Department of Cardio-Pulmonary Circulation, Shanghai Pulmonary Hospital, Tongji University School of Medicine, Shanghai, China (R.Z., S.-G.G., Q.-H.Z., R.J., H.-L.Q., H.-T.L., J.H., L.W.).
Department of Biological Sciences and Technology, College of Biological Science and Medical Engineering, Donghua University, Shanghai, China (R.Z., Y.-L.Z., X.-J.W., J-Y.Z.).
Hypertension. 2023 Aug;80(8):1784-1794. doi: 10.1161/HYPERTENSIONAHA.123.21142. Epub 2023 Jun 14.
Idiopathic pulmonary hypertension (IPAH) is a rare and devastating disease often accompanied by persistent inflammation and immune responses. We aim to provide a reference atlas of neutrophils to facilitate a better understanding of cellular phenotypes and discovery of candidate genes.
Peripheral neutrophils from naive patients with IPAH and matched controls were profiled. Whole-exon sequencing was performed to exclude known genetic mutations before establishing single-cell RNA sequencing. Marker genes were validated by flow cytometry and histology in a separate validation cohort.
Seurat clustering analysis revealed that the landscape of neutrophils encompassed 5 clusters, including 1 progenitor, 1 transition, and 3 functional clusters. The intercorrelated genes in patients with IPAH were mainly enriched in antigen processing presentation and natural killer cell mediated cytotoxicity. We identified and validated differentially upregulated genes, including (matrix metallopeptidase 9), (ISG15 ubiquitin-like modifier), and (C-X-C motif ligand 8). The positive proportions and fluorescence quantification of these genes were significantly increased in CD16 neutrophils in patients with IPAH. The higher proportion of positive MMP9 neutrophils increased mortality risk after adjustment for age and sex. Patients with higher proportions of positive MMP9 neutrophils had worse survival, while the fraction of ISG15- or CXCL8-positive expression neutrophils failed to predict outcome.
Our study yields a comprehensive dataset of the landscape of neutrophils in patients with IPAH. The predictive values of a neutrophil cluster characterized by higher MMP9 expression indicate a functional role for neutrophil-specific matrix metalloproteinases in the pathogenesis of pulmonary arterial hypertension.
特发性肺动脉高压（IPAH）是一种罕见且严重的疾病，常伴有持续的炎症和免疫反应。我们旨在提供中性粒细胞的参考图谱，以帮助更好地理解细胞表型和发现候选基因。
对来自初治 IPAH 患者和匹配对照者的外周血中性粒细胞进行分析。在建立单细胞 RNA 测序之前，进行全外显子测序以排除已知的遗传突变。在另一个验证队列中，通过流式细胞术和组织学验证标记基因。
Seurat 聚类分析显示，中性粒细胞的景观包括 5 个簇，包括 1 个祖细胞、1 个过渡和 3 个功能簇。IPAH 患者的相互关联基因主要富集在抗原加工呈递和自然杀伤细胞介导的细胞毒性中。我们鉴定并验证了差异上调的基因，包括 （基质金属蛋白酶 9）、 （ISG15 泛素样修饰酶）和 （C-X-C 基序配体 8）。这些基因在 IPAH 患者的 CD16 中性粒细胞中的阳性比例和荧光定量均显著增加。经年龄和性别调整后，MMP9 阳性中性粒细胞比例较高的患者死亡率风险增加。MMP9 阳性中性粒细胞比例较高的患者生存预后较差，而 ISG15 或 CXCL8 阳性表达中性粒细胞的比例未能预测预后。
我们的研究提供了 IPAH 患者中性粒细胞景观的全面数据集。以更高 MMP9 表达为特征的中性粒细胞簇的预测值表明中性粒细胞特异性基质金属蛋白酶在肺动脉高压发病机制中的功能作用。
