Project description:In atherosclerosis progression and regression, monocytes or monocyte-derived macrophages are the major immune cells in the plaque. It is important to understand the fate and characteristics of monocyte/macrophage during the plaque progression and regression. To characterize the fate of monocytes/macrophages, we performed single cell RNA sequencing of fate-mapped aortic CX3CR1-derived monocytes/macrophages from Cx3cr1CreERT2-IRES-YFP/+Rosa26floxed-tdTomato/+ mice with AAV-PCSK9 injection and fed a Western Diet. The single cell RNA-seq analyses revealed the heterogeneity of aortic macrophages and identified a stem-like cell cluster in atherosclerotic aorta.
Project description:Atherosclerosis is modulated by immune cells. Understanding the diversity and heterogeneity in the aorta in steady state, as well as during different stages of disease will help to determine mechanisms promoting disease progression. To characterize the immune cells, we performed single cell RNA sequencing of aortic CD45+ leukocytes from C57BL/6 (WT) on chow diet and early disease Ldlr-/- mice fed high fat diet (HFD) for 21-days. The single cell RNA-seq analyses revealed the heterogeneity of aortic macrophages in steady state and early atherosclerotic disease.
Project description:Atherosclerosis is a systemic and chronic inflammatory disease propagated by monocytes and macrophages. Yet, our knowledge on how transcriptional profiles of these cells evolve in time and space is limited. We aimed at characterizing gene expression changes in site-specific macrophages, i.e. atherosclerotic aorta versus remote peritoneal cavity, and in circulating monocytes during the course of atherosclerosis initiation and progression in Apoe deficient mice. Constructing a comparative directory of cell type- and disease stage-specific whole transcriptome information, we identified a distinctive gene regulation profile of aortic macrophages. Overall 1149 differentially expressed genes (DEG) were involved in the biological modulation of aortic macrophages, and they were further categorized into regulated early, late, and transitorily throughout the disease course. The commonality of gene regulation was surprisingly low among the three investigated cell types. Through complementary interrogation of murine and human single cell RNA sequencing datasets, we showcased the practicality of our directory, using the selected gene, Glycoprotein Nmb (Gpnmb), whose expression in aortic macrophages, and a subset of foamy macrophages in particular, strongly correlated with disease advancement during atherosclerosis initiation and progression. Our study provides a unique toolset to explore gene regulation of macrophage-related biological processes in and outside the atheromatous plaque at early and advanced disease stages for pathomechanistic insight and therapeutic target screening.
2023-04-06 | GSE213189 | GEO
Project description:Single cell RNA sequencing of aortic CD45+ cells reveals aortic macrophage heterogeneity in mouse atherosclerosis
Project description:In atherosclerosis, several immune cells are involved in plaque formation. Foam cell formation is a major cellular process in atherosclerotic lesion. It is important to understand which cells participate in foam cell formation. To characterize the immune cells and foam cells in atherosclerotic aorta, we performed single cell RNA sequencing of aortic CD45+ leukocytes from Ldlr-/- mice and foamy cells from ApoE-/- mice. The single cell RNA-seq analyses revealed the heterogeneity of aortic macrophages and foam cells in atherosclerotic aorta.
Project description:We have applied single-cell RNA sequencing as an unbiased profiling strategy to interrogate and classify aortic macrophage heterogeneity at the single-cell level in atherosclerosis.
Project description:Objective: Disturbed blood flow characterized by low and oscillatory shear stress and complex microenvironment is one of the causes of the atherosclerosis progression. During the development, endothelial cells generally develop into a dysfunction state, so do other cells. To further analyze the heterogeneity of different cell types, single-cell RNA sequencing was used to provide detailed transcriptomic information.</p><p>Approach and results: Transcriptional analysis of single cells from aortic arch of 16-week Apoe-/- mice under normal and atherosclerotic condition was carried out using single-cell RNA sequencing. By unsupervised clustering, 10 types of cells were identified: endothelial cells, immune cells, fibroblasts, erythroids, smooth muscle cells, cycling basal cells, clara cells, mesothelial cells, adipocytes and neurons. Among these cells, endothelial cells and immune cells exhibited great heterogeneity especially between normal and atherosclerotic group. Gene expression profile of later screened cells identified ten subpopulations of endothelial cells of different functions. Distribution of these cells in the control group and HFD group varies. Immune cells consist of macrophages, foam cells, B cells, T cells, neutrophils and NK cells mainly. During the data processing, the cell sorting is done by recognition with specific markers and rules.</p><p>Conclusions: Our data provide a comprehensive description of all kinds of cells in aortic arch and the heterogeneity brought by western diet. These findings open a new sight into the transcriptional development of atherosclerosis.
