Perlecan (HSPG2) signalling promotes structural, contractile, and metabolic development during the maturation stage of hPSC-CM differentiation
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ABSTRACT:
PROVIDER: PRJNA956913 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR24204143_1.fastq.gz | Fastqsanger.gz | |||
| SRR24204143_2.fastq.gz | Fastqsanger.gz | |||
| SRR24204144_1.fastq.gz | Fastqsanger.gz | |||
| SRR24204144_2.fastq.gz | Fastqsanger.gz | |||
| SRR24204145_1.fastq.gz | Fastqsanger.gz |
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Project description:Perlecan (HSPG2), a multifunctional heparan sulphate proteoglycan (HSPG), is a key player in extracellular matrix maturation and stabilisation. Structurally perlecan is similar to agrin, another HSPG known to contribute to cardiomyocyte phenotype switching by reverting hypertrophy. Although perlecan is crucial for cardiac development, its role in cardiomyocyte phenotypic switching is unknown. Here we show perlecan expression increases over time during the differentiation of pluripotent stem cells to cardiomyocytes (hPSC-CMs). Haploinusuffient hPSCs (HSPG2+/-) differentiate efficiently towards cardiomyocytes with minimal transcriptomic differences seen early in differentiation, but wide-ranging changes seen in late-stage cardiomyocytes. These differences are associated with structural, contractile, metabolic, and ECM genes. In keeping, late-stageHSPG2+/-hPSC-CMs display reduced maximal metabolic capacity and increased extracellular acidification rate, characteristics of reduced maturity. Moreover, engineered heart tissues produced withHSPG2+/-hPSC-CMs exhibit increased ECM remodelling, reduced tissue thickness and force generation. Wild-type hPSC-CMs grown on a substrate coated with a perlecan peptide showed increased cardiomyocyte nucleation typical of hypertrophic growth, suggesting that perlecan plays the opposite role of agrin by promoting cellular maturation rather than hyperplasia and proliferation. These data show that perlecan promotes cardiomyocyte maturity, possibly through hypertrophic growth. Targeting perlecan-dependent signalling may, therefore, reverse the phenotypic switch common to many forms of heart failure, by improving cardiomyocyte functionality.
2024-01-23 | GSE229975 | GEO
Project description:Energy metabolism is a key aspect of cardiomyocyte biology. Human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CM) are a promising tool for biomedical application, but they are immature and have not undergone metabolic shift related to early postnatal development. Cultivation of hiPSC-CM in 3D engineered heart tissue (EHT) format leads to morphological maturation. This study compared the mitochondrial and metabolic state of hiPSC-CM in standard 2D culture and the EHT format and determined the influence of contractile activity. HiPSC-CM in EHTs showed ~2-fold higher number of mitochondria (electron microscopy), mitochondrial mass (mitotracker), DNA (Mt-ND1, Mt-ND2), and protein abundance (proteome) than in 2D culture. While hiPSC-CM exhibited the principal ability to use glucose, lactate and fatty acids as energy substrates irrespective of culture format, hiPSC-CM in 3D performed more oxidation of glucose, lactate and fatty acid, and less anaerobic glycolysis. The increase in mitochondrial mass and DNA in 3D was diminished by pharmacological inhibition of contractile force, suggesting that contractile work participates in mitochondrial development hiPSC-CM. In conclusion, contractile work in the EHT format contributes to metabolic maturation of hiPSC-CM.
2018-03-06 | PXD008205 | Pride
Project description:Perlecan is a critical component of basement membrane proteins in tissue development. However, there are few studies investigating direct impact of perlecan on mesenchymal tissue differentiation using genetic modification (gain- and loss-of-function mutations) and epigenetic modification (matrix microenvironment). In this report, Cas9-mediated knockout (KO) of heparin sulfate proteoglycan 2 (HSPG2) and overexpression (OE) in human fetal nucleus pulposus stem/progenitor cells (NPSCs) and adult infrapatellar fat pad-derived stem cells (IPFSCs) were evaluated for the influence on mesenchymal differentiation. Furthermore, decellularized ECM (dECM) deposited by fetal NPSCs with either HSPG2 KO or OE was evaluated for its rejuvenation effect on adult NPSCs in proliferation and mesenchymal differentiation. We found that adult IPFSCs confirmed the unique role of perlecan in mesenchymal differentiation of fetal NPSCs; dECM with perlecan overexpression significantly promoted adult NPSCs in chondrogenic capacity rather than osteogenic and adipogenic capacities, indicating a potential application of perlecan in engineering smart biomaterial for cartilage regeneration.
2025-03-26 | GSE275764 | GEO
Project description:Rationale: Human pluripotent stem cells-derived cardiomyocytes (hPSC-CMs) exhibit the properties of fetal CMs, which limit their applications. Various methods have been used to promote maturation of hPSC-CMs; however, there is a lack of an unbiased and comprehensive method for accurate benchmarking of hPSC-CM maturation.
Objective: We aim to develop an unbiased proteomics method integrating high-throughput top-down targeted proteomics and bottom-up global proteomics for accurate and comprehensive assessment of hPSC-CM maturation.
Methods and Results: Utilizing hPSC-CMs from early- and late-stage two-dimensional monolayer culture and three-dimensional engineered cardiac tissue, we demonstrated high reproducibility and reliability of the top-down proteomics method, which enabled simultaneous quantification of contractile protein isoform expressions and their PTMs. This method allowed for the detection of known maturation-associated contractile protein alterations, and for the first time, identified contractile protein PTMs as promising new markers of maturation. By employing a global proteomics strategy, we identified candidate maturation markers important for sarcomere organization, cardiac excitability, and Ca2+ homeostasis; and validated these markers in the developing mouse cardiac ventricles.
Conclusions: We established an unbiased proteomics method that can provide accurate and specific benchmarking of hPSC-CM maturation, and identified new markers of maturation. Furthermore, this integrated proteomics strategy laid a strong foundation for uncovering molecular basis underlying cardiac development and disease using hPSC-CMs.
| MSV000082985 | MassIVE
Project description:Chloromethanomonas sp. CM strain:CM Genome sequencing
| PRJNA678572 | ENA
Project description:Stem cells reside in specialised microenvironments or niches that balance stem cell proliferation and differentiation. The extracellular matrix (ECM) is an essential component of most niches, as it controls niche homeostasis, provides physical support and conveys extracellular signals. Basement membranes (BMs) are thin ECM sheets constituted mainly by Laminins, Perlecan, Collagen IV and Entactin/Nidogen and that surround epithelia and other tissues. Perlecans are secreted proteoglycans that interact with ECM proteins, ligands, receptors and growth factors such as FGF, PDGF, VEGF, Hedgehog and Wingless. Thus, Perlecans have structural and signalling functions through the binding, storage or sequestering of specific ligands. We have used the Drosophila ovary to assess the importance of Perlecan in the functioning of a stem cell niche. Ovarioles in the adult ovary are enveloped by an ECM sheath and possess a tapered structure at their anterior apex termed the germarium. The anterior tip of the germarium hosts the germline niche, where two to four germline stem cells (GSCs) reside together with few somatic cells: terminal filament (TF) cells, cap cells (CpCs) and escort cells (ECs). We report that niche architecture in the developing gonad requires trol, that niche cells secrete an isoform-specific, Perlecan-rich interstitial matrix and that DE-cadherin-dependent stem cell-niche adhesion necessitates trol. Hence, we provide evidence to support a structural role for Perlecan in germline niche establishment during larval stages and in the maintenance of a normal pool of stem cells in the adult niche.
2021-03-31 | GSE164866 | GEO
Project description:Pluripotent stem cell-derived cardiomyocytes (PSC-CMs) provide an unprecedented opportunity to study human heart development and disease. A major caveat however is that they remain functionally and structurally immature in culture, limiting their potential for disease modeling and regenerative approaches. Here we address the question of how different metabolic pathways can be modulated in order to induce efficient cardiac maturation of hPSC-CMs. We show that PPAR signaling acts in an isoform-specific manner to balance the glycolysis and fatty acid oxidation (FAO) pathways. PPARd activation or inhibition results in efficient respective up- or down-regulation of the gene regulatory networks underlying FAO in hPSC-CMs. PPARd induction further increases mitochondrial and peroxisome content, enhances mitochondrial cristae formation and augments FAO flux. Lastly PPARd activation results in enhanced myofibril organization and increased numbers of bi-nucleated hPSC-CMs. Transient lactate exposure, commonly used in hPSC-CM purification protocols, induces an independent program of cardiac maturation, but when combined with PPARd activation equally results in a metabolic switch to FAO. In summary, we identify multiple axes of metabolic modifications of hPSC-CMs including a role for PPARdelta signaling in inducing the metabolic switch to FAO in hPSC-CMs. Our findings provide new opportunities to generate and use metabolically mature hPSC-CMs including for disease modeling and regenerative therapy.
2022-04-09 | GSE178984 | GEO
Project description:Haloplasma contractile SSD-17B genome sequencing
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Project description:Phaffomycetaceae sp. UFMG-CM-Y7052 isolate:UFMG-CM-Y7052 Genome sequencing
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