Magnetically induced nano-forces in neuromuscular organoids for disease modeling

*Neuromuscular disorders, whose incidence and prevalence have increased over time have currently no cure, and understanding their pathological mechanisms remains challenging due to the lack of reliable human in vitro models. A groundbreaking approach involves human 3D trunk neuromuscular organoids (NMOs) that replicate the cellular diversity and the functional dynamics of the neuromuscular system. However, the absence of in vivo-like external stimulations leads to reduced contractions and prevent terminal maturation, restraining their applicability for studying long-term neuromuscular diseases. The primary goal of this project is to design innovative multi-disciplinary approaches to mimic in vivo mechanical and brain inputs and provide new paradigms for organoid generation. A new method will be developed to apply spatiotemporally controlled nano-forces within NMOs, with the aim of enhancing the maturation of both the neural and muscle tissue. Additionally, magnetically induced nano-forces will be exploited to guide the reconstruction of the corticospinal tract within newly generated cerebral-NMOs and integrate brain inputs. Cutting-edge imaging, optogenetics, and single-nucleus RNA sequencing will be used for in-depth characterization of the outcomes. The hypothesis is that these stimuli will significantly promote NMOs maturation, resulting in a more sophisticated model for neuromuscular disease research. As case-study, the enhanced NMOs will be applied to model and explore the complex pathological mechanisms of X-linked spinal and bulbar muscular atrophy (SBMA). Patient-derived nano-forced cerebral-NMOs will be generated to investigate the impact of SBMA on neuromuscular system and corticospinal tract. Supporting this project will enable the implementation of innovative methodologies for the optimization of a relevant human in vitro system, with the potential to revolutionize research and treatment approaches for neuromuscular diseases.