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Single-Cell Transcriptomics of Engineered Cardiac Tissues from Patient-Specific Induced Pluripotent Stem Cell-Derived Cardiomyocytes Reveals Abnormal Developmental Trajectory and Intrinsic Contractile Defects in Hypoplastic Right Heart Syndrome

ABSTRACT: Single-Cell Transcriptomics of Engineered Cardiac Tissues from Patient-Specific Induced Pluripotent Stem Cell-Derived Cardiomyocytes Reveals Abnormal Developmental Trajectory and Intrinsic Contractile Defects in Hypoplastic Right Heart Syndrome

PROVIDER: PRJNA660336 | ENA |

REPOSITORIES: ENA

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Single-Cell Transcriptomics of Engineered Cardiac Tissues from Patient-Specific Induced Pluripotent Stem Cell-Derived Cardiomyocytes Reveals Abnormal Developmental Trajectory and Intrinsic Contractile Defects in Hypoplastic Right Heart Syndrome

Project description:Pulmonary atresia with intact ventricular septum (PAIVS) is a subset of hypoplastic right heart syndrome, a form of congenital heart disease (CHD). Genomic studies on PAIVS patients revealed inconclusive genomic variations and polymorphisms and no animal model yet exists for this disease. Hence, we performed single cell RNA-sequencing (scRNA-seq) on PAIVS patient derived human induced pluripotent stem cell cardiomyocytes (hiPSC-CMs) and its bioengineered cardiac tissue constructs (Human cardiac anisotropic sheeet (hCAS) and cardiac tissue strip (hCTS)) to dissect the intrinsic cardiomyocyte defects. Using pseudotime analysis, we identified pro-maturational cues conferred to hiPSC-CMs by hCAS and hCTS and are attenduated in PAIVS. Moerover, we also identified diminished expression of cardiac contractile apparatus genes in PAIVS hiPSC-CMs and hCTS. Our findings reveal intrinsic abnormalities in cardiac contraction and development in PAIVS in the absence of secondary in vivo remodelling.

2020-09-01 | GSE157157 | GEO

Single-Cell Transcriptomics of Engineered Cardiac Tissues From Patient-Specific Induced Pluripotent Stem Cell-Derived Cardiomyocytes Reveals Abnormal Developmental Trajectory and Intrinsic Contractile Defects in Hypoplastic Right Heart Syndrome.

Project description:Background To understand the intrinsic cardiac developmental and functional abnormalities in pulmonary atresia with intact ventricular septum (PAIVS) free from effects secondary to anatomic defects, we performed and compared single-cell transcriptomic and phenotypic analyses of patient- and healthy subject-derived human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) and engineered tissue models. Methods and Results We derived hiPSC lines from 3 patients with PAIVS and 3 healthy subjects and differentiated them into hiPSC-CMs, which were then bioengineered into the human cardiac anisotropic sheet and human cardiac tissue strip custom-designed for electrophysiological and contractile assessments, respectively. Single-cell RNA sequencing (scRNA-seq) of hiPSC-CMs, human cardiac anisotropic sheet, and human cardiac tissue strip was performed to examine the transcriptomic basis for any phenotypic abnormalities using pseudotime and differential expression analyses. Through pseudotime analysis, we demonstrated that bioengineered tissue constructs provide pro-maturational cues to hiPSC-CMs, although the maturation and development were attenuated in PAIVS hiPSC-CMs. Furthermore, reduced contractility and prolonged contractile kinetics were observed with PAIVS human cardiac tissue strips. Consistently, single-cell RNA sequencing of PAIVS human cardiac tissue strips and hiPSC-CMs exhibited diminished expression of cardiac contractile apparatus genes. By contrast, electrophysiological aberrancies were absent in PAIVS human cardiac anisotropic sheets. Conclusions Our findings were the first to reveal intrinsic abnormalities of cardiomyocyte development and function in PAIVS free from secondary effects. We conclude that hiPSC-derived engineered tissues offer a unique method for studying primary cardiac abnormalities and uncovering pathogenic mechanisms that underlie sporadic congenital heart diseases.

| S-EPMC7763394 | biostudies-literature

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Identifying gene expression profiles in hypoplastic left heart syndrome and age-matched normal controls

2011-12-31 | GSE23959 | GEO

Hypoplastic Left Heart Syndrome Single Cell RNA Sequencing

Project description:Single cell transcriptome analysis was performed on one Hypoplastic Left Heart Synrome (HLHS) cell line and one control cell line.

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Fetal hypoplastic lungs have multilineage inflammation that is reversed by amniotic fluid stem cell extracellular vesicle treatment

Project description:We performed single nucleus RNA-seq on rat fetal lungs from: control (normal), hypoplastic (injury), and hypoplastic+ amniotic fluid stem cell extracellular vesicles treated (injury+treatment).

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Single-Cell RNA-Seq and Patient-Specific iPSCs Reveal Endocardial Abnormalities in Hypoplastic Left Heart Syndrome

Project description:Hypoplastic left heart syndrome (HLHS) is one of the most devastating forms of congenital heart defects. Previous studies have only focused on intrinsic defects in the myocardium. However, this does not sufficiently explain the abnormal development of the cardiac valve, septum, and vasculature, which are known to originate from the endocardium. Here, using single-cell RNA profiling, induced pluripotent stem cells, and fetal heart tissue with an underdeveloped left ventricle, we identified a developmentally impaired endocardial cell population in HLHS. The intrinsic endocardial deficits contributed to abnormal endothelial to mesenchymal transition, NOTCH signaling, and extracellular matrix organization, all of which are key factors in valve formation. Consequentially, endocardial abnormalities conferred reduced proliferation and maturation of cardiomyocytes through a disrupted fibronectin-integrin interaction. Several known HLHS de novo mutations all contributed to the abnormal endocardial gene expression through the alteration of promoter/enhancer activities. These mechanistic discoveries provide an alternative angle for early intervention and heart regeneration in HLHS.

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Amniotic fluid stem cell extracellular vesicle therapy improves lung growth and branching morphogenesis in fetal hypoplastic lungs secondary to oligohydramnios

Project description:Oligohydramnios is known to cause fetal pulmonary hypoplasia. Amniotic fluid stem cell extracellular vesicles (AFSC-EVs) have shown beneficial effects on hypoplastic lungs secondary to congenital diaphragmatic hernia. Herein, we investigated, whether AFSC-EVs also foster antenatal lung development in hypoplastic lungs secondary to oligohydramnion. We therefore induced surgically oligohydramnion in rat dams and harvested their fetuses. Controls underwent surgery without amniotic fluid drainage. The hypoplastic lungs were assigned to ex vivo to either medium alone or medium + AFSC-EV as a lung explant. Those samples were harvested 24h later and subjected to RNA extraction for subsequent RNA-seq.

2024-10-17 | GSE247978 | GEO

Identifying gene expression profiles in hypoplastic left heart syndrome and age-matched normal controls

Project description:Affymetrix GeneChip Exon-1.0ST was used to study the differential gene profiles in RV (right ventricle) samples from neonates with HLHS (hypoplastic left heart syndrome) versus RV and LV (left ventricle) samples obtained from age-matched controls. Although few significant changes were observed in the genetic profiles between control LV and control RV, many genes passed the false discovery rate in comparing HLHS-RV to RV and LV control groups, with greater differential profiles noted between HLHS-RV and control RV. Myocardial samples were isolated from the RV of 6 HLHS neonates, diagnosed based upon clinical features including hypoplasia/atresia of the ascending aorta, various degrees of underdevelopment of the aortic valve, mitral valve, and LV cavity, and retrograde flow in the aortic arch as determined by conventional 2-D echocardiography. The mean gestational age at birth of all subjects was 38 weeks (range 36-39) and the mean body weight at surgery of 2.7 kg (range 2.1-3.4 kg) (3 males, 3 females). All subjects were undergoing stage 1 Norwood reconstruction. Children with HLHS and other complex cardiac anomalies entailing non-HLHS single ventricle circulation were excluded from our study. For control samples, RV and LV myocardial tissue was obtained from 5 newborns aged between 1-28 days (mean 18 days; 3 males and 2 females) with normal cardiac anatomy and expired from non-cardiac diseases processes.

2011-12-31 | E-GEOD-23959 | biostudies-arrayexpress

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