Single-cell RNA sequencing reveals a mechanism underlying the susceptibility of the left atrial appendage to intracardiac thrombogenesis during atrial fibrillation

Jie Yang; Hu Tan; Mengjia Sun; Renzheng Chen; Zhao Jian; Yuanbin Song; Jihang Zhang; Shizhu Bian; Bo Zhang; Yi Zhang; Xubin Gao; Zhen Chen; Boji Wu; Xiaowei Ye; Hailin Lv; Zhen Liu; Lan Huang

doi:10.1002/ctm2.1297

Single-cell RNA sequencing reveals a mechanism underlying the susceptibility of the left atrial appendage to intracardiac thrombogenesis during atrial fibrillation

Clin Transl Med. 2023 Jun;13(6):e1297. doi: 10.1002/ctm2.1297.

Authors

Jie Yang^{1

2}, Hu Tan^{1

2}, Mengjia Sun^{1

2}, Renzheng Chen^{1

2}, Zhao Jian³, Yuanbin Song^{1

2}, Jihang Zhang^{1

2}, Shizhu Bian^{1

2}, Bo Zhang^{1

2}, Yi Zhang^{1

2}, Xubin Gao^{1

2}, Zhen Chen^{1

2}, Boji Wu^{1

2}, Xiaowei Ye^{1

2}, Hailin Lv^{1

2}, Zhen Liu^{1

2}, Lan Huang^{1

2}

Affiliations

¹ Institute of Cardiovascular Diseases of PLA, the Second Affiliated Hospital, Army Medical University (Third Military Medical University), Chongqing, China.
² Department of Cardiology, the Second Affiliated Hospital, Army Medical University (Third Military Medical University), Chongqing, China.
³ Department of Cardiovascular Surgery, the Second Affiliated Hospital, Army Medical University (Third Military Medical University), Chongqing, China.

Abstract

Background: Atrial fibrillation (AF) is associated with an increased risk of thrombosis of the left atrial appendage (LAA). However, the molecular mechanisms underlying this site-specificity remain poorly understood. Here, we present a comparative single-cell transcriptional profile of paired atrial appendages from patients with AF and illustrate the chamber-specific properties of the main cell types.

Methods: Single-cell RNA sequencing analysis of matched atrial appendage samples from three patients with persistent AF was evaluated by 10× genomics. The AF mice model was created using Tbx5 knockout mice. Validation experiments were performed by glutathione S-transferase pull-down assays, coimmunoprecipitation (Co-IP), cleavage assays and shear stress experiments in vitro.

Results: In LAA, phenotype switching from endothelial cells to fibroblasts and inflammation associated with proinflammatory macrophage infiltration were observed. Importantly, the coagulation cascade is highly enriched in LAA endocardial endothelial cells (EECs), accompanying the up-regulation of a disintegrin and metalloproteinase with thrombospondin motifs 1 (ADAMTS1) and the down-regulation of the tissue factor pathway inhibitor (TFPI) and TFPI2. Similar alterations were verified in an AF mouse model (Tbx5^+/- ) and EECs treated with simulated AF shear stress in vitro. Furthermore, we revealed that the cleavage of both TFPI and TFPI2 based on their interaction with ADAMTS1 would lead to loss of anticoagulant activities of EECs.

Conclusions: This study highlights the decrease in the anticoagulant status of EECs in LAA as a potential mechanism underlying the propensity for thrombosis, which may aid the development of anticoagulation therapeutic approaches targeting functionally distinct cell subsets or molecules during AF.

Keywords: ADAMTS1; TFPI family; atrial fibrillation; endocardial endothelial cells; single-cell RNA-sequencing; thrombogenesis.

MeSH terms

Animals
Anticoagulants / metabolism
Atrial Appendage* / metabolism
Atrial Fibrillation* / complications
Atrial Fibrillation* / genetics
Endothelial Cells / metabolism
Mice
Sequence Analysis, RNA
Thrombosis* / genetics

Substances

Anticoagulants