Human three-dimensional engineered muscle tissue characterization and intramyocellular lipid modeling
Ontology highlight
ABSTRACT:
ORGANISM(S): Homo sapiens
PROVIDER: GSE345454 | GEO | 2026/08/31
REPOSITORIES: GEO
Similar Datasets
Project description:Aims/hypothesis: While lipid deposition in skeletal muscle is considered to be involved in obesity-associated insulin resistance, neutral intramyocellular lipid (IMCL) accumulation per se does not necessarily induce insulin resistance. We previously demonstrated that overexpression of the lipid droplet coat protein perilipin 2 augments intramyocellular lipid content while improving insulin sensitivity. Another member of the perilipin family, perilipin 5 (PLIN5), is predominantly expressed in oxidative tissues like skeletal muscle. Here we investigated the effects of PLIN5 overexpression – in comparison with effects of PLIN2 – on skeletal muscle lipid levels, gene expression profiles and insulin sensitivity. Methods: Gene electroporation was used to overexpress PLIN5 in tibialis anterior muscle of rats fed a high fat diet. Eight days after electroporation, insulin-mediated glucose uptake in skeletal muscle was measured by means of a hyperinsulinemic euglycemic clamp. Electron microscopy, fluorescence microscopy and lipid extractions were performed to investigate IMCL accumulation. Gene expression profiles were obtained using microarrays. Results: TAG storage and lipid droplet size increased upon PLIN5 overexpression. Despite the higher IMCL content, insulin sensitivity was not impaired and DAG and acylcarnitine levels were unaffected. In contrast to the effects of PLIN2 overexpression, microarray data analysis revealed a gene expression profile favoring FA oxidation and improved mitochondrial function. Conclusions/interpretation: Both PLIN2 and PLIN5 increase neutral IMCL content without impeding insulin-mediated glucose uptake. As opposed to the effects of PLIN2 overexpression, overexpression of PLIN5 in skeletal muscle promoted expression of a cluster of genes under control of PPARα and PGC1α involved in FA catabolism and mitochondrial oxidation.
2013-01-29 | GSE43832 | GEO
Project description:The accumulation of intramyocellular lipid (IMCL) is recognized as an important determinant of insulin resistance, and is increased by a high-fat diet (HFD). However, the effects of HFD on IMCL and insulin sensitivity are highly variable. We used microarrays to detail to identify the genes in skeletal muscle that are related to inter-individual variation of the effects with HFD on the accumulation of intramyocellular lipid and insulin sensitivity.
2015-05-01 | GSE68231 | GEO
Project description:Background Three-dimensional (3D) bioengineered models of human skeletal muscle are a promising approach for studying muscle development, function and disease in vitro. These models more closely resemble the complexity of native muscle tissue than two-dimensional (2D) monolayer culture and allow for functional measurements to be performed. However, a more complete understanding of how culture condition and duration impacts the myotube maturity and function is required to validate the transition from 2D to 3D culture of muscle cells. Methods Human skeletal muscle cells were cultured as either 2D monolayers or within 3D fibrin-based hydrogels as muscle constructs for up to 21 days. Quantitative proteomic analysis and functional assessments, including contractile force and cross-sectional area measurements, were conducted to evaluate the impact of culture conditions and duration on muscle cell differentiation. Results Proteomic analysis revealed myoblasts differentiated into myotubes by 8 days of differentiation in both 2D and 3D environments. However, the proteomic profiles of myotubes varied significantly between the two culture environments. At day 8 of differentiation, myosin heavy chain isotype abundance indicated a predominantly slow-twitch phenotype in 3D constructs, compared to a mixed fibre type phenotype in 2D monolayers. By day 21 of differentiation, 3D muscle constructs displayed improved mitochondrial maturity, extracellular matrix remodelling, and signs of transitioning towards a fast-twitch phenotype. This prolonged culture duration also resulted in increased passive tension but decreased peak contractile force in 3D muscle constructs. Conclusions This study demonstrates that 3D culture promotes maturity in human skeletal muscle cells, mimicking the biochemical cues and energy demands seen in native muscle tissue. The data highlights the importance of selecting appropriate culture conditions and durations when studying skeletal muscle cells in vitro and suggests 8 days of differentiation as optimal for achieving peak contractile force in 3D muscle constructs.
2025-06-23 | PXD061227 | Pride
Project description:Aims/hypothesis: While lipid deposition in skeletal muscle is considered to be involved in obesity-associated insulin resistance, neutral intramyocellular lipid (IMCL) accumulation per se does not necessarily induce insulin resistance. We previously demonstrated that overexpression of the lipid droplet coat protein perilipin 2 augments intramyocellular lipid content while improving insulin sensitivity. Another member of the perilipin family, perilipin 5 (PLIN5), is predominantly expressed in oxidative tissues like skeletal muscle. Here we investigated the effects of PLIN5 overexpression M-bM-^@M-^S in comparison with effects of PLIN2 M-bM-^@M-^S on skeletal muscle lipid levels, gene expression profiles and insulin sensitivity. Methods: Gene electroporation was used to overexpress PLIN5 in tibialis anterior muscle of rats fed a high fat diet. Eight days after electroporation, insulin-mediated glucose uptake in skeletal muscle was measured by means of a hyperinsulinemic euglycemic clamp. Electron microscopy, fluorescence microscopy and lipid extractions were performed to investigate IMCL accumulation. Gene expression profiles were obtained using microarrays. Results: TAG storage and lipid droplet size increased upon PLIN5 overexpression. Despite the higher IMCL content, insulin sensitivity was not impaired and DAG and acylcarnitine levels were unaffected. In contrast to the effects of PLIN2 overexpression, microarray data analysis revealed a gene expression profile favoring FA oxidation and improved mitochondrial function. Conclusions/interpretation: Both PLIN2 and PLIN5 increase neutral IMCL content without impeding insulin-mediated glucose uptake. As opposed to the effects of PLIN2 overexpression, overexpression of PLIN5 in skeletal muscle promoted expression of a cluster of genes under control of PPARM-NM-1 and PGC1M-NM-1 involved in FA catabolism and mitochondrial oxidation. Rats received a high fat diet for 3 weeks; 2 weeks after start of the diet intervention Plin5 (OXPAT) or Plin2 (ADRP) were overexpressed in either the right or left tibialis anterior muscle. One week later pooled tibialis anterior muscle samples were analysed on microarrays.
2013-01-29 | E-GEOD-43832 | biostudies-arrayexpress
Project description:The ability to recapitulate muscle differentiation in vitro has proven invaluable to investigate mechanisms of myogenesis, muscle cell function and muscle diseases. However, obtaining myoblasts from patients with neuromuscular diseases poses ethical and procedural challenges which limit investigations of molecular mechanisms of muscle pathophysiology. Alternative myogenic models have been developed, such as the derivation of myogenic cells from skin fibroblasts through activation of an endogenous myogenic program triggered by exogenous expression of murine Myod. In the context of this RNA-seq dataset, we compare the transcriptome of myo-converted human fibroblasts and isogenic in vitro differentiated myoblasts. We show that myogenic induction of fibroblasts elicits genome-wide transcriptomic changes indicative of strong myogenic commitment and differentiation, including marked upregulation of genes implicated in cell cycle exit and in several myogenic pathways. Yet, we find that myotubes are further along myogenic commitment than myo-converted fibroblasts under the conditions tested. Extension of myo-conversion to 7 days does not significantly enhance myogenic commitment, from a gene expression standpoint. This suggests that myo-converted fibroblasts and myotubes retain some cell type specificity of gene expression profiles. Although these myogenic cell types are not identical, our results nonetheless favor a view of myo-converted fibroblasts as a robust and practical model to investigate cellular and genomic properties of cells from patients with muscle pathologies.
2023-08-21 | GSE236118 | GEO
Project description:The ability to recapitulate muscle differentiation in vitro has proven invaluable to investigate mechanisms of myogenesis, muscle cell function and muscle diseases. However, obtaining myoblasts from patients with neuromuscular diseases poses ethical and procedural challenges which limit investigations of molecular mechanisms of muscle pathophysiology. Alternative myogenic models have been developed, such as the derivation of myogenic cells from skin fibroblasts through activation of an endogenous myogenic program triggered by exogenous expression of murine Myod. In the context of this ChIP-seq dataset, we compared the transcriptome and lamina-associated domains (LADs) as genome organizers in myo-converted human fibroblasts and in isogenic myotubes differentiated from myoblasts. We show that myogenic induction of fibroblasts elicits genome-wide transcriptomic changes indicative of myogenic commitment and differentiation. Yet, myotubes are further along myogenic commitment than myo-converted fibroblasts under the conditions tested. LADs with typical LAD properties (low gene density, containing mostly repressed genes, and H3K9me3-rich) are shared between the two cell types, but each cell type also displays nearly 700 unique LADs. Strikingly, myotube-specific LADs are smaller, more gene-rich and less heterochromatic than shared LADs or LADs unique to myo-converted fibroblasts. Thus, myo-converted fibroblasts and myotubes retain some cell type specificity of genome organization at this level. Although these myogenic cell types are not identical, our results favor a view of myo-converted fibroblasts as a robust and practical model to investigate cellular and genomic properties of cells from patients with muscle pathologies.
2023-08-21 | GSE236115 | GEO
Project description:The accumulation of intramyocellular lipid (IMCL) is recognized as an important determinant of insulin resistance, and is increased by a high-fat diet (HFD). However, the effects of HFD on IMCL and insulin sensitivity are highly variable. We used microarrays to detail to identify the genes in skeletal muscle that are related to inter-individual variation of the effects with HFD on the accumulation of intramyocellular lipid and insulin sensitivity. In order to identify the gene expression profile related with inter-individual variation of the effects with high fat diet on the accumulation of intramyocellular lipid and insulin sensitivity, we classified subjects who showed a large increase in intramyocellular lipid and a large decrease in glucose infusion rate by high fat diet as the high-responder (HR), and the subjects who showed a small increase in intramyocellular lipid and a small decrease in glucose infusion rate were classified as the low-responder (LR). In 5 subjects in each group, skeletal muscle sample selected at before (pre) and after (post) 3days high fat diet for RNA extraction and hybridization on Affymetrix microarrays.
2015-05-01 | E-GEOD-68231 | biostudies-arrayexpress
Project description:The study was designed to compare whether and how muscle fibroblasts (FIB) and muscle stem cells (MYO) interact with motor neurons. Moreover, we aimed to investigate whether muscle cells, isolated from lifelong exercisers, exerted a protective and supportive effect on motor neurons. These objectives were investigated in vitro using primary muscle cells from human donors and primary motor neurons from rat embryos and were analyzed by immunocytochemistry and species-specific RNA sequencing. Rat neuron cells were treated for 24 hours with conditioned media from human skeletal muscle fibroblasts and myoblasts.
2026-02-21 | E-MTAB-14399 | biostudies-arrayexpress
Project description:Muscle is highly hierarchically organized, with functions shaped by genetically controlled expression of protein ensembles with different isoform profiles at the sarcomere scale. However, it remains unclear how isoform profiles shape whole-muscle performance. We compared two mouse hindlimb muscles, the slow, relatively parallel-fibered soleus and the faster, more pennate-fibered tibialis anterior (TA), across scales: from gene regulation, isoform expression and translation speed, to force-length-velocity-power for intact muscles. Expression of myosin heavy-chain (MHC) isoforms directly corresponded with contraction velocity. The fast-twitch TA with fast MHC isoforms had faster unloaded velocities (actin sliding velocity, Vactin; peak fiber velocity, Vmax) than the slow-twitch soleus. For the soleus, Vactin was biased towards Vactin for purely slow MHC I, despite this muscle's even fast and slow MHC isoform composition. Our multi-scale results clearly identified a consistent and significant dampening in fiber shortening velocities for both muscles, underscoring an indirect correlation between Vactin and fiber Vmax that may be influenced by differences in fiber architecture, along with internal loading due to both passive and active effects. These influences correlate with the increased peak force and power in the slightly more pennate TA, leading to a broader length range of near-optimal force production. Conversely, a greater force-velocity curvature in the near-parallel fibered soleus highlights the fine-tuning by molecular-scale influences including myosin heavy and light chain expression along with whole-muscle characteristics. Our results demonstrate that the individual gene, protein and whole-fiber characteristics do not directly reflect overall muscle performance but that intricate fine-tuning across scales shapes specialized muscle function.
2022-05-23 | GSE203400 | GEO
Project description:Skeletal muscle aging is associated with increased lipid accumulation, or myosteatosis, leading to lipotoxicity and loss of muscle function. Here, we report that loss of the novel lactate receptor GPR81 in aged muscle is associated with impaired lipid oxidation and enhanced lipid accumulation. Knockdown of GPR81 in young healthy myoblasts led to an increase in senescence hallmarks such as DNA damage, accumulation of reactive oxygen species (ROS), mitochondrial dysfunction and impaired autophagy. Conversely, treatment of senescent myoblasts with GPR81 agonists enhanced lipid oxidation in senescent myoblasts leading to a decrease in lipid accumulation, ultimately resulting in decreased DNA damage, ROS accumulation and enhanced ability to form myotubes. In agreement with our in vitro findings, we observed significant improvement in muscle regeneration and overall health of aged or progeric mice that were treated with GPR81 agonists. Our findings suggest that GPR81 plays a key role in skeletal muscle lipid metabolism, and agonists of GPR81 might play a promising role in reversing age-associated lipid accumulation and loss of muscle function.
2026-06-01 | GSE325224 | GEO