ABSTRACT: The Total RNA from seven control pool and seven atherosclerotic plaque sample pools were labeled by Cy3/Cy5 respectively and analyzed on 22K human genome array chip V1.0.
Project description:To investigate the expression profiles of miRNA in atherosclerotic plaques, the global features of miRNAs expression of three normal coronary artery tissues sample pools and three sample pools of advanced atherosclerosis plaques of coronary artery were studied using microarray technology,
Project description:The goal of the project is the detection of region-specific lipid features that discriminate between symptomatic and asymptomatic human carotid atherosclerotic plaques by MALDI MSI.
Project description:Cardiovascular disease (CVD) is the leading cause of death, with atherosclerosis a major underlying cause. While often asymptomatic for decades, plaque destabilisation and rupture can arise suddenly and cause acute arterial occlusion or peripheral embolisation resulting in acute myocardial infarction, stroke and lower limb ischaemia. Hard plaques are typically considered as stable, and soft plaques as unstable. Extracellular matrix (ECM) remodelling can result in plaque destabilisation, but the mechanisms that drive the development of unstable lipid-rich plaques with a thin fibrous cap, versus stable fibrotic plaques with a thick cap, are not fully understood. We hypothesised that there would be significant differences in ECM composition between hard (stable) and soft (unstable and rupture-prone) plaques. We identified and quantified >46700 proteins, including 367 ECM proteins, with unprecedented coverage and high reproducibility. We identified 575 proteins with differential abundances between hard (stable) and soft (unstable) plaques. Proteins involved in inflammation and ECM remodeling, including multiple proteases were enriched, and ECM proteins decreased, in soft plaques. These data provide a unique insight into inflammatory mechanisms and ECM remodelling as an explanation for plaque destabilization. Furthermore they provide a first step towards identifying circulating biomarkers for individualised risk profiling of arteriosclerosis.
Project description:Abnormal epigenetic gene regulation may play a pivotal role in atherogenesis. In particular, global DNA hypomethylation potentially leading to proatherogenic gene expression, occurs in atherosclerotic lesions in humans and animal models. In order to identify genomic sequences targeted for DNA hypomethylation in atherosclerosis, we analysed the methylation status of CpG islands (CGIs) in 45 human arteries with advanced atherosclerotic lesions and 16 normal counterparts by a microarray approach. Methylation data for 10,367 CGIs revealed that a subset (1.5%) of such sequences was hypermethylated in normal arteries, in accordance with data previously obtained in peripheral blood cells. Ninety-four per cent of this CGI subset was demethylated in atherosclerotic tissue, while only 17 of normally hypomethylated CGIs was hypermethylated in diseased tissue. A functional classification of genes physically associated with differentially methylated CGIs revealed a bias towards transcription factors (42%). The latter include HOX members, NOTCH1 and FOXP1, which are known to regulate angiogenesis, dedifferentiation, cell migration and macrophage function. The methylation status of selected CGIs was validated in further 10 subjects for either group. Expression patterns of these factors were compatible with the observed differential methylation. Our data suggest that one of the molecular changes associated with aberrant DNA methylation in advanced atherosclerosis is the regulation of critical transcription factor genes for the induction of a proatherogenic cellular phenotype. Two-condition experiment, i.e. normal vs. atherosclerotic arteries. 16 and 45 normal and atherosclerotic samples, respectively, were pooled and used to interrogate CpG island arrays in triplicate, for a total of six arrays. Each array was co-hybridized with untreated DNA (reference) and hypermethylated DNA obtained by biochemical filtration.
Project description:We compared gene expression profiles between asymptomatic and symptomatic atherosclerotic plaques from the same patient. This was accomplished by analyzing carotid plaques from four patients with bilateral high-grade carotid artery stenoses one being symptomatic (TIA or stroke) and the other asymptomatic.
Project description:Although it is well known that the extracellular matrix composition significantly impacts the development of neointimal hyperplasia and atherosclerosis, the biochemical cues regulating the cell-ECM interactions within the neointima remain obscure. Here, we pairwise collected 14 atherosclerotic plaques excised during the carotid endarterectomy and 14 adjacent intact aortic segments and then interrogated their proteomic profile.
Project description:Bulk RNAsequencing of CD4+ T-cells in SLE patients with (SLE-P) and without (SLE-NP) subclinical atherosclerotic plaques in carotid and femoral arteries, scanned by vascular ultrasound.
Project description:Bulk RNAsequencing of CD8+ T-cells in SLE patients with (SLE-P) and without (SLE-NP) subclinical atherosclerotic plaques in carotid and femoral arteries, scanned by vascular ultrasound.
Project description:Recent findings have challenged the prevailing histology- or imaging-based definition of the “vulnerable plaque”. To investigate molecular characteristics associated with “clinical instability” of atherosclerosis, we performed a proteomics comparison of the vascular extracellular matrix and associated molecules in human carotid endarterectomy specimens from symptomatic versus asymptomatic patients. The proteomics data were integrated with gene expression profiling and an analysis of protein secretion by lipid-loaded human vascular smooth muscle cells. The molecular signature of plaques from symptomatic patients identified by proteomics and at least one of the other two approaches comprised matrix metalloproteinase-9, chitinase-3-like protein 1, S100A8/S100A9, cathepsin B, fibronectin and galectin-3-binding protein. These biomarker candidates were measured in 685 subjects of the Bruneck Study and found to significantly predict the progression to advanced atherosclerosis (as assessed by repeated carotid ultrasound) and the incidence of cardiovascular disease over a 10-year follow-up period, highlighting the strength of tissue-based proteomics for biomarker discovery.
Project description:Matrix metalloproteinase 12 (MMP12) is a macrophage-secreted protein that is massively upregulated as a pro-inflammatory factor in metabolic and vascular tissues of mice and humans suffering from cardiometabolic diseases (CMDs). However, the molecular mechanisms explaining the contributions of MMP12 to CMDs are still unclear.