14 | Extracellular matrix guides human neuromuscular organoid morphogenesis

Beatrice Auletta^1|2|3, P. Chiolerio^1|2, G. Cecconi^1|2, L. Sartore^1|2, L. Rossi^1|2, E. Maghin⁴, S. Angiolillo^3|5, S. Calabrò¹, O. Gagliano^3|5, C. Laterza^3|5, M. Cescon¹, Y. Torrente^6|7, C. Luni⁸, M. Piccoli⁴, N. Elvassore^3|5, A. Urciuolo^1|2 | ¹Department of Molecular Medicine, University of Padova, Padova, Italy; ²Neuromuscular Engineering lab, Istituto di Ricerca Pediatrica, Città della Speranza, Padova, Italy; ³Department of Industrial Engineering, University of Padova, Padova, Italy; ⁴Tissue Engineering lab, Istituto di Ricerca Pediatrica, Città della Speranza, Padova, Italy; ⁵Veneto Institute of Molecular Medicine, Padova, Italy; ⁶Stem Cell Laboratory, Dino Ferrari Center, Department of Pathophysiology and Transplantation, University of Milan, Milan, Italy; ⁷Neurology Unit, Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico, Milan, Italy; ⁸Department of Civil, Chemical, Environmental and Materials Engineering (DICAM), University of Bologna, Italy.

Self-assembled neuromuscular organoids (NMOs) contain functional neuronal and muscular compartments that can be derived from induced pluripotent stem cells (hiPSCs). Here we implemented a decellularized skeletal muscle (dSkM) scaffold to derive functional and physiologically active tissue-engineered NMOs (t-NMOs) from hiPSCs. Physiologically relevant activities of t-NMOs could be quantified using live imaging analysis. We also validated our platform using Duchenne muscular dystrophy (DMD) patient-derived hiPSCs to produce DMD t-NMOs. Upon neuronal stimulation, DMD t-NMOs mimic the reduced SkM contraction and altered calcium dynamics typical of the disease. Finally, we used the dSkM to study the role of native extracellular matrix (ECM) on hiPSC commitment towards neuromesodermal progenitors (NMPs), showing that dSkM was sufficient to modulate hiPSCs commitment. Altogether, our study described a patient-derived tissue-engineered organoid platform that model the human neuromuscular system (dys)functionality, and opens new perspectives for future understanding of the ECM role during NMO derivation.