Abnormal Differentiation and Proliferation of Coronary Arterial Endothelium in Hypoplastic Left Heart Syndrome

Hypoplastic left heart syndrome (HLHS) is a severe form of single ventricle congenital heart disease characterized by an underdevelopment of the left ventricle. Early serial postmortem examinations revealed high rate of coronary artery abnormalities in HLHS fetal hearts, which may impact ventricular development and intra-cardiac hemodynamics, leading to a poor prognosis after surgical palliations. Previous study reported that endothelial cells (ECs) lining the coronary vessels showed DNA damage in the left ventricle of human fetal heart with HLHS, indicating that EC dysfunction may contribute to the coronary abnormalities in HLHS. To investigate the underlying mechanism of HLHS coronary artery abnormalities, we profiled both human fetal heart with an underdeveloped left ventricle (ULV) and ECs differentiated from induced pluripotent stem cells (iPSCs) derived from HLHS patients at single cell resolution. CD144+/NPR3- vascular ECs were selected and further classified as venous EC (NR2F2high), arterial EC (EFNB2high) and late arterial EC (GJA5high) subclusters based on previously reported marker genes. To study the arterial phenotype, we specifically generated iPSC-arterial ECs (AECs, CD34+CDH5+CXCR4+NT5E-/low) derived from three HLHS patients and three age-matched healthy controls to further dissect the phenotype of HLHS-AECs. As compared to normal human heart and control iPSC-ECs respectively, ULV late arterial EC subcluster and HLHS iPSC-EC arterial clusters showed significantly reduced expression of arterial genes GJA5, DLL4, and HEY1. Pathway enrichment analysis based on differentially expressed genes revealed several defects in late AEC cluster from ULV compared to normal human heart, such as impaired endothelial proliferation, development and Notch signaling. HLHS iPSCs exhibited impaired AEC differentiation as evidenced by the significantly reduced CXCR4+NT5E-/low AEC progenitor population. Consistent with human heart transcriptomic data, matured HLHS iPSC-AECs also showed a lower expression of the arterial genes such as GJA5, DLL4, HEY1 compared with control. Additionally, matured HLHS iPSC-AECs showed significantly decreased expression of cell proliferation marker Ki67 and G1/S transition genes (CCND1, CCND2) compared with control, indicating that HLHS iPSC-AECs largely resided in the G0/G1 phase and failed to enter the cell cycle normally. In summary, we found that coronary AECs from HLHS showed impaired arterial development and proliferation. These functional defects in HLHS coronary AECs could contribute to the vascular structure malformation and impaired ventricular development.

Download Full-text

Distention of the Immature Left Ventricle Triggers Development of Endocardial Fibroelastosis: An Animal Model of Endocardial Fibroelastosis Introducing Morphopathological Features of Evolving Fetal Hypoplastic Left Heart Syndrome

BioMed Research International ◽

10.1155/2015/462469 ◽

2015 ◽

Vol 2015 ◽

pp. 1-10 ◽

Cited By ~ 5

Author(s):

Shogo Shimada ◽

Christian Robles ◽

Ben M. W. Illigens ◽

Alejandra M. Casar Berazaluce ◽

Pedro J. del Nido ◽

...

Keyword(s):

Animal Model ◽

Left Ventricle ◽

Hypoplastic Left Heart Syndrome ◽

Left Heart ◽

Endocardial Fibroelastosis ◽

Hypoplastic Left Heart ◽

Fetal Imaging ◽

Adult Rat ◽

Human Fetal Heart ◽

Left Heart Syndrome

Background.Endocardial fibroelastosis (EFE), characterized by a diffuse endocardial thickening through collagen and elastin fibers, develops in the human fetal heart restricting growth of the left ventricle (LV). Recent advances in fetal imaging indicate that EFE development is directly associated with a distended, poorly contractile LV in evolving hypoplastic left heart syndrome (HLHS). In this study, we developed an animal model of EFE by introducing this human fetal LV morphopathology to an immature rat heart.Methods and Results.A neonatal donor heart, in which aortic regurgitation (AR) was created, was heterotopically transplanted into a recipient adult rat. AR successfully induced the LV morphology of evolving HLHS in the transplanted donor hearts, which resulted in the development of significant EFE covering the entire LV cavity within two weeks postoperatively. In contrast, posttransplants with a competent aortic valve displayed unloaded LVs with a trace of EFE.Conclusions.We could show that distention of the immature LV in combination with stagnant flow triggers EFE development in this animal model. This model would serve as a robust tool to develop therapeutic strategies to treat EFE while providing insight into its pathogenesis.

Download Full-text

Abstract 238: Single-Cell Transcriptomic Analysis and Patient-Specific IPSCs Reveal Dysregulated Cell Cycle in Coronary Endothelial Cell in Hypoplastic Left Heart Syndrome

Circulation Research ◽

10.1161/res.127.suppl_1.238 ◽

2020 ◽

Vol 127 (Suppl_1) ◽

Author(s):

Mingxia Gu ◽

Yifei Miao ◽

Xin Zhou ◽

Lei Tian ◽

Marcy Martin ◽

...

Keyword(s):

Cell Cycle ◽

Endothelial Cell ◽

Left Ventricle ◽

Single Cell ◽

Hypoplastic Left Heart Syndrome ◽

Human Heart ◽

G1 Phase ◽

Left Heart ◽

Hypoplastic Left Heart ◽

Left Heart Syndrome

Hypoplastic left heart syndrome (HLHS) is a single ventricle congenital heart disease that results in severe underdevelopment of the left ventricle, mitral valve, aortic valve, and ascending aorta. Early serial postmortem examinations also revealed a high rate of coronary anomalies in HLHS, which included multiple ventriculo-coronary arterial connections as well as thick-walled and kinked coronary arteries. A previous study showed that fetal hypoplastic left hearts had a reduced endothelial cell (EC) population and lower capillary density compared with normal hearts. However, the mechanism underlying coronary abnormalities associated with HLHS remains unknown. Thus, we generated induced pluripotent stem cells derived ECs (iPSC-ECs) from three HLHS patients and three age-matched controls. Single Cell RNA-Seq (scRNA-seq) profiling identified both endocardial (NPR3 + /CDH5 + ) and coronary endothelial populations (APLN + /CDH5 + ) from the heterogeneous iPSC-ECs. Intriguingly, a subcluster of the coronary endothelial cells (CECs) with cell cycle arrest was specifically enriched in HLHS patients. Further cell cycle analysis showed that 30.6% of the HLHS cells were trapped in the G1 phase, while the majority of the control CECs entered cell cycle normally. Additionally, the cell cycle differences between control and HLHS was only seen in CECs, not in the endocardial population. To verify our transcriptomic analysis, we applied negative cell sorting (NPR3 - /CDH5 + ) on iPSC-ECs to purify CECs (iCECs) and confirmed that HLHS iCECs showed profound reduction of cell cycle/proliferative genes ( KI67, PCNA, CCNA2, CCNB1 ) and abnormal induction of CCND2 , which is the hallmark of G1 phase. BrdU assays also indicated suppressed proliferation in HLHS iCECs. Furthermore, we profiled the transcriptome from a human heart with an underdevelopment left ventricle (ULV) at single cell resolution. When compared to the normal human heart, pathway enrichment analysis of differentially expressed genes in ULV hearts demonstrated reduced cell proliferation in the CEC subpopulation. Here, we identified that CECs from HLHS patients exerted proliferative defects that can potentially impede the development of vascular/capillary structure and cause related functional deficiencies. Reformation of the coronary defect provides a promising therapeutic strategy to prevent HLHS deterioration.

Download Full-text

Coronary artery abnormalities and right ventricular histology in hypoplastic left heart syndrome

Journal of the American College of Cardiology ◽

10.1016/0735-1097(92)90101-r ◽

1992 ◽

Vol 20 (2) ◽

pp. 350-358 ◽

Cited By ~ 73

Author(s):

Jeanne Marie Baffa ◽

Sheng-Liang Chen ◽

Marta E. Guttenberg ◽

William I. Norwood ◽

Paul M. Weinberg

Keyword(s):

Coronary Artery ◽

Right Ventricular ◽

Hypoplastic Left Heart Syndrome ◽

Left Heart ◽

Hypoplastic Left Heart ◽

Coronary Artery Abnormalities ◽

Left Heart Syndrome

Download Full-text

Hypoplastic left heart syndrome associated with left ventricle-coronary artery-pulmonary artery fistula: prenatal diagnosis and histological correlation

Ultrasound in Obstetrics and Gynecology ◽

10.1002/uog.1932 ◽

2005 ◽

Vol 26 (1) ◽

pp. 78-80 ◽

Cited By ~ 8

Author(s):

D. Paladini ◽

G. Caruso ◽

G. Sglavo ◽

C. Lapadula ◽

G. Parisi ◽

...

Keyword(s):

Prenatal Diagnosis ◽

Coronary Artery ◽

Left Ventricle ◽

Pulmonary Artery ◽

Hypoplastic Left Heart Syndrome ◽

Left Heart ◽

Hypoplastic Left Heart ◽

Pulmonary Artery Fistula ◽

Left Heart Syndrome

Download Full-text

Total anomalous pulmonary venous connection mimicking hypoplastic left heart syndrome

Cardiology in the Young ◽

10.1017/s1047951121001670 ◽

2021 ◽

pp. 1-3

Author(s):

Ryohei Matsuoka ◽

Jun Muneuchi ◽

Yoshie Ochiai

Keyword(s):

Left Ventricle ◽

Systemic Circulation ◽

Z Score ◽

Left Heart ◽

Hypoplastic Left Heart ◽

Transverse Arch ◽

Anomalous Pulmonary Vein ◽

Pulmonary Arterial ◽

Left Heart Syndrome ◽

Anomalous Pulmonary Venous Connection

Abstract A newborn with supracardiac total anomalous pulmonary venous connection vein presented the small left ventricle with z score of −7.5, retrograde blood supply in the transverse arch, and the dutcus-dependent systemic circulation. The patient underwent the repair of the anomalous pulmonary vein and bilateral pulmonary arterial banding soon after the birth and then transcatheter pulmonary arterial debanding at the age of 10 months because of an appropriate growth of the left ventricle.

Download Full-text