AUTHOR=Ma Li , Zhou Na , Zou Rongjun , Shi Wanting , Luo Yuanyuan , Du Na , Zhong Jing , Zhao Xiaodong , Chen Xinxin , Xia Huimin , Wu Yueheng 

TITLE=Single-Cell RNA Sequencing and Quantitative Proteomics Analysis Elucidate Marker Genes and Molecular Mechanisms in Hypoplastic Left Heart Patients With Heart Failure

JOURNAL=Frontiers in Cell and Developmental Biology

VOLUME=9

YEAR=2021

URL=https://www.frontiersin.org/journals/cell-and-developmental-biology/articles/10.3389/fcell.2021.617853

DOI=10.3389/fcell.2021.617853

ISSN=2296-634X

ABSTRACT=<sec><title>Objective</title><p>To probe markers and molecular mechanisms of the hypoplastic left heart (HLH) by single-cell RNA sequencing (scRNA-seq) and quantitative proteomics analysis.</p></sec><sec><title>Methods</title><p>Following data preprocessing, scRNA-seq data of pluripotent stem cell (iPSC)-derived cardiomyocytes from one HLH patient and one control were analyzed by the Seurat package in R. Cell clusters were characterized, which was followed by a <italic>pseudotime</italic> analysis. Markers in the <italic>pseudotime</italic> analysis were utilized for functional enrichment analysis. Quantitative proteomics analysis was based on peripheral blood samples from HLH patients without heart failure (HLH-NHF), HLH patients with heart failure (HLH-HF), and healthy controls. Hub genes were identified by the intersection of <italic>pseudotime</italic> markers and differentially expressed proteins (DE-proteins), which were validated in the GSE77798 dataset, RT-qPCR, and western blot.</p></sec><sec><title>Results</title><p>Cardiomyocytes derived from iPSCs were clustered into mesenchymal stem cells, myocardium, and fibroblast cells. <italic>Pseudotime</italic> analysis revealed their differentiation trajectory. Markers in the three <italic>pseudotime</italic> clusters were significantly associated with distinct biological processes and pathways. Finally, three hub genes (MMP2, B2M, and COL5A1) were identified, which were highly expressed in the left (LV) and right (RV) ventricles of HLH patients compared with controls. Furthermore, higher expression levels were detected in HLH patients with or without HF than in controls.</p></sec><sec><title>Conclusion</title><p>Our findings elucidate marker genes and molecular mechanisms of HLH, deepening the understanding of the pathogenesis of HLH.</p></sec>