Advancements in 2D and 3D In Vitro Models for Studying Neuromuscular Diseases

Abstract

Neuromuscular diseases (NMDs) are a genetically or clinically heterogeneous group of diseases that involve injury or dysfunction of neuromuscular tissue components, including peripheral motor neurons, skeletal muscles, and neuromuscular junctions. To study NMDs and develop potential therapies, remarkable progress has been made in generating in vitro neuromuscular models using engineering approaches to recapitulate the complex physical and biochemical microenvironments of 3D human neuromuscular tissues. In this review, we discuss recent studies focusing on the development of in vitro co-culture models of human motor neurons and skeletal muscles, with the pros and cons of each approach. Furthermore, we explain how neuromuscular in vitro models recapitulate certain aspects of specific NMDs, including amyotrophic lateral sclerosis and muscular dystrophy. Research on neuromuscular organoids (NMO) will continue to co-develop to better mimic tissues in vivo and will provide a better understanding of the development of the neuromuscular tissue, mechanisms of NMD action, and tools applicable to preclinical studies, including drug screening and toxicity tests.

Aberrant evoked calcium signaling and nAChR cluster morphology in a SOD1 D90A hiPSC-derived neuromuscular model

“…Additionally, myogenic maturation features such as sarcomeric striation and spontaneous contraction have been described using sphere-based cultures of myogenic progenitors derived from hiPSC ( Hosoyama et al, 2014 ; Jiwlawat et al, 2017 ). Even though these methods significantly enhanced the general degree of maturation that myotubes can reach in vitro , achieving the formation of neuromuscular synapses similar in function and/or morphology to NMJs remained challenging (reviewed in Luttrell et al, 2021 ; Lynch et al, 2022 ; Gazzola and Martinat, 2023 ; Kim et al, 2023 ).…”

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“…Additionally, myogenic maturation features such as sarcomeric striation and spontaneous contraction have been described using sphere-based cultures of myogenic progenitors derived from hiPSC (Hosoyama et al, 2014;Jiwlawat et al, 2017). Even though these methods significantly enhanced the general degree of maturation that myotubes can reach in vitro, achieving the formation of neuromuscular synapses similar in function and/or morphology to NMJs remained challenging (reviewed in Luttrell et al, 2021;Lynch et al, 2022;Gazzola and Martinat, 2023;Kim et al, 2023).…”

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“…Additionally, myogenic maturation features such as sarcomeric striation and spontaneous contraction have been described using sphere-based cultures of myogenic progenitors derived from hiPSC (Hosoyama et al, 2014;Jiwlawat et al, 2017). Even though these methods significantly enhanced the general degree of maturation that myotubes can reach in vitro, achieving the formation of neuromuscular synapses similar in function and/or morphology to NMJs remained challenging (reviewed in Luttrell et al, 2021;Lynch et al, 2022;Gazzola and Martinat, 2023;Kim et al, 2023).…”