Engineering neuromuscular organoids from human induced pluripotent stem cells

Advancements in human induced pluripotent stem cell (hiPSC) biology, organoid technology and bioengineering approaches have recently opened new possibilities to mimic in vitro human tissues, organs and systems for basic science investigations, disease modeling and drug screening, and for the development of regenerative medicine strategies. Despite this multidisciplinary growing research field is holding promises, the integration of the multifaced scientific approaches is challenging, thus this remains a growing area of research. Here, we integrated biomaterials science and tissue engineering strategies together with hiPSC and organoid technologies to model and study the human neuromuscular (NM) system development and (dys)function. The NM system includes the skeletal muscle, and its stem cell compartment, functionally associated with a neural network able to evoke skeletal muscle contraction upon neuronal stimulation. To reach this aim, we used hydrogels, decellularized skeletal muscle or PDMS-based micropatterned substrates to derive self-assembled, tissue-engineered or patterned neuromuscular organoids (NMOs) from hiPSCs. Such organoid-based models were used to investigate the biological role of biomimetic materials during organoid production, but also to generate platforms in which mimic and study pathological features of genetically driven pathologies, such as Duchenne muscular dystrophy (DMD), or acquired conditions, as acute muscular damage and cancer-induced muscle cachexia. We demonstrated that the integration of biomimetic ECM materials and bioengineering-driven design principles with hiPSC-derived NMOs technology enables the generation of advanced in vitro platforms that mimic the human NM system, with improved reproducibility, functional relevance, and translational potential. These innovative platforms not only enhanced our understanding of NM development and function, but also provide valuable tools for studying pathological conditions and screening potential therapeutic interventions. The interdisciplinary approach presented here paves the way for future advancements in regenerative medicine and personalized healthcare, offering new possibilities for addressing NM disorders and, possibly, for improving in the future patient outcomes.