{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,1]],"date-time":"2026-09-01T15:44:31Z","timestamp":1788277471112,"version":"build-2803163510"},"reference-count":118,"publisher":"MDPI AG","issue":"20","license":[{"start":{"date-parts":[[2023,10,18]],"date-time":"2023-10-18T00:00:00Z","timestamp":1697587200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Association Fran\u00e7aise contre les Myopathies-T\u00e9l\u00e9thon","award":["ANR-21-CE14-0018-02"],"award-info":[{"award-number":["ANR-21-CE14-0018-02"]}]},{"name":"ANR CLASS","award":["ANR-21-CE14-0018-02"],"award-info":[{"award-number":["ANR-21-CE14-0018-02"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Over the past 20 years, the use of pluripotent stem cells to mimic the complexities of the human neuromuscular junction has received much attention. Deciphering the key mechanisms underlying the establishment and maturation of this complex synapse has been driven by the dual goals of addressing developmental questions and gaining insight into neuromuscular disorders. This review aims to summarise the evolution and sophistication of in vitro neuromuscular junction models developed from the first differentiation of human embryonic stem cells into motor neurons to recent neuromuscular organoids. We also discuss the potential offered by these models to decipher different neuromuscular diseases characterised by defects in the presynaptic compartment, the neuromuscular junction, and the postsynaptic compartment. Finally, we discuss the emerging field that considers the use of these techniques in drug screening assay and the challenges they will face in the future.<\/jats:p>","DOI":"10.3390\/ijms242015291","type":"journal-article","created":{"date-parts":[[2023,10,18]],"date-time":"2023-10-18T09:55:06Z","timestamp":1697622906000},"page":"15291","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Unlocking the Complexity of Neuromuscular Diseases: Insights from Human Pluripotent Stem Cell-Derived Neuromuscular Junctions"],"prefix":"10.3390","volume":"24","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-5231-099X","authenticated-orcid":false,"given":"Morgan","family":"Gazzola","sequence":"first","affiliation":[{"name":"INSERM U861, Institute for Stem Cell Therapy and Exploration of Monogenic Diseases, 91100 Corbeil-Essonnes, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5234-1064","authenticated-orcid":false,"given":"C\u00e9cile","family":"Martinat","sequence":"additional","affiliation":[{"name":"INSERM U861, Institute for Stem Cell Therapy and Exploration of Monogenic Diseases, 91100 Corbeil-Essonnes, France"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,10,18]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"73","DOI":"10.3233\/JND-140045","article-title":"The Epidemiology of Neuromuscular Disorders: A Comprehensive Overview of the Literature","volume":"2","author":"Deenen","year":"2015","journal-title":"J. Neuromuscul. Dis."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"76","DOI":"10.1016\/j.nmd.2022.12.002","article-title":"The 2023 version of the gene table of neuromuscular disorders (nuclear genome)","volume":"33","author":"Benarroch","year":"2023","journal-title":"Neuromuscul. Disord."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"827","DOI":"10.1111\/joa.13260","article-title":"Comparative anatomy of the mammalian neuromuscular junction","volume":"237","author":"Boehm","year":"2020","journal-title":"J. Anat."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1859","DOI":"10.1093\/nar\/gkab012","article-title":"Human pathways in animal models: Possibilities and limitations","volume":"49","author":"Doncheva","year":"2021","journal-title":"Nucleic Acids Res."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"26","DOI":"10.1186\/s13395-017-0143-9","article-title":"Are mice good models for human neuromuscular disease? Comparing muscle excursions in walking between mice and humans","volume":"7","author":"Hu","year":"2017","journal-title":"Skelet. Muscle"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"2348","DOI":"10.1016\/j.celrep.2017.11.008","article-title":"Cellular and Molecular Anatomy of the Human Neuromuscular Junction","volume":"21","author":"Jones","year":"2017","journal-title":"Cell Rep."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1461","DOI":"10.1172\/JCI128411","article-title":"Neuromuscular junctions are stable in patients with cancer cachexia","volume":"130","author":"Boehm","year":"2020","journal-title":"J. Clin. Investig."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"709","DOI":"10.1016\/j.tins.2020.07.002","article-title":"Harmony Lost: Cell\u2013Cell Communication at the Neuromuscular Junction in Motor Neuron Disease","volume":"43","author":"Gromova","year":"2020","journal-title":"Trends Neurosci."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Slater, C. (2017). The Structure of Human Neuromuscular Junctions: Some Unanswered Molecular Questions. Int. J. Mol. Sci., 18.","DOI":"10.3390\/ijms18102183"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"dev193920","DOI":"10.1242\/dev.193920","article-title":"Building neuromuscular junctions in vitro","volume":"147","author":"Barbeau","year":"2020","journal-title":"Development"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"2211059","DOI":"10.1002\/adma.202211059","article-title":"Advances in In Vitro Models of Neuromuscular Junction: Focusing on Organ-on-a-Chip, Organoids, and Biohybrid Robotics","volume":"35","author":"Leng","year":"2023","journal-title":"Adv. Mater."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"215","DOI":"10.1038\/nbt1063","article-title":"Specification of motoneurons from human embryonic stem cells","volume":"23","author":"Li","year":"2005","journal-title":"Nat. Biotechnol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"385","DOI":"10.1016\/S0092-8674(02)00835-8","article-title":"Directed Differentiation of Embryonic Stem Cells into Motor Neurons","volume":"110","author":"Wichterle","year":"2002","journal-title":"Cell"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"224","DOI":"10.1016\/j.expneurol.2005.06.021","article-title":"Enhancer-specified GFP-based FACS purification of human spinal motor neurons from embryonic stem cells","volume":"196","author":"Nakano","year":"2005","journal-title":"Exp. Neurol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"561","DOI":"10.1016\/j.stemcr.2015.02.010","article-title":"Modeling the Early Phenotype at the Neuromuscular Junction of Spinal Muscular Atrophy Using Patient-Derived iPSCs","volume":"4","author":"Yoshida","year":"2015","journal-title":"Stem Cell Rep."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"115","DOI":"10.1016\/j.celrep.2015.12.018","article-title":"Direct Lineage Reprogramming Reveals Disease-Specific Phenotypes of Motor Neurons from Human ALS Patients","volume":"14","author":"Liu","year":"2016","journal-title":"Cell Rep."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1186\/s13619-023-00159-6","article-title":"Generation of functional posterior spinal motor neurons from hPSCs-derived human spinal cord neural progenitor cells","volume":"12","author":"Xu","year":"2023","journal-title":"Cell Regen."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"434","DOI":"10.1016\/j.stem.2011.02.004","article-title":"Mutant Human Embryonic Stem Cells Reveal Neurite and Synapse Formation Defects in Type 1 Myotonic Dystrophy","volume":"8","author":"Marteyn","year":"2011","journal-title":"Cell Stem Cell"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1347","DOI":"10.1089\/ten.tec.2010.0040","article-title":"Neuromuscular Junction Formation Between Human Stem-Cell-Derived Motoneurons and Rat Skeletal Muscle in a Defined System","volume":"16","author":"Guo","year":"2010","journal-title":"Tissue Eng. Part C Methods"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"9602","DOI":"10.1016\/j.biomaterials.2011.09.014","article-title":"Neuromuscular junction formation between human stem cell-derived motoneurons and human skeletal muscle in a defined system","volume":"32","author":"Guo","year":"2011","journal-title":"Biomaterials"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"663","DOI":"10.1016\/j.cell.2006.07.024","article-title":"Induction of Pluripotent Stem Cells from Mouse Embryonic and Adult Fibroblast Cultures by Defined Factors","volume":"126","author":"Takahashi","year":"2006","journal-title":"Cell"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"475","DOI":"10.1007\/s12015-011-9329-4","article-title":"Developmental and Functional Nature of Human iPSC Derived Motoneurons","volume":"9","author":"Stockmann","year":"2013","journal-title":"Stem Cell Rev. Rep."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1291","DOI":"10.1523\/JNEUROSCI.2126-14.2015","article-title":"Motoneurons Derived from Induced Pluripotent Stem Cells Develop Mature Phenotypes Typical of Endogenous Spinal Motoneurons","volume":"35","author":"Toma","year":"2015","journal-title":"J. Neurosci."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"982","DOI":"10.1002\/dneu.20953","article-title":"Role of extracellular matrix proteins and their receptors in the development of the vertebrate neuromuscular junction","volume":"71","author":"Singhal","year":"2011","journal-title":"Dev. Neurobiol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"473","DOI":"10.3389\/fncel.2015.00473","article-title":"Generation of Functional Neuromuscular Junctions from Human Pluripotent Stem Cell Lines","volume":"9","author":"Puttonen","year":"2015","journal-title":"Front. Cell. Neurosci."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"328","DOI":"10.1016\/j.scr.2015.07.005","article-title":"Formation and characterisation of neuromuscular junctions between hiPSC derived motoneurons and myotubes","volume":"15","author":"Demestre","year":"2015","journal-title":"Stem Cell Res."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"e124299","DOI":"10.1172\/jci.insight.124299","article-title":"iPSC-derived functional human neuromuscular junctions model the pathophysiology of neuromuscular diseases","volume":"4","author":"Lin","year":"2019","journal-title":"JCI Insight"},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Mazaleyrat, K., Badja, C., Broucqsault, N., Chevalier, R., Laberthonni\u00e8re, C., Dion, C., Baldasseroni, L., El-Yazidi, C., Thomas, M., and Bachelier, R. (2020). Multilineage Differentiation for Formation of Innervated Skeletal Muscle Fibers from Healthy and Diseased Human Pluripotent Stem Cells. Cells, 9.","DOI":"10.3390\/cells9061531"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"605","DOI":"10.1007\/s12195-020-00624-1","article-title":"A Chemically Defined Common Medium for Culture of C2C12 Skeletal Muscle and Human Induced Pluripotent Stem Cell Derived Spinal Spheroids","volume":"13","author":"Besser","year":"2020","journal-title":"Cell. Mol. Bioeng."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"661","DOI":"10.1089\/ten.tea.2021.0202","article-title":"Contraction Control of Aligned Myofiber Sheet Tissue by Parallel Oriented Induced Pluripotent Stem Cell-Derived Neurons","volume":"28","author":"Takahashi","year":"2022","journal-title":"Tissue Eng. Part A"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"389","DOI":"10.1146\/annurev.neuro.22.1.389","article-title":"Development of the Vertebrate Neuromuscular Junction","volume":"22","author":"Sanes","year":"1999","journal-title":"Annu. Rev. Neurosci."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"3901","DOI":"10.1242\/jcs.031047","article-title":"Identity, developmental restriction and reactivity of extralaminar cells capping mammalian neuromuscular junctions","volume":"121","author":"Court","year":"2008","journal-title":"J. Cell Sci."},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"H\u00f6rner, S.J., Couturier, N., Bruch, R., Koch, P., Hafner, M., and Rudolf, R. (2021). hiPSC-Derived Schwann Cells Influence Myogenic Differentiation in Neuromuscular Cocultures. Cells, 10.","DOI":"10.3390\/cells10123292"},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Umbach, J.A., Adams, K.L., Gundersen, C.B., and Novitch, B.G. (2012). Functional Neuromuscular Junctions Formed by Embryonic Stem Cell-Derived Motor Neurons. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0036049"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"134","DOI":"10.1016\/j.stem.2015.10.002","article-title":"Functional Connectivity under Optogenetic Control Allows Modeling of Human Neuromuscular Disease","volume":"18","author":"Steinbeck","year":"2016","journal-title":"Cell Stem Cell"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"046031","DOI":"10.1088\/1741-2560\/8\/4\/046031","article-title":"Propagation of action potential activity in a predefined microtunnel neural network","volume":"8","author":"Pan","year":"2011","journal-title":"J. Neural Eng."},{"key":"ref_37","unstructured":"Park, H.S., Liu, S., McDonald, J., Thakor, N., and Yang, I.H. (2013, January 3\u20137). Neuromuscular junction in a microfluidic device. Proceedings of the 2013 35th Annual International Conference of the IEEE Engineering in Medicine and Biology Society (EMBC), Osaka, Japan."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"e1501429","DOI":"10.1126\/sciadv.1501429","article-title":"Microfluidic device for the formation of optically excitable, three-dimensional, compartmentalized motor units","volume":"2","author":"Uzel","year":"2016","journal-title":"Sci. Adv."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"64","DOI":"10.1016\/j.biomaterials.2018.02.047","article-title":"Stem cell derived phenotypic human neuromuscular junction model for dose response evaluation of therapeutics","volume":"166","author":"Santhanam","year":"2018","journal-title":"Biomaterials"},{"key":"ref_40","doi-asserted-by":"crossref","unstructured":"Bellmann, J., Goswami, R.Y., Girardo, S., Rein, N., Hosseinzadeh, Z., Hicks, M.R., Busskamp, V., Pyle, A.D., Werner, C., and Sterneckert, J. (2019). A customizable microfluidic platform for medium-throughput modeling of neuromuscular circuits. Biomaterials, 225.","DOI":"10.1016\/j.biomaterials.2019.119537"},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Machado, C.B., Pluchon, P., Harley, P., Rigby, M., Sabater, V.G., Stevenson, D.C., Hynes, S., Lowe, A., Burrone, J., and Viasnoff, V. (2019). In Vitro Modeling of Nerve\u2013Muscle Connectivity in a Compartmentalized Tissue Culture Device. Adv. Biosyst., 3.","DOI":"10.1002\/adbi.201800307"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"19841","DOI":"10.1073\/pnas.1907051116","article-title":"Neuromuscular actuation of biohybrid motile bots","volume":"116","author":"Aydin","year":"2019","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"2213","DOI":"10.1016\/j.stemcr.2021.03.029","article-title":"Human motor units in microfluidic devices are impaired by FUS mutations and improved by HDAC6 inhibition","volume":"16","author":"Krasnow","year":"2021","journal-title":"Stem Cell Rep."},{"key":"ref_44","first-page":"62959","article-title":"Generation of Human Motor Units with Functional Neuromuscular Junctions in Microfluidic Devices","volume":"175","author":"Krasnow","year":"2021","journal-title":"J. Vis. Exp."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"5","DOI":"10.1186\/s13024-022-00591-3","article-title":"FUS-ALS hiPSC-derived astrocytes impair human motor units through both gain-of-toxicity and loss-of-support mechanisms","volume":"18","author":"Terrie","year":"2023","journal-title":"Mol. Neurodegener."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"e44530","DOI":"10.7554\/eLife.44530","article-title":"A 3D culture model of innervated human skeletal muscle enables studies of the adult neuromuscular junction","volume":"8","author":"Lippmann","year":"2019","journal-title":"eLife"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"486","DOI":"10.1016\/j.jbiosc.2019.10.004","article-title":"Novel neuromuscular junction model in 2D and 3D myotubes co-cultured with induced pluripotent stem cell-derived motor neurons","volume":"129","author":"Yoshioka","year":"2020","journal-title":"J. Biosci. Bioeng."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"1232","DOI":"10.7150\/thno.25735","article-title":"Quantification of human neuromuscular function through optogenetics","volume":"9","author":"Vila","year":"2019","journal-title":"Theranostics"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"eaat5847","DOI":"10.1126\/sciadv.aat5847","article-title":"Microphysiological 3D model of amyotrophic lateral sclerosis (ALS) from human iPS-derived muscle cells and optogenetic motor neurons","volume":"4","author":"Osaki","year":"2018","journal-title":"Sci. Adv."},{"key":"ref_50","doi-asserted-by":"crossref","unstructured":"Harley, P., Paredes-Redondo, A., Grenci, G., Viasnoff, V., Lin, Y.-Y., and Lieberam, I. (2023). 3D Compartmentalised Human Pluripotent Stem Cell\u2013derived Neuromuscular Co-cultures. Bio-Protocol, 13.","DOI":"10.21769\/BioProtoc.4624"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"346","DOI":"10.1089\/ten.tec.2018.0062","article-title":"Bioinspired Three-Dimensional Human Neuromuscular Junction Development in Suspended Hydrogel Arrays","volume":"24","author":"Dixon","year":"2018","journal-title":"Tissue Eng. Part C Methods"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"1025","DOI":"10.1038\/s41467-020-14930-9","article-title":"Neural cell integration into 3D bioprinted skeletal muscle constructs accelerates restoration of muscle function","volume":"11","author":"Kim","year":"2020","journal-title":"Nat. Commun."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"2100581","DOI":"10.1002\/adhm.202100581","article-title":"Promoting Long-Term Cultivation of Motor Neurons for 3D Neuromuscular Junction Formation of 3D In Vitro Using Central-Nervous-Tissue-Derived Bioink","volume":"10","author":"Kong","year":"2021","journal-title":"Adv. Healthc. Mater."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"18","DOI":"10.1038\/s41536-023-00292-5","article-title":"3D bioprinting using a new photo-crosslinking method for muscle tissue restoration","volume":"8","author":"Lee","year":"2023","journal-title":"NPJ Regen. Med."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"571","DOI":"10.1016\/j.stem.2023.04.011","article-title":"Human pluripotent-stem-cell-derived organoids for drug discovery and evaluation","volume":"30","author":"Vandana","year":"2023","journal-title":"Cell Stem Cell"},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"172","DOI":"10.1016\/j.stem.2019.12.007","article-title":"Self-Organizing 3D Human Trunk Neuromuscular Organoids","volume":"26","author":"Fischer","year":"2020","journal-title":"Cell Stem Cell"},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"441","DOI":"10.1016\/j.tins.2012.04.005","article-title":"Molecular mechanisms underlying maturation and maintenance of the vertebrate neuromuscular junction","volume":"35","author":"Shi","year":"2012","journal-title":"Trends Neurosci."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"1913","DOI":"10.1016\/j.cell.2020.11.017","article-title":"Generation of Functional Human 3D Cortico-Motor Assembloids","volume":"183","author":"Andersen","year":"2020","journal-title":"Cell"},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"4744","DOI":"10.1038\/s41467-021-24776-4","article-title":"Human sensorimotor organoids derived from healthy and amyotrophic lateral sclerosis stem cells form neuromuscular junctions","volume":"12","author":"Pereira","year":"2021","journal-title":"Nat. Commun."},{"key":"ref_60","doi-asserted-by":"crossref","unstructured":"Shin, M.-K., Bang, J.S., Lee, J.E., Tran, H.-D., Park, G., Lee, D.R., and Jo, J. (2022). Generation of Skeletal Muscle Organoids from Human Pluripotent Stem Cells to Model Myogenesis and Muscle Regeneration. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms23095108"},{"key":"ref_61","doi-asserted-by":"crossref","unstructured":"Straka, T., Vita, V., Prokshi, K., H\u00f6rner, S.J., Khan, M.M., Pirazzini, M., Williams, M.P.I., Hafner, M., Zaglia, T., and Rudolf, R. (2018). Postnatal Development and Distribution of Sympathetic Innervation in Mouse Skeletal Muscle. Int. J. Mol. Sci., 19.","DOI":"10.3390\/ijms19071935"},{"key":"ref_62","doi-asserted-by":"crossref","unstructured":"van den Bos, M.A.J., Geevasinga, N., Higashihara, M., Menon, P., and Vucic, S. (2019). Pathophysiology and Diagnosis of ALS: Insights from Advances in Neurophysiological Techniques. Int. J. Mol. Sci., 20.","DOI":"10.3390\/ijms20112818"},{"key":"ref_63","doi-asserted-by":"crossref","unstructured":"Cappello, V., and Francolini, M. (2017). Neuromuscular Junction Dismantling in Amyotrophic Lateral Sclerosis. Int. J. Mol. Sci., 18.","DOI":"10.3390\/ijms18102092"},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"35","DOI":"10.1016\/j.jchemneu.2016.03.003","article-title":"Axonal degeneration, distal collateral branching and neuromuscular junction architecture alterations occur prior to symptom onset in the SOD1G93A mouse model of amyotrophic lateral sclerosis","volume":"76","author":"Clark","year":"2016","journal-title":"J. Chem. Neuroanat."},{"key":"ref_65","doi-asserted-by":"crossref","first-page":"131","DOI":"10.3389\/fncel.2014.00131","article-title":"The role of oxidative stress in degeneration of the neuromuscular junction in amyotrophic lateral sclerosis","volume":"8","author":"Pollari","year":"2014","journal-title":"Front. Cell. Neurosci."},{"key":"ref_66","doi-asserted-by":"crossref","first-page":"12","DOI":"10.1016\/j.mcn.2012.07.003","article-title":"Analysis of neuromuscular junctions and effects of anabolic steroid administration in the SOD1G93A mouse model of ALS","volume":"51","author":"Cappello","year":"2012","journal-title":"Mol. Cell. Neurosci."},{"key":"ref_67","doi-asserted-by":"crossref","first-page":"24515","DOI":"10.1038\/srep24515","article-title":"INaP selective inhibition reverts precocious inter- and motorneurons hyperexcitability in the Sod1-G93R zebrafish ALS model","volume":"6","author":"Benedetti","year":"2016","journal-title":"Sci. Rep."},{"key":"ref_68","doi-asserted-by":"crossref","first-page":"431","DOI":"10.3109\/21678421.2015.1055275","article-title":"Human TDP-43 and FUS selectively affect motor neuron maturation and survival in a murine cell model of ALS by non-cell-autonomous mechanisms","volume":"16","author":"Storch","year":"2015","journal-title":"Amyotroph. Lateral Scler. Front. Degener."},{"key":"ref_69","doi-asserted-by":"crossref","first-page":"1793","DOI":"10.1038\/s41593-019-0498-9","article-title":"FUS-mediated regulation of acetylcholine receptor transcription at neuromuscular junctions is compromised in amyotrophic lateral sclerosis","volume":"22","author":"Picchiarelli","year":"2019","journal-title":"Nat. Neurosci."},{"key":"ref_70","doi-asserted-by":"crossref","first-page":"2000133","DOI":"10.1002\/adtp.202000133","article-title":"A Human-Based Functional NMJ System for Personalized ALS Modeling and Drug Testing","volume":"3","author":"Guo","year":"2020","journal-title":"Adv. Ther."},{"key":"ref_71","doi-asserted-by":"crossref","unstructured":"Ting, H.-C., Su, H.-L., Chen, M.-F., Harn, H.-J., Lin, S.-Z., Chiou, T.-W., and Chang, C.-Y. (2022). Robust Generation of Ready-to-Use Cryopreserved Motor Neurons from Human Pluripotent Stem Cells for Disease Modeling. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms232113462"},{"key":"ref_72","doi-asserted-by":"crossref","unstructured":"Badu-Mensah, A., Guo, X., Nimbalkar, S., Cai, Y., and Hickman, J.J. (2022). ALS mutations in both human skeletal muscle and motoneurons differentially affects neuromuscular junction integrity and function. Biomaterials, 289.","DOI":"10.1016\/j.biomaterials.2022.121752"},{"key":"ref_73","doi-asserted-by":"crossref","unstructured":"Massih, B., Veh, A., Schenke, M., Mungwa, S., Seeger, B., Selvaraj, B.T., Chandran, S., Reinhardt, P., Sterneckert, J., and Hermann, A. (2023). A 3D cell culture system for bioengineering human neuromuscular junctions to model ALS. Front. Cell Dev. Biol., 11.","DOI":"10.3389\/fcell.2023.996952"},{"key":"ref_74","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1101\/gr.275939.121","article-title":"Meta-analysis of human and mouse ALS astrocytes reveals multi-omic signatures of inflammatory reactive states","volume":"32","author":"Ziff","year":"2022","journal-title":"Genome Res."},{"key":"ref_75","doi-asserted-by":"crossref","first-page":"481","DOI":"10.1093\/brain\/awab328","article-title":"Astrocytes display cell autonomous and diverse early reactive states in familial amyotrophic lateral sclerosis","volume":"145","author":"Taha","year":"2022","journal-title":"Brain"},{"key":"ref_76","doi-asserted-by":"crossref","first-page":"131","DOI":"10.1055\/s-2001-15260","article-title":"Amyotrophic Lateral Sclerosis: Pathogenesis","volume":"21","author":"Brown","year":"2001","journal-title":"Semin. Neurol."},{"key":"ref_77","doi-asserted-by":"crossref","first-page":"52","DOI":"10.1038\/s41572-022-00380-8","article-title":"Spinal muscular atrophy","volume":"8","author":"Mercuri","year":"2022","journal-title":"Nat. Rev. Dis. Primer"},{"key":"ref_78","doi-asserted-by":"crossref","first-page":"11696","DOI":"10.1038\/srep11696","article-title":"Decay in survival motor neuron and plastin 3 levels during differentiation of iPSC-derived human motor neurons","volume":"5","author":"Bernal","year":"2015","journal-title":"Sci. Rep."},{"key":"ref_79","doi-asserted-by":"crossref","first-page":"165ra162","DOI":"10.1126\/scitranslmed.3004108","article-title":"Genetic Correction of Human Induced Pluripotent Stem Cells from Patients with Spinal Muscular Atrophy","volume":"4","author":"Corti","year":"2012","journal-title":"Sci. Transl. Med."},{"key":"ref_80","doi-asserted-by":"crossref","first-page":"276","DOI":"10.1093\/brain\/awy330","article-title":"Key role of SMN\/SYNCRIP and RNA-Motif 7 in spinal muscular atrophy: RNA-Seq and motif analysis of human motor neurons","volume":"142","author":"Rizzo","year":"2019","journal-title":"Brain"},{"key":"ref_81","doi-asserted-by":"crossref","first-page":"441","DOI":"10.1007\/s00018-022-04450-8","article-title":"Moxifloxacin rescues SMA phenotypes in patient-derived cells and animal model","volume":"79","author":"Januel","year":"2022","journal-title":"Cell. Mol. Life Sci."},{"key":"ref_82","doi-asserted-by":"crossref","first-page":"89","DOI":"10.1038\/nbt.3049","article-title":"Combinatorial analysis of developmental cues efficiently converts human pluripotent stem cells into multiple neuronal subtypes","volume":"33","author":"Maury","year":"2015","journal-title":"Nat. Biotechnol."},{"key":"ref_83","doi-asserted-by":"crossref","first-page":"1110499","DOI":"10.3389\/fimmu.2023.1110499","article-title":"Inflammation and autoimmune myasthenia gravis","volume":"14","author":"Huda","year":"2023","journal-title":"Front. Immunol."},{"key":"ref_84","doi-asserted-by":"crossref","first-page":"57","DOI":"10.1186\/s13023-019-1025-5","article-title":"Congenital myasthenic syndromes","volume":"14","author":"Finsterer","year":"2019","journal-title":"Orphanet J. Rare Dis."},{"key":"ref_85","doi-asserted-by":"crossref","first-page":"707","DOI":"10.1007\/s00401-022-02475-8","article-title":"Severe congenital myasthenic syndromes caused by agrin mutations affecting secretion by motoneurons","volume":"144","author":"Jacquier","year":"2022","journal-title":"Acta Neuropathol."},{"key":"ref_86","doi-asserted-by":"crossref","unstructured":"Smith, V.M., Nguyen, H., Rumsey, J.W., Long, C.J., Shuler, M.L., and Hickman, J.J. (2021). A Functional Human-on-a-Chip Autoimmune Disease Model of Myasthenia Gravis for Development of Therapeutics. Front. Cell Dev. Biol., 9.","DOI":"10.3389\/fcell.2021.745897"},{"key":"ref_87","doi-asserted-by":"crossref","first-page":"572","DOI":"10.1016\/j.neurol.2016.08.003","article-title":"Unravelling the myotonic dystrophy type 1 clinical spectrum: A systematic registry-based study with implications for disease classification","volume":"172","author":"Dogan","year":"2016","journal-title":"Rev. Neurol."},{"key":"ref_88","doi-asserted-by":"crossref","first-page":"2766","DOI":"10.1093\/nar\/29.13.2766","article-title":"In vivo co-localisation of MBNL protein with DMPK expanded-repeat transcripts","volume":"29","author":"Fardaei","year":"2001","journal-title":"Nucleic Acids Res."},{"key":"ref_89","doi-asserted-by":"crossref","unstructured":"Xing, X., Kumari, A., Brown, J., and Brook, J.D. (2021). Disrupting the Molecular Pathway in Myotonic Dystrophy. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms222413225"},{"key":"ref_90","doi-asserted-by":"crossref","unstructured":"Tahraoui-Bories, J., M\u00e9rien, A., Gonz\u00e1lez-Barriga, A., Lain\u00e9, J., Leteur, C., Polv\u00e8che, H., Carteron, A., De Lamotte, J.D., Nicoleau, C., and Polentes, J. (2023). MBNL-dependent impaired development within the neuromuscular system in myotonic dystrophy type 1. Neuropathol. Appl. Neurobiol., 49.","DOI":"10.1111\/nan.12876"},{"key":"ref_91","doi-asserted-by":"crossref","first-page":"1183101","DOI":"10.3389\/fphys.2023.1183101","article-title":"Duchenne muscular dystrophy: Disease mechanism and therapeutic strategies","volume":"14","author":"Hosny","year":"2023","journal-title":"Front. Physiol."},{"key":"ref_92","doi-asserted-by":"crossref","first-page":"eabi8787","DOI":"10.1126\/sciadv.abi8787","article-title":"Optogenetic modeling of human neuromuscular circuits in Duchenne muscular dystrophy with CRISPR and pharmacological corrections","volume":"7","author":"Harley","year":"2021","journal-title":"Sci. Adv."},{"key":"ref_93","doi-asserted-by":"crossref","first-page":"585","DOI":"10.1056\/NEJM199403033300901","article-title":"A Controlled Trial of Riluzole in Amyotrophic Lateral Sclerosis","volume":"330","author":"Bensimon","year":"1994","journal-title":"N. Engl. J. Med."},{"key":"ref_94","doi-asserted-by":"crossref","first-page":"505","DOI":"10.1016\/S1474-4422(17)30115-1","article-title":"Safety and efficacy of edaravone in well defined patients with amyotrophic lateral sclerosis: A randomised, double-blind, placebo-controlled trial","volume":"16","author":"Abe","year":"2017","journal-title":"Lancet Neurol."},{"key":"ref_95","doi-asserted-by":"crossref","first-page":"919","DOI":"10.1056\/NEJMoa1916945","article-title":"Trial of Sodium Phenylbutyrate\u2013Taurursodiol for Amyotrophic Lateral Sclerosis","volume":"383","author":"Paganoni","year":"2020","journal-title":"N. Engl. J. Med."},{"key":"ref_96","doi-asserted-by":"crossref","first-page":"1713","DOI":"10.1056\/NEJMoa1706198","article-title":"Single-Dose Gene-Replacement Therapy for Spinal Muscular Atrophy","volume":"377","author":"Mendell","year":"2017","journal-title":"N. Engl. J. Med."},{"key":"ref_97","doi-asserted-by":"crossref","first-page":"46","DOI":"10.1124\/jpet.113.212407","article-title":"Pharmacology of a Central Nervous System Delivered 2\u2032-O-Methoxyethyl\u2013Modified Survival of Motor Neuron Splicing Oligonucleotide in Mice and Nonhuman Primates","volume":"350","author":"Rigo","year":"2014","journal-title":"J. Pharmacol. Exp. Ther."},{"key":"ref_98","doi-asserted-by":"crossref","first-page":"6501","DOI":"10.1021\/acs.jmedchem.8b00741","article-title":"Discovery of Risdiplam, a Selective Survival of Motor Neuron-2 (SMN2) Gene Splicing Modifier for the Treatment of Spinal Muscular Atrophy (SMA)","volume":"61","author":"Ratni","year":"2018","journal-title":"J. Med. Chem."},{"key":"ref_99","doi-asserted-by":"crossref","first-page":"160","DOI":"10.1016\/j.jocn.2019.02.004","article-title":"Effectiveness and safety of tacrolimus therapy for myasthenia gravis: A single arm meta-analysis","volume":"63","author":"Wang","year":"2019","journal-title":"J. Clin. Neurosci."},{"key":"ref_100","doi-asserted-by":"crossref","first-page":"494","DOI":"10.1196\/annals.1254.064","article-title":"Mycophenolate Mofetil for Myasthenia Gravis A Double-Blind, Placebo-Controlled Pilot Study","volume":"998","author":"Meriggioli","year":"2003","journal-title":"Ann. N. Y. Acad. Sci."},{"key":"ref_101","doi-asserted-by":"crossref","first-page":"448","DOI":"10.1212\/WNL.55.3.448","article-title":"Retrospective analysis of the use of cyclosporine in myasthenia gravis","volume":"55","author":"Ciafaloni","year":"2000","journal-title":"Neurology"},{"key":"ref_102","doi-asserted-by":"crossref","first-page":"57","DOI":"10.1212\/WNL.0000000000002795","article-title":"A randomized controlled trial of methotrexate for patients with generalized myasthenia gravis","volume":"87","author":"Pasnoor","year":"2016","journal-title":"Neurology"},{"key":"ref_103","doi-asserted-by":"crossref","first-page":"31","DOI":"10.1002\/mus.10133","article-title":"Use of intravenous pulsed cyclophosphamide in severe, generalized myasthenia gravis","volume":"26","author":"Schottlender","year":"2002","journal-title":"Muscle Nerve"},{"key":"ref_104","doi-asserted-by":"crossref","first-page":"259","DOI":"10.1177\/1756285611411503","article-title":"Response of patients with refractory myasthenia gravis to rituximab: A retrospective study","volume":"4","author":"Nowak","year":"2011","journal-title":"Ther. Adv. Neurol. Disord."},{"key":"ref_105","doi-asserted-by":"crossref","first-page":"1327","DOI":"10.1002\/acn3.51121","article-title":"Consistent improvement with eculizumab across muscle groups in myasthenia gravis","volume":"7","author":"Mantegazza","year":"2020","journal-title":"Ann. Clin. Transl. Neurol."},{"key":"ref_106","doi-asserted-by":"crossref","first-page":"526","DOI":"10.1016\/S1474-4422(21)00159-9","article-title":"Safety, efficacy, and tolerability of efgartigimod in patients with generalised myasthenia gravis (ADAPT): A multicentre, randomised, placebo-controlled, phase 3 trial","volume":"20","author":"Howard","year":"2021","journal-title":"Lancet Neurol."},{"key":"ref_107","doi-asserted-by":"crossref","first-page":"875","DOI":"10.1007\/s40265-021-01512-2","article-title":"Casimersen: First Approval","volume":"81","author":"Shirley","year":"2021","journal-title":"Drugs"},{"key":"ref_108","doi-asserted-by":"crossref","first-page":"1027","DOI":"10.1007\/s40265-020-01339-3","article-title":"Viltolarsen: First Approval","volume":"80","author":"Dhillon","year":"2020","journal-title":"Drugs"},{"key":"ref_109","doi-asserted-by":"crossref","first-page":"329","DOI":"10.1007\/s40265-020-01267-2","article-title":"Golodirsen: First Approval","volume":"80","author":"Heo","year":"2020","journal-title":"Drugs"},{"key":"ref_110","doi-asserted-by":"crossref","first-page":"45","DOI":"10.1067\/mpd.2001.109601","article-title":"Deflazacort treatment of Duchenne muscular dystrophy","volume":"138","author":"Biggar","year":"2001","journal-title":"J. Pediatr."},{"key":"ref_111","doi-asserted-by":"crossref","first-page":"1699","DOI":"10.1007\/s40265-016-0657-1","article-title":"Eteplirsen: First Global Approval","volume":"76","author":"Syed","year":"2016","journal-title":"Drugs"},{"key":"ref_112","doi-asserted-by":"crossref","unstructured":"Zaidman, C.M., Proud, C.M., McDonald, C.M., Lehman, K.J., Goedeker, N.L., Mason, S., Murphy, A.P., Guridi, M., Wang, S., and Reid, C. (2023). Delandistrogene Moxeparvovec Gene Therapy in Ambulatory Patients (Aged \u22654 to <8 Years) with Duchenne Muscular Dystrophy: 1-Year Interim Results from Study SRP-9001-103 (ENDEAVOR). Ann. Neurol., ana.26755.","DOI":"10.1002\/ana.26755"},{"key":"ref_113","doi-asserted-by":"crossref","unstructured":"Chipman, P.H., Zhang, Y., and Rafuse, V.F. (2014). A Stem-Cell Based Bioassay to Critically Assess the Pathology of Dysfunctional Neuromuscular Junctions. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0091643"},{"key":"ref_114","doi-asserted-by":"crossref","first-page":"421","DOI":"10.1038\/s41596-019-0248-1","article-title":"On-chip 3D neuromuscular model for drug screening and precision medicine in neuromuscular disease","volume":"15","author":"Osaki","year":"2020","journal-title":"Nat. Protoc."},{"key":"ref_115","doi-asserted-by":"crossref","first-page":"599","DOI":"10.1186\/s13287-021-02665-3","article-title":"Optogenetically controlled human functional motor endplate for testing botulinum neurotoxins","volume":"12","author":"Polentes","year":"2021","journal-title":"Stem Cell Res. Ther."},{"key":"ref_116","doi-asserted-by":"crossref","first-page":"eaaf3962","DOI":"10.1126\/scitranslmed.aaf3962","article-title":"The Src\/c-Abl pathway is a potential therapeutic target in amyotrophic lateral sclerosis","volume":"9","author":"Imamura","year":"2017","journal-title":"Sci. Transl. Med."},{"key":"ref_117","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.celrep.2014.03.019","article-title":"Intrinsic Membrane Hyperexcitability of Amyotrophic Lateral Sclerosis Patient-Derived Motor Neurons","volume":"7","author":"Wainger","year":"2014","journal-title":"Cell Rep."},{"key":"ref_118","doi-asserted-by":"crossref","first-page":"2855","DOI":"10.1093\/brain\/awy231","article-title":"Improved mobility with metformin in patients with myotonic dystrophy type 1: A randomized controlled trial","volume":"141","author":"Bassez","year":"2018","journal-title":"Brain"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/24\/20\/15291\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T21:08:51Z","timestamp":1760130531000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/24\/20\/15291"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,10,18]]},"references-count":118,"journal-issue":{"issue":"20","published-online":{"date-parts":[[2023,10]]}},"alternative-id":["ijms242015291"],"URL":"https:\/\/doi.org\/10.3390\/ijms242015291","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,10,18]]}}}