■慢性血栓栓塞性肺动脉高压(CTEPH)是一种严重的肺血管疾病,其特征是肺动脉中残留的血栓和远端肺微血管重塑。CTEPH的发病机制尚不清楚,但是很多因素比如炎症,豁免权,可能涉及凝血和血管生成。单核细胞是重要的免疫细胞,可分化为巨噬细胞和树突状细胞,在血栓形成中起重要作用。然而,分布,CTEPH患者单核细胞亚群的基因表达谱和分化轨迹尚未得到系统研究。本研究旨在利用单细胞测序技术揭示CTEPH患者单核细胞的特性和功能,为CTEPH的诊断和治疗提供新的见解。
■进行单细胞RNA测序(scRNA-seq)以分析来自健康对照的外周血单核细胞(PBMC)的转录组特征,CTEPH患者和肺动脉内膜切除术(PEA)后CTEPH患者的组织。通过中心静脉导管反复注射自体血栓,建立CTEPH大鼠慢性肺栓塞模型,流式细胞术检测CTEPH患者和CTEPH大鼠模型中单核细胞亚群的比例变化。我们还通过免疫荧光染色观察了血栓组织中巨噬细胞亚群的浸润程度及其与外周血单核细胞亚群的分化关系。
■结果显示,CTEPH患者外周血单核细胞亚群发生显著变化,尤其是CD16+单核细胞亚群的比例增加。这个单核细胞亚群在转录组水平上具有独特的功能特征,涉及细胞粘附等过程,T细胞活化,凝血反应和血小板活化,在肺动脉血栓形成和肺动脉内膜重构中起重要作用。此外,我们还发现CTEPH患者肺内膜切除组织中的巨噬细胞亚群表现出促炎症和脂质代谢重编程特征,这可能与肺动脉血栓的持续和不溶性以及肺动脉高压的发展有关。最后,我们还观察到CTEPH患者外周血中的CD16单核细胞亚群可能被募集到肺动脉内膜组织并分化为高表达IL-1β的巨噬细胞亚群,参与疾病进展。
■CD16+单核细胞亚群在CTEPH患者中有显著的基因表达变化,与血小板活化有关,凝血反应和炎症反应。并且我们还发现这些细胞可以迁移到血栓并分化为巨噬细胞,高表达IL-1β参与CTEPH疾病的进展。我们认为CD16+单核细胞是CTEPH的重要参与者和潜在的治疗靶点。
UNASSIGNED: Chronic thromboembolic pulmonary hypertension (CTEPH) is a serious pulmonary vascular disease characterized by residual thrombi in the pulmonary arteries and distal pulmonary microvascular remodeling. The pathogenesis of CTEPH remains unclear, but many factors such as inflammation, immunity, coagulation and angiogenesis may be involved. Monocytes are important immune cells that can differentiate into macrophages and dendritic cells and play an important role in thrombus formation. However, the distribution, gene expression profile and differentiation trajectory of monocyte subsets in CTEPH patients have not been systematically studied. This study aims to reveal the characteristics and functions of monocytes in CTEPH patients using single-cell sequencing technology, and to provide new insights for the diagnosis and treatment of CTEPH.
UNASSIGNED: Single-cell RNA sequencing (scRNA-seq) were performed to analyze the transcriptomic features of peripheral blood mononuclear cells (PBMCs) from healthy controls, CTEPH patients and the tissues from CTEPH patients after the pulmonary endarterectomy (PEA). We established a CTEPH rat model with chronic pulmonary embolism caused by repeated injection of autologous thrombi through a central venous catheter, and used flow cytometry to detect the proportion changes of monocyte subsets in CTEPH patients and CTEPH rat model. We also observed the infiltration degree of macrophage subsets in thrombus tissue and their differentiation relationship with peripheral blood monocyte subsets by immunofluorescence staining.
UNASSIGNED: The results showed that the monocyte subsets in peripheral blood of CTEPH patients changed significantly, especially the proportion of CD16+ monocyte subset increased. This monocyte subset had unique functional features at the transcriptomic level, involving processes such as cell adhesion, T cell activation, coagulation response and platelet activation, which may play an important role in pulmonary artery thrombus formation and pulmonary artery intimal remodeling. In addition, we also found that the macrophage subsets in pulmonary endarterectomy tissue of CTEPH patients showed pro-inflammatory and lipid metabolism reprogramming features, which may be related to the persistence and insolubility of pulmonary artery thrombi and the development of pulmonary hypertension. Finally, we also observed that CD16+ monocyte subset in peripheral blood of CTEPH patients may be recruited to pulmonary artery intimal tissue and differentiate into macrophage subset with high expression of IL-1β, participating in disease progression.
UNASSIGNED: CD16+ monocytes subset had significant gene expression changes in CTEPH patients, related to platelet activation, coagulation response and inflammatory response. And we also found that these cells could migrate to the thrombus and differentiate into macrophages with high expression of IL-1β involved in CTEPH disease progression. We believe that CD16+ monocytes are important participants in CTEPH and potential therapeutic targets.