Native-extracellular matrix drives human neuromuscular organoid morphogenesis and functionality
Ontology highlight
ABSTRACT:
PROVIDER: PRJNA940416 | ENA |
REPOSITORIES: ENA
Similar Datasets
Project description:Human neuromuscular organoids (NMOs) hold a great potential to study the pathophysiology of functional human skeletal muscle in vitro due to the presence of both muscular and neuronal compartments organized in a their three-dimensional (3D) context and to be derived from induced pluripotent stem cells (hiPSCs). An open question in the field remains how the morphology of NMOs can influence the maturation and functionality of the neuronal-coupled skeletal muscle. NMOs self-assemble into spheroids, thus not fully recapitulating the morphological organization of the innervated skeletal muscle. Proper development, maturation and function of the innervated skeletal muscle require defined 3D organization of the cells in a context of tissue-specific extracellular matrix (ECM). We hypothesized that extracellular structural imprinting along hiPSC differentiation could provide self-assembly guidance cues able to impact on NMO morphogenesis and on neuronal-coupled skeletal muscle maturation and functionality. Therefore, here we used a well characterized skeletal muscle ECM to guide the morphogenesis of differentiating hiPSC toward tissue-like structured NMOs (t-NMOs). T-NMOs resemble the morphological organization of the neuronal-coupled skeletal muscle, with elongated bundles of myofibers reached by long neural projections (up to 1mm length). In presence of the instructive native ECM, high degree of skeletal muscle maturation together with functional neuromuscular junctions were reached in t-NMOs in only 30 days. Finally, we used t-NMO to understand whether neuronal activation could lead to altered myogenic response in a pathologic context. To do so, we used hiPSCs derived from patients affected by Duchenne Muscular Dystrophy (DMD), a neuromuscular disease characterized by myofiber weakness and membrane fragility, as well as altered calcium handling. Upon neuronal stimulation, DMD t-NMO displayed key muscular phenotypes of the disease, such as altered functionality and calcium dynamics, as well as myofiber rupture.
2025-05-30 | GSE226477 | GEO
Project description:A cultured sensorimotor organoid model forms human neuromuscular junctions
| PRJNA732479 | ENA
Project description:A cultured sensorimotor organoid model forms human neuromuscular junctions
| PRJNA732480 | ENA
Project description:Vascularization of cardiac organoids controls the extracellular matrix environment and regulates organoid functionality
| PRJNA634359 | ENA
Project description:Homeostatic mini-intestines through scaffold-guided organoid morphogenesis
| PRJNA624065 | ENA
Project description:Crosstalk between cardiac cells is critical during heart development but its role in organ maturation is still largely uncharacterised. Here, we show that endothelial cells increase the force of contraction and enhance the expression of mature sarcomeric proteins and extracellular matrix (ECM) components in human pluripotent stem cell derived cardiac organoids (hCO). Endothelial cells regulate cardiac maturation and function both directly through secretion of ECM molecules and indirectly via paracrine signaling. Laminin α5, an endothelial enriched ECM protein, was identified as a key regulator of cardiac maturation and contractility in vitro. In vivo loss-of-function studies in mice confirmed that Lama5 was required for myocardial expansion during heart development in vivo. In addition, paracrine PDGF signaling was identified as a mediator of increased ECM deposition and cardiac contractility in hCO. This study uncovers matrix regulatory functions of endothelial cells governing cardiac maturation and highlights the importance of multicellularity for organoid models.
2023-05-20 | GSE150998 | GEO
Project description:Muscle fibroadipogenic progenitor (FAP) cells, which are muscular mesenchymal cells that originate from lateral plate mesoderm have been proposed to act as a critical regulator for adult muscle homeostasis1–7, including the maturation and proper functioning of the neuromuscular junction (NMJ)3, Prx-Bap1 paper. However, the mechanism and intercellular crosstalk by which FAPs regulate the stability and functionality of neuromuscular system remains unknown. Here we show that FAPs not only locally but also systemically regulate the neuromuscular system through the secretion of a serine-type endopeptidase Granzyme E which may imply the previously unidentified endocrine function of FAPs. Local transplantation of wild-type FAPs into the neuromuscular disease model (Prrx1Cre;Bap1f/f, hereafter, cKO) can readily prevent neuromuscular defects, including degeneration of the neuromuscular junction and loss of motor neurons. These effects are found not only in transplanted hindlimb muscles but also in the contralateral hindlimb and even forelimb muscles. Notably, subcutaneous administration of microparticles encapsulating FAP-conditioned media into cKO mice was sufficient to restore normal neuromuscular functions. By analyzing the transcriptomic and secretomic profiles of FAPs, we identified a novel protein, Granzyme E, which is specifically expressed in and secreted by FAPs, and which indispensably regulates the structure and function of NMJ and motor neuron survival. Our study has defined a unique mechanism of Granzyme E-dependent, systemic control of the neuromuscular system by FAPs, which would provide a comprehensive understanding on the neuromuscular systems and their crosstalk with non-neuronal cells. These findings may provide a therapeutic benefit to treat NMJ-related diseases.
2022-08-31 | GSE199660 | GEO
Project description:Leigh syndrome (LS) is a severe manifestation of mitochondrial disease in children and is currently incurable. The lack of effective models hampers our understanding of the mechanisms underlying the neuronal pathology of LS. Using patient-derived induced pluripotent stem cells and CRISPR/Cas9 engineering, we developed a human model of LS due to mutations in the complex IV assembly gene SURF1. Single-cell RNA-sequencing and multi-omics analysis revealed compromised neuronal morphogenesis in mutant neural cultures and brain organoids. The defects emerged at the level of neural progenitor cells (NPCs), which retained a glycolytic proliferative state that failed to instruct neuronal morphogenesis. LS NPCs carrying mutations in the complex I gene NDUFS4 recapitulated morphogenesis defects. Interventions supporting the metabolic programming of NPCs restored neuronal morphogenesis, including SURF1 gene augmentation and PGC1A induction via bezafibrate treatment. Our findings provide mechanistic insights and suggest interventional strategies for a rare mitochondrial disease with major unmet medical needs.
2021-01-29 | PXD019112 | Pride
Project description:Neuromuscular networks assemble during early human embryonic development and are essential for the control of body movement. Previous neuromuscular junction modeling efforts using human pluripotent stem cells (hPSCs) generated either spinal cord neurons or skeletal muscles in monolayer culture. Here, we use hPSC-derived axial stem cells, the building blocks of the posterior body, to simultaneously generate spinal cord neurons and skeletal muscle cells that self-organize to generate human neuromuscular organoids (NMOs) that can be maintained in 3D for several months. Single-cell RNA-sequencing of individual organoids revealed reproducibility across experiments and enabled the tracking of the neural and mesodermal differentiation trajectories as organoids developed and matured. NMOs contain functional neuromuscular junctions supported by terminal Schwann cells. They contract and develop central pattern generator-like neuronal circuits. Finally, we successfully use NMOs to recapitulate key aspects of myasthenia gravis pathology, thus highlighting the significant potential of NMOs for modeling neuromuscular diseases in the future.
2020-01-20 | GSE128357 | GEO
Project description:Muscle denervation due to injury, disease or aging results in impaired motor function. Restoring neuromuscular communication requires axonal regrowth and regeneration of neuromuscular synapses. Muscle activity inhibits neuromuscular synapse regeneration. The mechanism by which muscle activity regulates regeneration of synapses is poorly understood. Dach2 and Hdac9 are activity-regulated transcriptional co-repressors that are highly expressed in innervated muscle and suppressed following muscle denervation. Here, we report that Dach2 and Hdac9 inhibit regeneration of neuromuscular synapses. Importantly, we identified Myog and Gdf5 as muscle-specific Dach2/Hdac9-regulated genes that stimulate neuromuscular regeneration in denervated muscle. Interestingly, Gdf5 also stimulates presynaptic differentiation and inhibits branching of regenerating neurons. Finally, we found that Dach2 and Hdac9 suppress miR206 expression, a microRNA involved in enhancing neuromuscular regeneration. RNAseq on innervated and 3 day denervated adult soleus muscle from wildtype mice is compared with that from 3 day denervated soleus muscle from Dach2/Hdac9 deleted mice to identify Dach2/Hdac9-regulated genes.
2015-10-29 | E-GEOD-58669 | biostudies-arrayexpress