Dataset Information

Gene expression profile at single cell level of lung from TRAF5 knockout mouse with Systemic Lupus Erythematosus-associated Pulmonary Arterial Hypertension (SLE-PAH)

ABSTRACT: Pulmonary arterial hypertension (PAH) is a severe complication that significantly impacts the prognosis of patients with systemic lupus erythematosus (SLE). SLE-PAH has high mortality and morbidity but poorly understood pathogenesis. Our goal is to identify novel susceptibility genes associated with SLE-PAH by performing genetic analyses on large cohorts and validating the functional significance. On the genetic level, Whole Exome Sequencing (WES) and Sanger sequencing were performed on the discovery and validation cohorts to identify susceptibility genes associated with SLE-PAH; At the transcriptional level, the expression of susceptibility gene was examined by RNA-seq and RT-qPCR. TRAF5 knockout mice were compared with control after induction of the previously established SLE-PAH model, and the protective effects of TRAF5 overexpression were evaluated. Single-cell RNA sequencing (scRNA-seq) analyses were conducted on lung tissues to investigate pulmonary cellular population. In vitro interventional studies involving TRAF5 knockdown in pulmonary arterial endothelial cells (PAECs) were performed to further investigate its functions. Supporting data from both the discovery and validation cohorts revealed a total of 7 deleterious variants on TRAF5 in 8 cases of SLE-PAH (n=226), with none found in the health control group (HC, n=934). There was decreased TRAF5 expression at the transcriptional level (n=120) compared to SLE-nonPAH (n=116) and HC (n=20). Mice with TRAF5 knockout exhibited more severe pulmonary hypertension, right ventricular hypertrophy, and early mortality. scRNA-seq analyses of lung tissue in knockout mice revealed significantly decreased TRAF5 expression and increased apoptosis in arterial ECs, with distinct pathways identified. In vitro interventional studies confirmed that TRAF5 deficiency caused dysfunction in PAECs, regulating the BMP/TGF-β pathway. Endothelial-targeted overexpression of TRAF5 attenuated PH in model mice.

ORGANISM(S): Mus musculus

PROVIDER: GSE276751 | GEO | 2025/08/31

REPOSITORIES: GEO

ACCESS DATA

Dataset's files

Source:

			Action	DRS
		Other

Items per page:

1 - 1 of 1

Similar Datasets

Project description:Dysfunction of pulmonary arterial endothelial cells (PAECs) is associated with the development and progression of vascular pathology. However, it remains unknown how pulmonary hypertension (PH) affects cellular composition and transcriptomic profile of pulmonary endothelium. Here, we have undertaken a single-cell, compartment specific approach to characterise alterations in PAECs associated with two different types of PH, i.e., pulmonary arterial hypertension (PAH) and pulmonary hypertension associated with pulmonary fibrosis (PHPF). Our unbiased analysis showed that endothelium of medium / small caliber pulmonary arteries is composed of three subsets of endothelial cells (ECs). The analysis of healthy and PH endothelium revealed that the three populations are persistently represented in remodelled arteries. Additionally, an exploratory analysis of human aorta (AO) and coronary arteries (CA) endothelium revealed that, although similar gene expression patterns were noticeable, PAECs subpopulations proportions differs significantly from pulmonary arteries (PA) endothelium. To address whether EC heterogeneity is a prime feature of human endothelium, we also performed a similar analysis in a murine model of hypoxia, revealing that similar EC populations were evident in this animal model. Comparative analysis of EC subpopulations in healthy and PH EC identified a common genetic deregulation accompanying vascular remodelling. Even though murine EC displayed some similarities with human EC subpopulations, the intense re-programming associated with hypoxia associated vascular remodelling displayed significant differences compared to the human disease. Finally, in depth comparative analysis of PAH and PHPF EC highlighted the development of disease-specific transcriptomic alterations in the three populations. Therefore, characterisation of transcriptomic differences in the endothelial bed of PAH and PHPF patients can facilitate identification of novel, disease-specific therapeutic targets.

Project description:Background: Hemodynamic forces exert a profound influence on endothelial cell signaling and, when abnormal, contribute centrally to human vascular disease. Pulmonary arterial hypertension (PAH) is characterized by both hemodynamic derangement and pulmonary arterial endothelial cell (PAEC) dysfunction. Despite importance in disease initiation and progression, the combined effects of shear and pressure forces on PAEC biology remain incompletely understood, particularly in the context of PAH. Methods: PAECs obtained at explant from controls and from patients with idiopathic or congenital heart disease-associated PAH (CHD-PAH) were cultured in a custom resistor-coupled microfluidic platform and exposed to static, low (3 dyne/cm²), or high (20 dyne/cm²) shear stress under either low or elevated (60 mmHg) pressure. After 24 hours, we assessed cellular morphology and performed systems-level transcriptomic analysis via bulk RNA sequencing, incorporating analyses of PAH subtype and donor sex. Results: PAECs (n=18 donors) aligned with flow under high, but not low, shear, and alignment was not significantly altered by disease state or pressure. Shear stress fundamentally reorganized the PAEC transcriptome and the “dose-response” to increasing shear differed across biological pathways in six statistically significant patterns. Increasing shear led to divergence in transcription between control and PAH cells, particularly in pathways involved in immune activation, stress signaling, and vascular remodeling, with subtype differences also observed. Pressure alone had modest effects on transcription, though CHD-PAH PAECs especially displayed pressure-induced stress and inflammatory signaling. We identified sexual dimorphism in the endothelial shear response, noting male cells under shear enriched for pathways involved in proliferation and inflammation and female cells enriched for lipid metabolism and stress responses. Conclusions: Shear and pressure forces profoundly influence PAEC transcription, with responses shaped by disease state, PAH subtype, and sex. These findings highlight the need for further investigation into mechanosensitive pathways in PAH as potential targets for novel therapies.

Dataset Information

Gene expression profile at single cell level of lung from TRAF5 knockout mouse with Systemic Lupus Erythematosus-associated Pulmonary Arterial Hypertension (SLE-PAH)

Dataset's files

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets