Single cell gene expresison profiling in human embryonic stem cell derived cardiomyocytes
Ontology highlight
ABSTRACT:
ORGANISM(S): Homo sapiens
PROVIDER: GSE157175 | GEO | 2021/03/01
REPOSITORIES: GEO
Similar Datasets
Project description:Myocardial infarction is a major cause of morbidity and mortality worldwide. The limited ability of the surviving cardiac cells to proliferate following an ischemic attack renders the damaged heart susceptible to unfavorable remodeling processes and heart failure. Currently, pharmaceutical and implantable device management of heart failure seek only to preserve existing viable myocardium after an ischemic attack, and thus merely slows the progression of cardiac dysfunction. Ultimately, heart transplantation is the only viable treatment option for end-stage heart failure patients. To “regenerate” the heart and not only preserve cardiac function but also recover lost or diseased muscle, stem cell therapy has emerged as a promising therapy for heart disease because it can provide a virtually unlimited source of cardiomyocytes, endothelial cells, and other differentiated cell types. The hope is to use these cells to replace diseased myocardium that would otherwise progress to outright failure and regenerate the heart to its former, healthy self. Recently, human embryonic stem cells (hESCs) have generated much interest because of their capacity for self-renewal and pluripotency. In practical terms, hESCs can be cultured indefinitely ex vivo, and can differentiate into virtually any cell type in the adult body. hESCs are thus an attractive source for the derivation of large numbers of cells to be used in various tissue repair and cell replacement therapies. However, upon transplantation into living organisms, undifferentiated hESCs can spontaneously differentiate into rapidly proliferating teratomas, which are disordered amalgams of all three germs layers. Nevertheless, under the appropriate conditions, ex vivo hESCs can be directed to differentiate into beating cardiomyocytes via an embryoid body (EB) intermediate. Subsequently, the cardiomyocyte sub-population is enriched several-fold using discontinuous density gradient separation. Therefore, coaxing hESCs into cardiomyocytes for therapeutic applications is an innovative and feasible strategy that can minimize the risk of cellular misbehavior and teratoma formation. In order to define at a molecular level the changes occurring at each stage of hESC differentiation into cardiomyocytes, we performed transcriptional profiling of the cells using whole human genome microarrays. This allowed us to examine the activation of specific genes as well as broader developmental processes during the progression from hESC to fetal cardiomyocyte, and to identify novel genes that are potentially important in mediating differentiation and development as well as potential novel markers of each stage. In the future, such genes may prove vital in efforts to more closely direct and assess differentiation of potential therapeutic pre-cardiomyocytes or cardiomyocytes in the repair of injured cardiac tissues. In our microarray analysis, we observed high expression of pluripotency-related genes involved in the core hESC regulatory circuitry, including OCT4, SOX2, and NANOG, as well as CRYPTO 1 and 3, LCK, and HESX1. Differentiation into beating EBs was accompanied by mesodermal differentiation and dramatic activation of TWIST1, TBX5, and MEOX transcription, as well as the very clear induction of nearly all of the early cardiogenic genes, including FOXC1, ISL2, HAND1, GATA4, 5, and 6, FOXH1, and MEF2C. While it is clear that other developmental lineages are still present in the EB population, it is also clear from the high levels of cardiac gene expression that this population is significantly enriched for the cardiac lineage even at an early stage. The transcriptional analysis of the final differentiation and selection of the hESC-derived CMs indicates that this enrichment continues, with the CM population expressing differentiated cardiomyocyte genes at levels similar to our more advanced FH cells. Importantly, because of the cell type heterogeneity in the fetal heart, we specifically isolated cardiomyocytes from the fetal left ventricles for microarray analysis. In summary, hESC-CMs hold potential promise for treatment of cardiovascular disease. The molecular processes that control stem cell pluripotency, differentiation, and proliferation are complex, justifying the need for a broad investigation that integrates systems biological tools for transcriptome analysis. We found that the enriched hESC-CMs expresses cardiomyocyte genes at levels similar to 20-week fetal heart cells, making this population a good source of potential replacement cells in the in vivo setting.
2008-12-06 | GSE13834 | GEO
Project description:hPSC-CMs resemble immature embryonic or fetal CMs rather than mature adult CMs, and have limitations in disease modeling and pharmacological studies. Therefore, hPSCs-derived mature and ventricular CMs are required for more accurate in vitro modeling of adult-onset cardiac disease and drug discovery. FGF4+AA-treated hESC-CMs robustly released acute myocardial infarction (AMI) biomarkers (cTnI, CK-MB, and myoglobin) into culture medium in response to hypoxic injury. Hypoxia-responsive genes related to cellular responses to glycolytic processes, oxygen levels, HIF-1 signaling, apoptotic processes, and regulation of cell death including potential cardiac biomarkers proved in clinical studies in the diagnosis and prognosis of coronary artery diseases were induced in FGF4+AA-treated hESC-CMs in response to hypoxia based on transcriptome analyses.
2021-04-11 | GSE171824 | GEO
Project description:Human embryonic stem cell-derived cardiomyocytes (hESC-CMs) and human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) grown for 35 days on laminin-coated coverslips, a stiff matrix, were enzymatically dissociated, replated on laminin-coated coverslips and grown for further 2 days. Gene expression was compared with controls grown for 37 days on laminin-coated coverslips. Total RNA was isolated and gene expression was analyzed by RNA-sequencing (RNA-seq) on an Illumina NextSeq 550 sequencer using a High Output Flowcell for single reads (20024906; Illumina). Gene enrichment analysis based on RNA-Seq data of hESC-CMs replated for 2 days as compared with 37 days old controls was performed by using the comprehensive gene set enrichment analysis tool Enrichr for pathway analysis with KEGG Pathways 2019 Human, as well as for Gene Ontology (GO) analysis with GO Cellular Component 2018, GO Molecular Function 2018, and GO Biological Process 2018. Gene enrichment was also analyzed by Ingenuity Pathway analysis (IPA, Qiagen), especially Canonical Pathways analysis. Gene enrichment analysis revealed changes in the gene expression profile, especially of mechanosensation/-transduction-related genes and pathways in replated hESC-CMs.
2023-08-17 | GSE176154 | GEO
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
Project description:Human embryonic stem cell-reporter line hESC-NKX2.5(eGFP/w) were differentiated to cardiomyocytes (CMs) by utilizing the spin embryoid body method. During differentiation the cells were treated with DMSO or retinoic acid (RA) from day 4-7. At day 31, cells were sorted based on GFP prior to RNA isolation. The results of this microarray demonstrate that CMs treated with RA during differentiation exhibit atrial-like gene expression profile, while DMSO-treated cells show ventricular-like gene profile. CMs treated with DMSO or RA during differentiation were sorted for GFP and analyzed for differential gene expression.
2015-01-01 | E-GEOD-61154 | biostudies-arrayexpress
Project description: Not available
| S-EPMC5494509 | biostudies-literature
Project description:To understand the exosomal secretome of ESC-CMs and iPSC-CMs, samples were analyzed using the Human LncRNA Array v4.0 (8 x 60K, Arraystar)
2017-07-19 | GSE100218 | GEO
Project description:To dissect complex regulatory mechanisms on cardiac development, we performed the time-series gene expression experiments of human embryonic stem cells (ESCs) differentiated to cardiomyocytes (CMs). The RNA-sequencing at 4 stages of human to CM maturation, representing timepoints with days 15, 30, 45 and 60 of ESC-CMs. The early CMs from differentiation days (D) 0-15 are commonly considered to be immature and embryonic-like. We define late CMs after D15. During this phase, the cells stop dividing and undergo some degree of maturation. This study established distinct dynamic expression patterns of mRNAs/genes and identified several subsets of up- or down-regulated genes with similar expression patterns during CM differentiation and maturation.
2023-08-07 | GSE239918 | GEO
Project description:Human pluripotent stem cell-derived cardiomyocytes (CMs) are a promising tool for cardiac cell therapy. To optimize graft cells for cardiac reconstruction, we compared the engraftment efficiency of intramyocardially-injected undifferentiated-induced pluripotent stem cells (iPSCs), day4 mesodermal cells, and day8, day20, and day30 purified iPSC-CMs after initial differentiation by tracing the engraftment ratio (ER) using in vivo bioluminescence imaging. This analysis revealed the ER of day20 CMs was significantly higher compared to other cells. Transplantation of day20 CMs into the infarcted hearts of immunodeficient mice showed significant functional improvement. Moreover, the imaging signal and ratio of Ki67-positive CMs at 3 months post injection indicated engrafted CMs proliferated in the host heart. Although this graft growth reached a plateau at 3 months, histological analysis confirmed progressive maturation from 3 to 6 months. These results suggested that day20 CMs had very high engraftment, proliferation, and therapeutic potential in host mouse hearts. Differentiated cells, N=10 Undifferentiated pluripotent stem cells, N=1 Heart samples, N=6
2016-07-03 | E-GEOD-60634 | biostudies-arrayexpress
Project description:Analysis of differential gene expression in Human endometrial endothelial cells (HEECs) incubated with CMS derived from human endoemtrial stromal cells (HESCs) treated by LAPCs. We tested the hypothesis that paracrine factors sectreted from HESCs treated LAPCs, etonogestrol (ETO) or medorxyprogesterone acetate (M) influence several common genes associated with survival in HEECs. Thefore, whole genome analyses were performed in HEECs treated with HESC derived CMS obtained from vehicle (estrodial (E) as control) or progesterone (P) or ETO or M incubations under hypoxia (HX) and normoxia (NX). Results provide important information of several common and unique gene expression profiles in cultured HEECs treated with HESC CMS from P or ETO or M incubations. Among M and ETO responsive genes, up- or down-regulated survival related common genes were determined and used further confirmation and in vitro functional analyses. Total RNA (n=3) obtained from cultured HEECs incubated 6h with vehicle- or P4- or ETO- or MPA- treated NX or HX conditioned HESC-derived CMS.
2014-03-08 | E-GEOD-55691 | biostudies-arrayexpress