S-EPMC6594632 - Isolation, culturing and gene expression profiling of inner mass cells from stable and vulnerable carotid atherosclerotic plaques. - OmicsDI

Unknown

Dataset Information

Isolation, culturing and gene expression profiling of inner mass cells from stable and vulnerable carotid atherosclerotic plaques.

ABSTRACT: The connective tissue components that form the atherosclerotic plaque body are produced by the plaque inner mass cells (PIMC), located inside the plaque. We report an approach to isolate and culture cells from the connective tissue of stable and vulnerable human atherosclerotic plaques based on elimination of non-connective tissue cells such as blood and non-plaque intima cells with a lysis buffer. The resulting plaque cells were characterized by growth capacity, morphology, transcriptome profiling and specific protein expression. Plaque cells slowly proliferated for up to three passages unaffected by the use of proliferation stimulants or changes of culture media composition. Stable plaques yielded more cells than vulnerable ones. Plaque cell cultures also contained several morphological cellular types. RNA-seq profiles of plaque cells were different from any of the cell types known to be involved in atherogenesis. The expression of the following proteins was observed in cultured plaque cells: smooth muscle cells marker α-SMA, macrophage marker CD14, extracellular matrix proteins aggrecan, fibronectin, neovascularisation markers VEGF-A, CD105, cellular adhesion receptor CD31 and progenitor/dedifferentiation receptor CD34. Differential expression of several notable transcripts in cells from stable and vulnerable plaques suggests the value of plaque cell culture studies for the search of plaque vulnerability markers.

SUBMITTER: Novikova OA

PROVIDER: S-EPMC6594632 | biostudies-literature | 2019

REPOSITORIES: biostudies-literature

altmetric image

Publications

Isolation, culturing and gene expression profiling of inner mass cells from stable and vulnerable carotid atherosclerotic plaques.

Novikova Olga A OA Nazarkina Zhanna K ZK Cherepanova Anna V AV Laktionov Petr P PP Chelobanov Boris P BP Murashov Ivan S IS Deev Roman V RV Pokushalov Evgeny A EA Karpenko Andrey A AA Laktionov Pavel P PP

PloS one 20190626 6

The connective tissue components that form the atherosclerotic plaque body are produced by the plaque inner mass cells (PIMC), located inside the plaque. We report an approach to isolate and culture cells from the connective tissue of stable and vulnerable human atherosclerotic plaques based on elimination of non-connective tissue cells such as blood and non-plaque intima cells with a lysis buffer. The resulting plaque cells were characterized by growth capacity, morphology, transcriptome profil ...[more]

Similar Datasets

Project description:Atherosclerosis is a chronic inflammatory disease with high prevalence and mortality. The rupture of atherosclerotic plaque is the main reason for the clinical events caused by atherosclerosis. Making clear the transcriptomic and proteomic profiles between the stabe and unstable atherosclerotic plaques is crucial to prevent the clinical manifestations. In the present study, 5 stable and 5 unstable human carotid atherosclerotic plaques were obtained by carotid endarterectomy. The samples were used for the whole transcriptome sequencing (RNA-Seq) by the Next-Generation Sequencing using the Illumina HiSeq, and for proteome analysis by HPLC-MS/MS. The lncRNA-targeted genes and circRNA-originated genes were identified by analyzing their location and sequence. Gene Ontology and KEGG enrichment was carried out to analyze the functions of differentially expressed RNAs and proteins. The protein-protein interactions (PPI) network was constructed by the online tool STRING. The consistency of transcriptome and proteome were analyzed, and the lncRNA/circRNA-miRNA-mRNA interactions were predicted. As a result, 202 mRNAs, 488 lncRNAs, 91 circRNAs, and 293 proteins were identified to be differentially expressed between stable and unstable atherosclerotic plaques. The 488 lncRNAs might target 381 protein-coding genes by cis-acting mechanisms. Sequence analysis indicated the 91 differentially expressed circRNAs were originated from 97 protein-coding genes. These differentially expressed RNAs and proteins were mainly enriched in the terms of the cellular response to stress or stimulus, the regulation of gene transcription, the immune response, the nervous system functions, the hematologic activities, and the endocrine system. These results were consistent with the previous reported data in the dataset GSE41571. Further analysis identified CD5L, S100A12, CKB (target gene of lncRNA MSTRG.11455.17), CEMIP (target gene of lncRNA MSTRG.12845), and SH3GLB1 (originated gene of hsacirc_000411) to be critical genes in regulating the stability of atherosclerotic plaques. Our results provided a comprehensive transcriptomic and proteomic knowledge on the stability of atherosclerotic plaques.

Project description:To compare the characteristics of stable and vulnerable carotid plaques, and investigate the diagnostic performance of wall shear stress (WSS) based on magnetic resonance plaque imaging in carotid plaques. Retrospectively analyzed and divided 64 atherosclerotic plaques into stable carotid plaque groups with mild-to-moderate stenosis and vulnerable carotid plaque groups with significant stenosis. Computational fluid dynamics simulations were performed to calculate WSS parameters by using three-dimensional wall geometry based on high-resolution magnetic resonance plaque imaging of carotid bifurcation and patient specific boundary conditions obtained through color Doppler ultrasound. WSS parameters including upstream (WSSup), downstream (WSSdown), and core (WSScore) of plaque. The WSS parameters values were compared between the stable and vulnerable carotid plaque groups. Receiver operating characteristic curves and area under the curve (ROC-AUC) and Python were used to evaluate discriminative efficacy of WSS. WSSdown exhibited significant decrease in the vulnerable carotid plaque group (2.88 ± 0.41 Pa) compared to the stable carotid plaque group (4.47 ± 0.84 Pa) (P = .003). The difference of WSSup (3.28 ± 0.85 Pa vs 4.02 ± 0.74 Pa) and WSScore (1.12 ± 0.18 Pa vs 1.38 ± 0.38 Pa) between the two groups were also pronounced (P = .02, 0.01, respectively). The ROC-AUC values for WSSup, WSSdown, WSScore were 0.75 (95% CI, 0.58-0.93), 0.96 (95% CI, 0.79-1.14), 0.69 (95% CI, 0.56-0.83) respectively. When the value of WSSdown was 3.5 Pa, the sensitivity was 93.7% (95% CI, 76.1-111), specificity and accuracy was 87.5% (95% CI, 70.0-105), 88.4% (95% CI, 70.6-105) respectively. Notably, among these parameters, WSSdown demonstrated the highest discriminative efficiency with a F1 Score of 0.90, Diagnostic Odds Ratio of 105.0 and Matthews Correlation Coefficient of 0.81. Vulnerable carotid plaques with significant stenosis have lower WSS compared to stable plaques with mild-to-moderate stenosis, and downstream WSS showing the highest diagnostic efficacy.

Project description:AimsCardiovascular disease remains a major global health concern, with atherosclerosis (AS) being a significant contributor. Vulnerable plaques play a critical role in acute cardiovascular events. Syndecan-1 (SDC-1), a vital membrane proteoglycan in the vascular endothelial glycocalyx, is believed to be associated with plaque progression. However, its precise relationship with severity and vulnerability of atherosclerotic plaque remains unclear. This study aimed to investigate SDC-1 expression and its potential correlation with plaque vulnerability in ApoE-/- atherosclerosis mouse model.Methods and resultsEight-week-old mice were induced into the AS model using a high-fat diet (HFD) and/or partial ligation of the left common carotid artery (PLCA), with a chow diet (CD) control group. After 16 weeks, plaques in the aortic root showed the following order: HFD + PLCA group > HFD group > CD + PLCA group > CD group. Immunohistochemistry revealed heightened accumulation of lipid/foam cells and CD68-labeled macrophages in the plaques, elevated vascular endothelial growth factor (VEGF), and matrix Metalloproteinase-9 (MMP-9) in the HFD + PLCA group's plaques, along with reduced collagen and α-SMA-labeled smooth muscle cells, resulting in the highest vulnerability index value. Immunohistofluorescence analysis of frozen plaque sections showed significantly higher SDC-1 expression in the AS mice group compared to the CD group, both positively correlated with plaque vulnerability. Serum analysis demonstrated elevated levels of SDC1, sphingosine 1-phosphate (S1P), and VEGF-A in the AS mice, all positively correlated with plaque vulnerability. Multivariate analysis identified SDC1 as an independent predictor of plaque vulnerability.ConclusionThis study enhances our understanding of plaque vulnerability mechanisms and presents SDC1 as a potential biomarker for atherosclerosis. These findings underscore the importance of addressing modifiable risk factors, such as diet and hemodynamics and suggest the utility of serum SDC1 as a valuable clinical marker. Ultimately, these insights may lead to more effective strategies in combating cardiovascular diseases and improving patient outcomes.