MicroRNA profiling of cells isolated from human abdominal aortic aneurysms. Let-7f, a new potential circulating biomarker
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ABSTRACT: This study aims to identify and characterize miRNA expression in aneurysmal smooth muscle cells (SMCs) and M1 and M2 macrophages isolated by laser microdissection from human AAA biopsy samples. The aim of this study was to profile (with microarray technology) miRNAs in M1 and M2 macrophages and Smooth muscle cells (SMCs), isolated by laser capture microdissection (LCM). SMCs isolated from control non-aneurysmal aortas were the control group according to which data were normalized. Areas enriched in M1 macrophages, M2 macrophages and smooth muscle cells were microdissected (9.8 [4.7-16.2] mm2) from two different biopsy samples of human abdominal aortic aneurysm. An area enriched in smooth muscle cells was microdissected from two biopsy samples of non aneurysmal abdominal aortas. Each microdissected samples were analyzed independently on microarray. M1 and M2 macrophages and smooth muscle cells expressed in human abdominal aneurysmal aortas were each analyzed in duplicate (each isolated from different human donors of abdominal aortic aneurysm). Analysis of replicated samples of cells were performed on a second microarray. Data were normalized with smooth muscle cells isolated from human abdominal non aneurysmal aortas.
Project description:This study aims to identify and characterize miRNA expression in aneurysmal smooth muscle cells (SMCs) and M1 and M2 macrophages isolated by laser microdissection from human AAA biopsy samples. The aim of this study was to profile (with microarray technology) miRNAs in M1 and M2 macrophages and Smooth muscle cells (SMCs), isolated by laser capture microdissection (LCM). SMCs isolated from control non-aneurysmal aortas were the control group according to which data were normalized. Areas enriched in M1 macrophages, M2 macrophages and smooth muscle cells were microdissected (9.8 [4.7-16.2] mm2) from two different biopsy samples of human abdominal aortic aneurysm. An area enriched in smooth muscle cells was microdissected from two biopsy samples of non aneurysmal abdominal aortas. Each microdissected samples were analyzed independently on microarray.
Project description:This study aims to identify and characterize miRNA expression inATLOs isolated by laser microdissection from human AAA biopsy samples. The aim of this study was to profile (with microarray technology) miRNAs in ATLOs (Adventitial tertiary lymphoid organs), isolated by laser capture microdissection (LCM). Smooth muscle cells (SMCs) isolated from control non-aneurysmal aortas were the control group according to which data were normalized. ATLOs were microdissected (mean 13.5mm2) from two different biopsy samples of human abdominal aortic aneurysm. An area enriched in smooth muscle cells was microdissected from two biopsy samples of non aneurysmal abdominal aortas. Each microdissected samples were analyzed independently on microarray. ATLOs located in human abdominal aneurysmal aortas were each analyzed in duplicate (each isolated from different human donors of abdominal aortic aneurysm). Data were normalized with smooth muscle cells isolated from human abdominal non aneurysmal aortas.
Project description:This study aims to identify and characterize miRNA expression inATLOs isolated by laser microdissection from human AAA biopsy samples. The aim of this study was to profile (with microarray technology) miRNAs in ATLOs (Adventitial tertiary lymphoid organs), isolated by laser capture microdissection (LCM). Smooth muscle cells (SMCs) isolated from control non-aneurysmal aortas were the control group according to which data were normalized. ATLOs were microdissected (mean 13.5mm2) from two different biopsy samples of human abdominal aortic aneurysm. An area enriched in smooth muscle cells was microdissected from two biopsy samples of non aneurysmal abdominal aortas. Each microdissected samples were analyzed independently on microarray.
Project description:Inflammation and elastin degradation are key hallmarks in the pathogenesis of abdominal aortic aneurysms (AAA). It has been acknowledged that activation of alpha7 nicotinic acetylcholine receptors (α7nAChRs) attenuates inflammation, termed the cholinergic anti-inflammatory pathway (CAP). Thus, we hypothesized that low-dose nicotine, an α7nAChR agonist, impairs the progression of AAAs in rats by activating the CAP. Male Sprague-Dawley rats underwent surgical AAA induction with intraluminal elastase infusion. We compared vehicle rats with rats treated with nicotine (1.25 mg/kg/day) and the aneurysm progression was monitored by weekly ultrasound images for 28 days. Nicotine significantly promoted AAA progression after 28 days of treatment (p=0.031). Additionally, gelatin zymography demonstrated that nicotine significantly reduced pro-matrix metalloprotease (pro-MMP) 2(p=0.029) and MMP9(p=0.030) activity in aneurysmal tissue. No significant difference was found in elastin content or the score of elastin degradation between the groups. Neither infiltrating neutrophils nor macrophages, nor aneurysmal messenger RNA (mRNA) levels of pro- or anti-inflammatory cytokines differed between the vehicle and nicotine group. Finally, no difference in mRNA levels of markers for antioxidative stress or vascular smooth muscle cells contractile phenotype was observed. However, proteomics analyses of non-aneurysmal abdominal aortas revealed that nicotine decreased proteins in the ontology terms inflammation and reactive oxygen species and in contradiction to augmented AAAs. In conclusion, treatment with the given nicotine dose augmented AAA progression in this rat model.
Project description:We report bulk RNA-seq data of Myh11 lineage marked smooth muscle cells (SMCs) from murine aortas at 1 week after conditional disruption of TGFβ signaling versus controls
Project description:We report single-cell RNA-seq data of Wnt1-Cre2 lineage marked, integrin alpha 8 labeled smooth muscle cells (SMCs) from murine aortas at 12 weeks of age
Project description:We report single-cell RNA-seq data of Nkx2-5 lineage marked, integrin alpha 8 labeled smooth muscle cells (SMCs) from murine aortas at 12 weeks of age
Project description:We report single-cell RNA-seq data of Myh11 lineage marked smooth muscle cells (SMCs) from murine aortas at 1 week after conditional disruption of TGFβ signaling versus controls
Project description:MiR-30e represses the osteogenic program in mesenchymal stem cells (MSCs) and smooth muscle cells (SMCs) by targeting IGF2, and drives their differentiation into adipogenic or smooth muscle lineage, respectively. In Experiment 1, SMCs were cultured from mouse aortas and transduced with miR-30e or control lentivirus. In Experiment 2, MSCs were extracted from mouse bone marrow and transduced with miR-30e or control lentivirus. In Experiment 3, the MSCs were transfected with a scrambled oligo or anti-miR-30e oligo. In all 3 experiments, N=3 per group. RNA was extracted from each experiment and run on Affymetrix arrays.
Project description:MiR-30e represses the osteogenic program in mesenchymal stem cells (MSCs) and smooth muscle cells (SMCs) by targeting IGF2, and drives their differentiation into adipogenic or smooth muscle lineage, respectively. In Experiment 1, SMCs were cultured from mouse aortas and transduced with miR-30e or control lentivirus. In Experiment 2, MSCs were extracted from mouse bone marrow and transduced with miR-30e or control lentivirus. In Experiment 3, the MSCs were transfected with a scrambled oligo or anti-miR-30e oligo. In all 3 experiments, N=3 per group. RNA was extracted from each experiment and run on Affymetrix arrays.