149VP 3D stem cell-derived spinal cord/muscle organoid model for studying and treating neuromuscular diseases

• Published in: Neuromuscular disorders : NMD 2024-10, Vol.43, p.104441, Article 104441.584
• Main Authors: Corti, S., D' Angelo, A., Beatrice, F., Ongaro, J., Rinchetti, P., Faravelli, I., Miotto, M., Lodato, S., Nizzardo, M., Comi, G., Ottoboni, L.
• Format: Article
• Language: English
• ISSN: 0960-8966
• DOI: 10.1016/j.nmd.2024.07.593

This research aims to create a three-dimensional (3D) organoid model of the spinal cord and skeletal muscle from stem cells for the investigation and potential treatment of neuromuscular diseases, specifically spinal muscular atrophy (SMA) and amyotrophic lateral sclerosis (ALS). The focus is on understanding comparative and different pathological features and disease mechanisms, ultimately to evaluate therapeutic strategies. SMA and ALS are severe neurological disorders marked by the gradual deterioration of motor neurons. Establishing robust human models is crucial for enhancing our knowledge of and identifying therapeutic approaches for these neuromuscular disorders, which are known for causing paralysis and are attributed to disruptions in the complex network that controls voluntary movement. We created organoids representing the human spinal cord and muscle tissue using induced pluripotent stem cells (iPSCs) taken from patients with SMA, ALS C9ORF72 (C9) mutations, and non-affected individuals. Our methods of analysis included comprehensive transcriptomics, single-cell RNA sequencing, multielectrode array assessments, and immunophenotyping. As regards pharmacological intervention, SMA organoids were treated with a Risdiplam-like compound while C9 organoids with an antisense oligonucleotide with morpholino chemistry with proved efficacy in the murine counterpart of the C9 mouse model as C9-derived dipeptide modulator. So far, our organoid models for SMA and C9-ALS displayed significant differences in cellular and molecular development compared to controls, involving various cell types and extending beyond motor neurons, indicating a developmental aspect to both diseases. Specifically, for SMA organoids, treatment with the Risdiplam-like compound resulted an efficient correction of the proportion of full-length to truncated SMN2 protein accompanied by a modulation of about 15% of the genes, showcasing significant in vitro durability. Importantly, this treatment corrected and reversed multiple pathological indicators within the SMA organoids. The organoids proved effective for studying drug interactions and the impact of therapies, highlighting the profound and early developmental influence of SMA and C9ALS. Our results promote the potential of Risdiplam-like therapies in treating extensive facets of SMA pathology and emphasize the need of deeply understanding the pharmacodynamics involved. This research advances the prospect of refining therapeutic appr