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Single-cell RNA sequencing reveals maturation trajectory in human pluripotent stem cell-derived cardiomyocytes in engineered tissues

ABSTRACT: Single-cell RNA sequencing reveals maturation trajectory in human pluripotent stem cell-derived cardiomyocytes in engineered tissues

PROVIDER: PRJNA823929 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR18653949.fastq.gz	Fastqsanger.gz
	SRR18653950.fastq.gz	Fastqsanger.gz
	SRR18653951.fastq.gz	Fastqsanger.gz
	SRR18653952.fastq.gz	Fastqsanger.gz
	SRR18653953.fastq.gz	Fastqsanger.gz

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Single-cell RNA sequencing reveals maturation trajectory in human pluripotent stem cell-derived cardiomyocytes in engineered tissues

Project description:Cardiac in vitro models have become increasingly easy and affordable with the optimisation of stem cell-based cardiomyocyte (CM) differentiation. However, the CMs obtained are still far from being as mature as their in vivo counterparts. Here we study the cellular phenotype of human pluripotent stem cell-derived CMs by comparing the single-cell level gene expression profiles among different engineered cardiac tissue configurations: human ventricular cardiac anisotropic sheet, human ventricular cardiac tissue strip, and human ventricular cardiac organoid chamber (hvCOC). As a control, the spontaneously formed 3D cardiac spheroid aggregates (CS) were used. Our results suggest that the level of maturation increases with the level of complexity of the engineered tissues with CS being the least mature and hvCOC the most mature tissue. We show that the contractile components are the first function to mature, followed by electrophysiology and oxidative metabolism. Notably, the cellular structures in the 2D constructs show higher organisation while metabolic maturity preferentially increase in the 3D structures. We conclude that more complex tissue engineering models could offer more physiological and reliable in vitro matured cardiac tissue model for drug screening or disease modelling.

2023-03-08 | GSE200277 | GEO

Metabolomics Identifies Metabolic Markers of Maturation in Human Pluripotent Stem Cell-Derived Cardiomyocytes

Project description:Cardiovascular disease is a leading cause of death worldwide. Human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) hold immense clinical potential and recent studies have enabled generation of virtually pure hPSC-CMs with high efficiency in chemically defined and xeno-free conditions. Despite these advances, hPSC-CMs exhibit an immature phenotype and are arrhythmogenic in vivo, necessitating development of methods to mature these cells. hPSC-CMs undergo significant metabolic alterations during differentiation and maturation. A detailed analysis of the metabolic changes accompanying maturation of hPSC-CMs may prove useful in identifying new strategies to expedite the maturation process and also provide biomarkers for testing or validating hPSC-CM maturation. In this study we identified global metabolic changes which take place during long-term culture and maturation of hPSC-CMs derived from three different hPSC lines. We have identified several metabolic pathways, including phospholipid metabolism and pantothenate and Coenzyme A metabolism, which showed significant enrichment upon maturation in addition to fatty acid oxidation and metabolism. We also identified an increase in glycerophosphocholine and reduction in phosphocholine as potential metabolic biomarkers of maturation. These biomarkers were also affected in a similar manner during murine heart development in vivo. These results support that hPSC-CM maturation is associated with extensive metabolic rewiring and understanding the role of these metabolic changes in maturation process has the potential to develop novel approaches to monitor and expedite hPSC-CM maturation.

2018-04-24 | MTBLS425 | MetaboLights

Single-cell RNA sequencing reveals maturation trajectory in human pluripotent stem cell-derived cardiomyocytes in engineered tissues.

Project description:Cardiac in vitro models have become increasingly obtainable and affordable with the optimization of human pluripotent stem cell-derived cardiomyocyte (hPSC-CM) differentiation. However, these CMs are immature compared to their in vivo counterparts. Here we study the cellular phenotype of hPSC-CMs by comparing their single-cell gene expression and functional profiles in three engineered cardiac tissue configurations: human ventricular (hv) cardiac anisotropic sheet, cardiac tissue strip, and cardiac organoid chamber (hvCOC), with spontaneously aggregated 3D cardiac spheroids (CS) as control. The CM maturity was found to increase with increasing levels of complexity of the engineered tissues from CS to hvCOC. The contractile components are the first function to mature, followed by electrophysiology and oxidative metabolism. Notably, the 2D tissue constructs show a higher cellular organization whereas metabolic maturity preferentially increases in the 3D constructs. We conclude that the tissue engineering models resembling configurations of native tissues may be reliable for drug screening or disease modeling.

| S-EPMC10025988 | biostudies-literature

Unbiased Proteomics Benchmarking of Maturation of Human Pluripotent Stem Cell-Derived Cardiomyocytes

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

Mef2c/p300 Promotes Maturation of Induced Cardiomyocytes

Project description:Mef2c/p300 Promotes Maturation of Induced Cardiomyocytes

| PRJNA923690 | ENA

Contractile work contributes to maturation of energy metabolism in hiPSC-derived cardiomyocytes

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

PPARdelta signaling induces metabolic maturation in human pluripotent stem cell-derived cardiomyocytes

Project description:PPARdelta signaling induces metabolic maturation in human pluripotent stem cell-derived cardiomyocytes

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Engineered vasculature induces functional maturation of pluripotent stem cell-derived islet organoids

Project description:Engineered vasculature induces functional maturation of pluripotent stem cell-derived islet organoids

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Gene expression analysis of human induced pluripotent stem cell-derived (hiPSC) cardiomyocytes in 2D versus 3D (Engineered heart tissue) format

Project description:Analysis of cardiomyocytes cultivated in 2D or age-matched 3D (Engineered heart tissue, EHT) format.

2016-04-20 | GSE80390 | GEO

Lactate and Immunomagnetic-purified iPSC-derived Cardiomyocytes Generate Comparable Engineered Cardiac Tissue Constructs

Project description:top down and bottom up proteomics files for Lactate and Immunomagnetic-purified iPSC-derived Cardiomyocytes Generate Comparable Engineered Cardiac Tissue Constructs

2023-05-05 | MSV000091869 | MassIVE

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