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Single Cell RNA sequencing of the Monocytes from controls and pulmonary arterial hypertension (PAH) patients

ABSTRACT: We report the application of single cell RNA sequencing on human Peripheral Blood Mononuclear cells. The 12 samples are sequenced as biological replications (6 control and 6 patients), and pooled at the analysis stage. The library was prepared using the Chromium Single Cell 3' v2 and v3 Reagent Kit, and sample processing was under bcl2fastq2/2.20.0 and cellranger/2.2.0. We report a decreased CD14 in classical monocytes among PAH patients, with greater potential of apoptosis for CD14+ monocytes population. This study provide a reference for studying the monocytes related abonormalties in PAH and other inflammatory diseases.

ORGANISM(S): Homo sapiens

PROVIDER: GSE233189 | GEO | 2023/06/01

REPOSITORIES: GEO

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Single Cell RNA sequencing of the Dendritic Cells from controls and pulmonary arterial hypertension (PAH) patients

Project description:We report the application of single cell RNA sequencing on human dendritic cells. By using the negative selecting kit, we enriched theconventional DC from human PBMCs. The seven samples are sequenced as biological replications (4 control and 3 patients), and pooled at the analysis stage. The library was prepared using the Chromium Single Cell 3' v2 Reagent Kit, and sample processing was under bcl2fastq2/2.20.0 and cellranger/2.2.0. We report a increased cDC1 to cDC2 ratio, and an increased inflammatory DC in PAH patient, with the major changes of CXCR4 expression which are experimentally confirmed. This study provide a reference for studying the DC related abonormalties in PAH and other inflammatory diseases.

2022-06-01 | GSE164493 | GEO

Limited systemic sclerosis patients with PAH show biomarkers of inflammation and vascular injury

Project description:Limited systemic sclerosis patients with pulmonary arterial hypertension show biomarkers of inflammation and vascular injury Forty-nine PBMC samples were obtained from 21 lSSc subjects without PAH (lSSc-noPAH), 15 lSSc subjects with PAH (lSSc-PAH), and 10 healthy controls; three subjects provided PBMCs one year later. Genome-wide gene expression was measured for each sample. Gene expression clearly distinguished lSSc samples from healthy controls, and separated lSSc-PAH from lSSc-NoPAH patients. The gene expression and cytokine profiles of lSSc-PAH patients suggest the presence of activated monocytes, and show markers of vascular injury and inflammation. Sample vs reference, total RNA isolated from peripheral blood mononuclear cells (PBMC), 21 lSSc subjects without PAH (lSSc-noPAH), 15 lSSc subjects with PAH (lSSc-PAH), and 10 healthy controls

2010-10-27 | E-GEOD-19617 | biostudies-arrayexpress

Limited systemic sclerosis patients with PAH show biomarkers of inflammation and vascular injury

2010-10-27 | GSE19617 | GEO

Comprehensive transcriptome analysis of the heart in control, PAH and PAH/aKO mice.

Project description:Pregnancy-associated hypertensive (PAH) mice were maintained by mating females carrying the human angiotensinogen (hAGT) gene with males expressing the human renin (hRN) gene, as previously described (Takimoto E., et al., Science, 1996). Angiotensin II (AngII) has critical roles in regulation of blood pressure. In late pregnancy of PAH mice, increased AngII causes acute and severe hypertension with proteinuria. Furthermore, PAH mice show cardiac hypertrophy, fibrosis and apoptosis. It is known that AngII downregulates mRNA of alpha 1a-adrenergic receptor (Adra1a) in neonatal rat cardiac myocytes (Li H.T., et al., Circ. Res., 1997). Interestingly, we found that Adra1a knock out PAH (PAH/aKO) mice display more severe phenotype of cardiac hypertrophy in comparison to PAH mice. In this study, to understand the molecular basis of cardiac hypertrophy via regulation of Adra1a expression with AngII in PAH mice, we performed a comprehensive analysis of gene expression changes in cardiac remodeling of PAH and PAH/aKO mice using the next-generation RNA sequencing (RNA-seq).

2023-01-27 | GSE140948 | GEO

Reversible and irreversible PAH in a rat model for MCT+Shunt-induced PAH and hemodynamic unloading.

Project description:PAH was induced by 60mg/kg MCT and an aorto-caval shunt. At different timepoints of PAH progression (day 14, 21 and 28 after MCT-injection), the left lung with PAH was hemodynamically unloading by unilateral orthotopic transplatation into a syngeneic, healthy recipient. All day 14 and 7/10 day 21 transplanted lungs showed reversal of PAH after LTx. All day 28 and 3/10 day 21 transplanted lungs showed PAH progression after LTx. Lung tissue of Reversible and Irreversible PAH and normal controls, acquired at LTx, was compared using RNA-seq.

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Expression data analysis in lungs from mice induced for pulmonary arterial hypertension (PAH) by inhalation of Stachybotrys chartarum spores

2010-07-29 | GSE23178 | GEO

Expression data analysis in lungs from mice induced for pulmonary arterial hypertension (PAH) by inhalation of Stachybotrys chartarum spores

Project description:It has been reported that repeated intra-tracheal instillation of S. chartarum spores induced significant pulmonary arterial remodeling in mice, which resulted in pathological changes like human pulmonary arterial hypertension (PAH) and elevation right ventricle systolic pressure. Then, we used microarrays to know the complex molecular mechanisms that underlie pathogenesis of PAH. Isolates of Stachybotrys chartarum were used. Ddy mice were anesthetized and the spore suspension was intratracheally injected 12 times, i.e. 1×104 spores into each mouse at 4-5 day intervals for 8 weeks. Mice were sacrificed one week after the final injection and then examined. Lung tissue specimens from mice model of PAH (n=3) and normal controls (n=3) were obtained. RNA targets preparation were performed according to the manufacturer's protocol using GeneChip(R) 3' IVT Express Kit (Affymetrix). One hundred nanograms of total RNA was converted into double-stranded cDNA template for transcription. In vitro transcription synthesized amplified RNA (aRNA) and incorporated a biotin-conjugated nucleotide. After purification and fragmentation of aRNA, 12.5 ug of them was hybridized to GeneChip(R) Mouse Genome 430 2.0 Array (Affymetrix).The Probe Array was scanned using a GeneChip(R) Scanner 3000 7G.

2010-07-28 | E-GEOD-23178 | biostudies-arrayexpress

Polyomic elucidation of the 9,10-anthraquinone biodegradation pathway by a newly isolated oxy-PAH degrading specialist from PAH-contaminated soil

Project description:We report the isolation of the first bacterial strain, Sphingobium sp. AntQ-1, with the ability to grow on the oxy-PAH anthraquinone as sole carbon and energy source. We combine genomic, transcriptomic and metabolomic analysis to comprehensively elucidate the anthraquinone metabolic pathway and to identify key degradative genes.

2023-01-24 | GSE199781 | GEO

miRNA profiling in hPASMC isolated from normal and PAH patients

Project description:To identify dysregulated miRNAs in PAH HPASMC, we compared the miRNA expression profiles between normal and PAH HPASMC using the RT2 miRNA PCR Array System. Total RNA was isolated using a miRNeasy Mini Kit (Qiagen, Valencia, CA) and treated with an RNase-Free DNase Set (Qiagen). After quantification with Nanodrop 2000 spectrophotometer (ThermoScientific, Rockford, IL), miRNAs were reversely transcribed using a RT2 miRNA First Strand Kit (SABiosciences, Frederick, MD). The RT2 miRNA PCR Array System (SABiosciences) was used to study miRNA profiling in normal and PAH HPASMC. U6, SNORD44, SNORD47 and SNORD48 were used as internal controls for the profiling. HPASMC isolated from oe normal donor (C128, as control), one IPAH patient (B157), one PAH patient with Eisenmenger’s syndrome (CC-008) and one PAH patient with Scleroderma (CCF-004) were used for miRNA PCR array study. Data is normalized to the control sample. Three independent sample preparations were used for this study.

2014-02-28 | E-GEOD-55427 | biostudies-arrayexpress

Human Neutrophils in Pulmonary Arterial Hypertension by LC-MSMS

Project description:Neutrophil elastase (NE) is implicated in pulmonary arterial hypertension (PAH) but the role of neutrophils in the pathogenesis of PAH is unclear. Here we show that neutrophils from PAH vs. control subjects produce and release increased NE associated with enhanced extracellular trap formation. PAH neutrophils are highly adherent and show decreased migration consistent with increased vinculin, identified on proteomic analysis and previously linked to an antiviral response. This was substantiated by a transcriptomic interferon signature in PAH neutrophils and an increase in human endogenous retrovirus (HERV-K) envelope protein. NE and interferon genes are induced by HERV-K envelope and vinculin is increased by HERV-K dUTPase that is elevated in PAH plasma. Neutrophil exosomes from PAH plasma contain increased NE and HERV-K envelope and induce pulmonary hypertension in mice, that is mitigated by the NE inhibitor and antiviral agent, elafin. Thus elevated HERVs explain pathological neutrophils linked to PAH induction and progression.

2023-03-11 | PXD023134 | Pride

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