Open Access

ARTICLE

Single-Nucleus RNA Sequencing Reveals Cardiac Macrophage Landscape in Hypoplastic Left Heart Syndrome

by Xiaozhuo Xu^1,2, Yilin Huang^1,2, Xu Han^2,*

1 First Clinical Medical College, Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing, 210008, China
2 Department of Geriatrics, Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing, 210029, China

* Corresponding Author: Xu Han. Email:

Congenital Heart Disease 2024, 19(2), 233-246. https://doi.org/10.32604/chd.2024.050231

Received 31 January 2024; Accepted 16 April 2024; Issue published 16 May 2024

Abstract

Background: Hypoplastic left heart syndrome (HLHS) is one of the most challenging congenital heart diseases in clinical treatment. In cardiac tissues, resident macrophages fulfill critical functions in maintaining a stable cardiac state and have strong regenerative capacity and organ specificity. However, the molecular mechanisms of macrophages in HLHS remained unclear. Methods: Single-nucleus RNA sequencing (snRNA-seq) data of HLHS and healthy control (donors) samples obtained from the Gene Expression Omnibus (GEO) database were normalized and clustered using the Seurat package. The “FindMarkers” function was used to screen differentially expressed genes (DEGs) between the HLHS and donor groups and to analyze the functional enrichment of the set of genes of interest. Finally, cell-cell communication, pseudotime, and single-cell regulatory network inference and clustering (SCENIC) analyses were used to study the mechanisms of macrophages in HLHS. Results: Based on the snRNA-seq data of HLHS and donors, we identified a total of 9 cell clusters, among which the proportion of macrophages was significantly less in the HLHS group than in the control group. Subdivision of macrophage subpopulations (Macrophages 1, 2, and 3) showed that Macrophages 1 was mainly involved in nervous system development, angiogenesis, and apoptotic processes. In addition, analysis of communication between Macrophages 1 and cardiomyocytes revealed that ligand-acceptor pairs such as GAS6/AXL, IL6, IGF1, THY1, and L1CAM were present only in the donor group. Finally, pesudotime and SCENIC analyses demonstrated that FOXO3 and ELF2 played a critical role for Macrophages 1 to maintain cardiac function in patients with HLHS. Conclusion: Our study improved the current understanding of the molecular mechanisms of macrophage development in HLHS, showing that manipulating the regulatory role of macrophages in the heart can be a novel treatment for HLHS.

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Supplementary Material

Supplementary Material File

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