Screening in Human Cardiac Organoids Identifies a Requirement for the Mevalonate Pathway in Cardiomyocyte Proliferation
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PROVIDER: PRJNA438501 | ENA |
REPOSITORIES: ENA
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Project description:Purpose: Induction of endogenous proliferation is a promising strategy for cardiac regeneration. We find that GSK3 inhibition activates a cell cycle network whereas MST1 inhibition drives the mevalonate pathway, with synergistic activation of proliferation. However, all GSK3 inhibitors tested also reduce contractile force in hCO. We screened for small molecule activators of cardiomyocyte proliferation that did not alter contractile force in hCOs. This was overcome by screening of a boutique compound library, identifying a p38 inhibitor, which activated a cell cycle network without reducing force. The screen also identified a TGFBR inhibitor that induces the mevalonate pathway and can also synergise to activate proliferation. RNA sequencing was performed to investigate underlying mechanisms of action. Methods: RNA samples were processed with Illumina TruSeq Stranded mRNA Library prep kit selecting for poly(A) taled RNA following the manufacturer’s recommendations. Libraries were quantified with Qubit HS (ThermoFisher) and Fragment Analyzer (Advances Analytical Technologies) adjusted to the appropriate concentration for sequencing. Indexed libraries were pooled and sequenced at a final concentration of 1.8 pM on an Illumina NextSeq 500 high-output run using paired-end chemistry with 75 bp read length. The sequencing data was demultiplexed using Illumina bcl2fastq2-v2.17. The quality of the reads was assessed thanks to FastQC. The reads were then processed and mapped to the human genome hg38 using the Bcbio-nextgen framework version 1.0.3. The aligner used was HISAT2 2.0.5. Raw counts were normalised and analysed with DESeq2. Results: Analysis of RNAseq data lead to the understanding that compound 3 regulated the cell cycle network, which was similar to GSK3 inhibitors, whereas compound 65 regulated the mevalonate network, which was similarly to MST1 inhibitors. Conclusions: The current study adds to a growing body of evidence that alterations in cardiomyocyte metabolism may be a cause rather than a consequence of cardiomyocyte cell cycle arrest. Controlling cellular metabolism is emerging as a key strategy in the fight against cancer but the same pathways may conversely be required for the development of cardiac regenerative therapies.
2019-03-20 | GSE111853 | GEO
Project description:Re-induction of endogenous proliferation is a promising regeneration strategy for post-mitotic organs. For heart regeneration, we recently discovered that dual inhibition of both GSK3 and MST1 is required for proliferation in mature human cardiac organoids1. However, the mechanisms of action are not yet understood. Here, we de-convolute the mitogenic response using proteomics and report that GSK3 inhibition activates a cell cycle network whereas MST1 inhibition drives the mevalonate pathway. Screening of an additional 105 compounds identified a p38 inhibitor, which activated a cell cycle network, as well as an inhibitor of TGFBR, which activated the mevalonate pathway; combinatorial treatment with these two inhibitors also resulted in synergistic cell cycle activation. The mevalonate pathway which was consistently activated under the most robust proliferative conditions, is down-regulated during in vivo maturation when cardiomyocyte regenerative capacity ceases and inhibition of the mevalonate pathway abolished the myocyte proliferative response to mitogens. Taken together, our findings suggest that the mevalonate pathway synergises with mitogenic agents to enable mature cardiomyocytes to re-enter the cell cycle. These findings have important ramifications for development of cardiac regenerative therapeutics.
2019-03-12 | PXD009133 | Pride
Project description:The mammalian heart undergoes maturation during postnatal life to meet the increased functional requirements of the adult. However, the key drivers of this process remain poorly defined. We developed as 96-well screening platform, using human pluripotent stem cell derived cardiac organoids, to determine the molecular requirements for in vitro cardiomyocyte maturation. Here, we describe gene expression changes resulting from culturing human cardiac organoids in standard cell culture conditions and under optimized maturation conditions. We assessed our maturation conditions by comparing transcriptional changes of human cardiac organoids to RNA isolated from human heart. Interesting, analysis of these data revealed that a switch to fatty acid oxidative metabolism is a key governor of cardiomyocyte maturation and mature cardiac organoids were refractory to mitogenic stimuli.
2017-08-30 | GSE93841 | GEO
Project description: Not available
| S-EPMC5635889 | biostudies-literature
Project description:Since the proliferative capacity of cardiomyocytes is extremely limited in the adult mammalian hearts, the irreversible loss of cardiomyocytes following cardiac injury markedly reduces cardiac function, leading to cardiac remodeling and heart failure. However, the early neonatal mice have a strong ability in cardiomyocyte proliferation and cardiac regeneration after heart damage such as apical resection. Besides of cardiomyocytes, non-myocytes in heart tissue also play important roles in the regeneration process. Previous studies showed that cardiac macrophages, regulatory T cells and CD4+ T cells are all involved in regulating the myocardial regeneration process. However, the roles of other cardiac immune cells in cardiac regeneration remains to be elucidated. B cells is a prominent immune cell in injured heart; here we discovered the indispensable function of cardiac B cells in improving cardiomyocyte proliferation and heart regeneration in neonatal mice.
2023-06-02 | GSE205115 | GEO
Project description:In this study, we have investigated the effect of p53 deletion on the metabolic activity of colon cancer cells exposed to metabolic stress. In order to recreate the simultaneous reduction in oxygen and nutrient availability found in tumors, we cultured cancer cells as multicellular tumor spheroids. Under these conditions, p53 deficient cancer cells activate the expression of enzymes of the mevalonate pathway via the sterol regulatory element binding protein 2 (SREBP2). Moreover, inhibition of mevalonate pathway activity with statins selectively induced apoptosis in p53 deficient cancer cells exposed to metabolic stress. This effect was mediated by the requirement of p53 deficient cancer cells to synthesise ubiquinone (coenzyme Q10) to maintain TCA cycle activity, respiration and the production of pyrimidine nucleotides. Our study has revealed a novel link between isoprenoid synthesis by the mevalonate pathway and the electron transport function of ubiquinone, which is required for nucleotide biosynthesis. As a consequence, maintaining mevalonate pathway activity is essential for p53 deficient cancer cells to proliferate and survive under the metabolic constraints of the tumor microenvironment.
2019-11-15 | GSE124189 | GEO
Project description:Simvastatin has been widely used for treatment of hypercholesterolemia due to its ability to inhibit HMG-CoA reductase, the rate limiting enzyme of de novo cholesterol synthesis via mevalonate pathway. Its inhibitory action causes also depletion of pathway intermediates, farnesyl pyrophosphate (FPP) and geranyl-geranyl pyrophosphate (GGPP), which are inevitable for proper targeting of small GTPases (e.g. Ras proteins) to their site of action. We profiled by array the gene expression of MIA PaCa-2 cells treated with simvastatin, FPP, GGPP and their combinations. The inhibitory effect of statins on GFP-K-Ras protein trafficking were partially prevented by addition of the mevalonate pathway intermediates. We conclude that the anticancer effect of simvastatin is to a large extent mediated through isoprenoid intermediates of the mevalonate pathway.
2018-03-31 | E-MTAB-3263 | biostudies-arrayexpress
Project description:Cardiac metabolism plays a crucial role in producing sufficient energy to sustain cardiac function. However, the role of metabolism in different aspects of cardiomyocyte regeneration remains unclear. Working with the adult zebrafish heart regeneration model, we first find an increase in the levels of mRNAs encoding enzymes regulating glucose and pyruvate metabolism, including pyruvate kinase M1/2 (Pkm) and pyruvate dehydrogenase kinases (Pdks), especially in tissues bordering the damaged area. We further find that impaired glycolysis decreases the number of proliferating cardiomyocytes following injury. These observations are supported by analyses using loss-of-function models for the metabolic regulators Pkma2 and peroxisome proliferator-activated receptor gamma coactivator 1 alpha. Cardiomyocyte-specific loss- and gain-of-function manipulations of pyruvate metabolism using Pdk3 as well as a catalytic subunit of the pyruvate dehydrogenase complex (PDC) reveal its importance in cardiomyocyte dedifferentiation and proliferation after injury. Furthermore, we find that PDK activity can modulate cell cycle progression and protrusive activity in mammalian cardiomyocytes in culture. Our findings reveal new roles for cardiac metabolism and the PDK-PDC axis in cardiomyocyte behavior following cardiac injury.
2020-07-13 | PXD016235 | Pride
Project description:Some cell type-specific gene expression is maintained in the maturation of cardiomyocytes, where DNA hypomethylation of gene body regions of a set of specific genes. We used microarrays to detail the global gene expression program underlying the maintenance of cardiomyocyte function and maturation and compared it with DNA methylation status. Cardiomyocytes and cardiac fibroblasts were carefully isolated from neonatal and adult hearts and used fresh for the analysis.
2012-07-05 | E-GEOD-39039 | biostudies-arrayexpress
Project description:hiPS derived cardiomycoytes treated with 4 cell cycle genes to induce cardiomyocyte proliferation and sorted for the different cell cycle stages according to FUCCI dye
2018-02-13 | GSE97728 | GEO