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Single-Cell RNA-Seq Analysis Reveals Macrophage Involved in Pathogenesis of Human sporadic type A aortic dissection

ABSTRACT: Macrophages play an important role in the progression of sporadic acute type A aortic dissection (ATAAD). The aim of this study was to characterize the cellular heterogeneity of macrophages in AD tissues by scRNA-seq. Ascending aortic wall tissue from 6 ATAAD patients and 3 heart transplant donors were assessed by scRNA-seq, then analyzed and validated by various bioinformatics algo-rithms and histopathology experiments. The results revealed the proportion of macrophages in AD tissues (24.51%) was significantly higher than that in normal tissues (13.69%). Among the 6 macro-phage subclusters, pro-inflammatory macrophages accounted for 14.96% in the AD group, and 0.18% in the normal group. Chemokine and inflammation related genes (CCL2, CCL20, S100A8, and S100A9) were expressed more intensively in macrophages in AD tissue than that in the normal tissues. Ad-ditionally, intercellular communication analysis and transcription factor analysis indicated activation of inflammation and degradation of the extracellular matrix in AD tissues. Finally, immunohisto-chemistry, immunofluorescence and western blot experiments confirmed the overexpression of macrophage marker genes (CD68 and CD163) and matrix metalloproteinases (MMP9 and MMP2) in AD tissues. Collectively, our study provides a preliminary evaluation of the role of macrophages in AD and potentially aid in the development of therapeutic options in the future.

ORGANISM(S): Homo sapiens

PROVIDER: GSE213740 | GEO | 2022/09/23

REPOSITORIES: GEO

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Project description:Background and aims: Aortic dissection (AD) is a cardiovascular emergency with degeneration of the aortic media. Mounting evidence indicates obstructive sleep apnea (OSA) as an independent risk factor for AD development with unknown mechanisms. This study aims to establish a stable murine model of OSA-related AD (OSA-AD) and uncover the potential changes in gene transcripts in OSA-AD. Materials and methods: ApoE-/- mice were exposed to the chronic intermittent hypoxia (CIH) system combined with Ang II administration to establish the OSA-AD model. Pathological staining was performed to exhibit the physiological structure of the mouse aorta. The SBC mouse ceRNA microarray was used to identify significantly differentially expressed (DE) mRNAs, DE long-non-coding RNAs (DElncRNAs), and DE circular RNAs (DEcircRNAs) in OSA-AD tissues. Subsequently, bioinformatics analysis, including Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genome (KEGG), and protein–protein interaction (PPI) analyses, was performed to evaluate the function of the significantly differentially expressed transcripts (DETs). The hub genes were confirmed using quantitative real-time polymerase chain reaction (qRT-PCR). Results: ApoE-/- mice exposed to CIH and Ang II showed a high ratio of aortic accident (73.33%) and significant aortic diameter dilatation (1.96 ± 0.175 mm). A total of 1,742 mRNAs, 2,625 lncRNAs, and 537 circRNAs were identified as DETs (LogFC ≥ 1.5 or ≤ –1.5, P < 0.05). GO and KEGG analyses demonstrated that the differentially expressed mRNAs (DEmRNAs) were most enriched in cell proliferation, migration, apoptosis, inflammation, and hypoxia-related terms, which are closely related to aortic structural homeostasis. The PPI network contained 609 nodes and 934 connections, the hub genes were highlighted with the CytoHubba plugin and confirmed by qRT-PCR in AD tissues. KEGG pathway analysis revealed that the cis-regulated genes of DElncRNAs and circRNAs-host genes were enriched in aortic structural homeostasis-related pathways. Conclusion: Our findings help establish a de novo OSA-AD animal model using ApoE-/- mice. Many DEmRNAs, DElncRNAs, and DEcircRNAs were screened for the first time in OSA-AD tissues. Our findings provide useful bioinformatics data for understanding the molecular mechanism of OSA-AD and developing potential therapeutic strategies for OSA-AD.

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Single-Cell RNA-Seq Analysis Reveals Macrophage Involved in Pathogenesis of Human sporadic type A aortic dissection

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