Mechanisms,” Molecular and Cellular Biology 38 (2018): e00023–18.

[73] K. R. Morrison, W. J. Smiles, N. X. Y. Lin, et al., “An AMPKα2-specific Phospho-switch Controls Lysosomal Targeting for Activation,” Cell Reports 38, no. 7 (2022): 110365.

[74] H. Xi, J. C. Barredo, J. R. Merchan, and T. J. Lampidis, “Endoplasmic Reticulum Stress Induced by 2-deoxyglucose but Not Glucose Starvation Activates AMPK Through CaMKKβ Leading to Autophagy,” Biochemical Pharmacology 85, no. 10 (2013): 1463–1477.

[75] M. Høyer-Hansen, L. Bastholm, P. Szyniarowski, et al., “Control of Macroautophagy by Calcium, Calmodulin-dependent Kinase Kinase-beta, and Bcl-2,” Molecular Cell 25, no. 2 (2007): 193–205.

[76] D. Vara-Ciruelos, M. Dandapani, A. Gray, E. O. Egbani, A. M. Evans, and D. G. Hardie, “Genotoxic Damage Activates the AMPK-α1 Isoform in the Nucleus via Ca2+/CaMKK2 Signaling to Enhance Tumor Cell Survival,” Molecular Cancer Research 16, no. 2 (2018): 345–357.

[77] S. Li, Z. Lavagnino, D. Lemacon, et al., “Ca2+-Stimulated AMPK-Dependent Phosphorylation of Exo1 Protects Stressed Replication Forks From Aberrant Resection,” Molecular Cell 74, no. 6 (2019): 1123–1137.e6.

[78] N. X. Y. Ling, A. Kaczmarek, A. Hoque, et al., “mTORC1 directly Inhibits AMPK to Promote Cell Proliferation Under Nutrient Stress,” Nature Metabolism 2, no. 1 (2020): 41–49.

[79] S. Kolla, M. Ye, K. G. Mark, and M. Rapé, “Assembly and Function of Branched Ubiquitin Chains,” Trends in Biochemical Sciences 47, no. 9 (2022): 759–771.

[80] M. Deng, X. Yang, B. Qin, et al., “Deubiquitination and Activation of AMPK by USP10,” Molecular Cell 61, no. 4 (2016): 614–624.

[81] J. Xiao, T. Zhang, D. Xu, et al., “FBXL20-mediated Vps34 Ubiquitination as a p53 Controlled Checkpoint in Regulating Autophagy and Receptor Degradation,” Genes & Development 29, no. 2 (2015): 184–196.

[82] C. C. Liu, Y. C. Lin, C. YH, et al., “Cul3-KLHL20 Ubiquitin Ligase Governs the Turnover of ULK1 and VPS34 Complexes to Control Autophagy Termination,” Molecular Cell 61 (2016):84–97.

[83] J. Liu, M. Li, L. Li, S. Chen, and X. Wang, “Ubiquitination of the PI3-kinase VPS-34 Promotes VPS-34 Stability and Phagosome Maturation,” Journal of Cell Biology 217, no. 1 (2018): 347–360.

[84] D. Rotin and S. Kumar, “Physiological Functions of the HECT family of Ubiquitin Ligases,” Nature Reviews Molecular Cell Biology 10, no. 6 (2009): 398–409.

[85] A. Persaud, P. Alberts, S. Mari, et al., “Tyrosine Phosphorylation of NEDD4 Activates Its Ubiquitin Ligase Activity,” Science Signaling 7 (2014): ra95.

[86] M. Margeta, “Autophagy Defects in Skeletal Myopathies,” Annual Review of Pathology 15 (2020): 261–285.

[87] M. C. Vila, S. Rayavarapu, M. W. Hogarth, et al., “Mitochondria Mediate Cell Membrane Repair and Contribute to Duchenne Muscular Dystrophy,” Cell Death and Differentiation 24, no. 2 (2017): 330–342.

[88] X. Hong, J. Isern, S. Campanario, et al., “Mitochondrial Dynamics Maintain Muscle Stem Cell Regenerative Competence throughout Adult Life by Regulating Metabolism and Mitophagy,” Cell Stem Cell 29, no. 9 (2022): 1298–1314.e10.

[89] G. Twig, A. J. M. A. Elorza, and H. Mohamed, “Fission and Selective Fusion Govern Mitochondrial Segregation and Elimination by Autophagy,” Embo Journal 27, no. 2 (2008):433–446.

[90] L. Garcia-Prat, M. Martinez-Vicente, E. Perdiguero, et al., “Autophagy Maintains Stemness by Preventing Senescence,” Nature 529, no. 7584 (2016): 37–42.

[91] A. S. Rambold and E. L. Pearce, “Mitochondrial Dynamics at the Interface of Immune Cell Metabolism and Function,” Trends in Immunology 39, no. 1 (2018): 6–18.

[92] M. Giacomello, A. Pyakurel, C. Glytsou, and L. Scorrano, “The Cell Biology of Mitochondrial Membrane Dynamics,” Nature Reviews Molecular Cell Biology 21, no. 4 (2020): 204–224.

[93] R. Kodama, M. Kato, S. Furuta, et al., “ROS-generating Oxidases Nox1 and Nox4 Contribute to Oncogenic Ras-induced Premature Senescence,” Genes to Cells 18, no. 1 (2013): 32–41.

[94] C. Lopez-Otin, M. A. Blasco, L. Partridge, M. Serrano, and G. Kroemer, “The Hallmarks of Aging,” Cell 153, no. 6 (2013): 1194–1217.

[95] Y. Endo, A. Furuta, and I. Nishino, “Danon Disease: A Phenotypic Expression of LAMP-2 Deficiency,” Acta Neuropathologica 129, no. 3 (2015): 391–398.

[96] L. Kohler, R. Puertollano, and N. Raben, “Pompe Disease: From Basic Science to Therapy,” Neurotherapeutics 15, no. 4 (2018): 928–942.

[97] P. Grumati, L. Coletto, P. Sabatelli, et al., “Autophagy Is Defective in Collagen VI Muscular Dystrophies, and Its Reactivation Rescues Myofiber Degeneration,” Nature Medicine 16, no. 11 (2010):1313–1320.

[98] P. Castets, S. Frank, M. Sinnreich, and M. A. Rüegg, “Get the Balance Right': Pathological Significance of Autophagy Perturbation in Neuromuscular Disorders,” Journal of Neuromuscular Diseases 3, no. 2 (2016): 127–155.

[99] R. E. Lawrence and R. Zoncu, “The Lysosome as a Cellular Centre for Signalling, Metabolism and Quality Control,” Nature Cell Biology 21, no. 2 (2019): 133–142.

[100] D. G. S. Wilson, A. Tinker, and T. Iskratsch, “The Role of the Dystrophin Glycoprotein Complex in Muscle Cell Mechano Transduction,” Communications Biology 5, no. 1 (2022): 1022.

[101] A. Starosta and P. Konieczny, “Therapeutic Aspects of Cell Signaling and Communication in Duchenne Muscular Dystrophy,” Cellular and Molecular Life Sciences 78, no. 11 (2021): 4867–4891.

[102] S. Chaouch, V. Mouly, A. Goyenvalle, et al., “Immortalized Skin Fibroblasts Expressing Conditional MyoD as a Renewable and Reliable Source of Converted human Muscle Cells to Assess Therapeutic Strategies for Muscular Dystrophies: Validation of an Exon-skipping Approach to Restore Dystrophin in Duchenne Muscular Dystrophy Cells,” Human Gene Therapy 20, no. 7 (2009): 784–790.

[103] K. Mamchaoui, C. Trollet, A. Bigot, et al., “Immortalized Pathological human Myoblasts: Towards a Universal Tool for the Study of Neuromuscular Disorders,” Skeletal Muscle 1 (2011): 34.

[104] M. Thorley, S. Duguez, E. M. C. Mazza, et al., “Skeletal Muscle Characteristics Are Preserved in hTERT/cdk4 human Myogenic Cell Lines,” Skeletal Muscle 6, no. 1 (2016): 43.

[105] S. Kimura, T. Noda, and T. Yoshimori, “Dissection of the Autophagosome Maturation Process by a Novel Reporter Protein, Tandem Fluorescent-tagged LC3,” Autophagy 3, no. 5 (2007): 452–460.

[106] S. Gao, C. Alarcon, G. Sapkota, et al., “Ubiquitin Ligase Nedd4L Targets Activated Smad2/3 to Limit TGF-beta Signaling,” Molecular Cell 36, no. 3 (2009): 457–468.

[107] C. Meyer, A. Garzia, P. Morozov, H. Molina, and T. Tuschl, “The G3BP1-Family-USP10 Deubiquitinase Complex Rescues Ubiquitinated 40S Subunits of Ribosomes Stalled in Translation From Lysosomal Degradation,” Molecular Cell 77, no. 6 (2020): 1193–1205.e5.

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