Single-cell RNA sequencing of bone marrow HSPCs isolated from patients after myocardial infarction and control patients
Ontology highlight
ABSTRACT: This experiment focused on studying the impact of myocardial infarction on bone marrow HSPCs (LineageNeg, CD38Neg, CD34Pos). Human sternal bone marrow was extracted from the sternum of acute patients who had undergone myocardial infarction (n = 6), as well as from control patients (n = 7), during surgical procedures.
Project description:This experiment focused on studying the impact of myocardial infarction on bone marrow monocytes. Human sternal bone marrow was extracted from the sternum of acute and chronic patients who had undergone myocardial infarction, as well as from control patients, during surgical procedures. The samples underwent Ficoll separation and were subsequently frozen for preservation. On the day of sorting, the samples were thawn, and a total of 3,000 - 20,000 human bone marrow monocytes (Lineage-, CD45+, HLA-DR+) were sorted in 2% BSA in PBS in two biological replicates. scRNA-seq was performed.
Project description:Study of the transcriptomic changes in human HSPCs under the influence of at-RA and 4-oxo-RA. Human Hematopoietic Stem and Progenitor Cells (HSPCs)(Lineage negative, CD38 negative, CD34 positive cells) were isolated from sternal bone marrow of patients who had experienced myocardial infarction or from control patients. For in vitro cultivation, cells were treated with at-RA (2.5 μM final concentration), 4-oxo-RA (2.5 μM final concentration), or the respective volume of DMSO (control) for 72 hours. After incubation period, cells were harvested for RNA extraction.
Project description:Study of the impact of 4-oxo-RA on LSK (bone marrow and spleen) cell characteristics post-myocardial infarction in C57BL/6J mice. Myocardial infarction was induced in female C57BL/6J mice (aged 6 to approximately 12 weeks) through LAD occlusion. Post-surgery, mice were intraperitoneally injected with 30 mg/kg 4-oxo-RA (MI+4-oxo-RA) or the equivalent DMSO in PBS (MI+vehicle) on the 1st and 2nd days after MI. Three days post-MI, mice were euthanized. ScRNA-seq was performed on 25k LSK cells (Lin-, Sca1+, c-Kit+) that were facs sorted.
Project description:Study of the impact of 4-oxo-RA on LSK cell characteristics post-myocardial infarction in RarB KO mice. Myocardial infarction was induced in female RarB KO mice aged 6 to 12 weeks by LAD occlusion. On the 1st and 2nd post-MI days, mice were intraperitoneally injected with 30 mg/kg 4-oxo-RA (MI+4-oxo-RA) or DMSO in PBS (MI+vehicle). Three days post-MI, mice were euthanized. ScRNA-seq was performed on 25k LSK cells (Lin-, Sca1+, c-Kit+) that were facs sorted.
Project description:Bone marrow and peripheral blood were obtained from patients after myocardial infarction. Erythrocytes were removed by NH4CL lysis to finally obtain nucleated bone marrow cells and peripheral blood leukocytes. After RNA isolation, RNA from three patients was pooled and the RNA expression profile of both organism parts were compared.
Project description:We demonstrate an age-independent loss of type H bone endothelium in heart failure after myocardial infarction in both mice and in humans. Using single-cell RNA sequencing, we delineate the transcriptional heterogeneity of human bone marrow endothelium showing increased expression of inflammatory genes, including IL1B and MYC, in ischemic heart failure. Endothelial-specific overexpression of MYC was sufficient to induce type H bone endothelial cells, whereas inhibition of NLRP3-dependent IL-1 production partially prevents the post-myocardial infarction loss of type H vasculature in mice.
Project description:We demonstrate an age-independent loss of type H bone endothelium in heart failure after myocardial infarction in both mice and in humans. Using single-cell RNA sequencing, we delineate the transcriptional heterogeneity of human bone marrow endothelium showing increased expression of inflammatory genes, including IL1B and MYC, in ischemic heart failure. Endothelial-specific overexpression of MYC was sufficient to induce type H bone endothelial cells, whereas inhibition of NLRP3-dependent IL-1 production partially prevents the post-myocardial infarction loss of type H vasculature in mice.
Project description:Despite a substantial progress in diagnosis and therapy, acute myocardial infarction (MI) is a major cause of mortality in the general population. A novel insight into the pathophysiology of myocardial infarction obtained by studying gene expression should help to discover novel biomarkers of MI and to suggest novel strategies of therapy. The aim of our study was to establish gene expression patterns in leukocytes from acute myocardial infarction patients. ST-segment elevation myocardial infarction alters expression of several groups of genes. On admission, several genes and pathways that could be directly or indirectly linked with lipid/glucose metabolism, platelet function and atherosclerotic plaque stability were affected (signaling of PPAR, IL-10, IL-6). Analysis at discharge highlighted specific immune response (upregulation of immunoglobulins). Highly significant and substantial upregulation of SOCS3 and FAM20 genes expression in the first 4-6 days of myocardial infarction in all patients is the most robust observation of our work Twenty-eight patients with ST-segment elevation myocardial infarction (STEMI) were included. The blood was collected on the 1st day of myocardial infarction, after 4-6 days, and after 6 months. Control group comprised 14 patients with stable coronary artery disease (CAD), without history of myocardial infarction. Gene expression analysis was performed with Affymetrix GeneChipM-BM-. Human Gene 1.0 ST microarrays and GCS3000 TG system.
Project description:Hematopoiesis advances cardiovascular disease by generating inflammatory leukocytes that attack the arteries, heart and brain. While it is well documented that the bone marrow niche regulates hematopoietic stem cell proliferation and hence the systemic leukocyte pool, it is less clear how cardiovascular disease affects the vasculature forming this niche. Here we show that arterial hypertension, atherosclerosis and myocardial infarction alter the anatomy and function of bone marrow vasculature. Hypertension and atherosclerosis instigated vascular fibrosis, leakage and endothelial dysfunction in the bone marrow. Myocardial infarction induced vascular leakage and bone marrow angiogenesis via Vegf signaling. Endothelial cell-specific deletion of the Vegf receptor 2 limited emergency hematopoiesis after myocardial infarction, indicating that new vasculature supports higher blood cell production. RNA-sequencing of bone marrow endothelial cells revealed inflammatory gene expression in mice with cardiovascular disease. Endothelial cell-specific deletion of interleukin 6 or versican, which were highly expressed in mice with atherosclerosis or myocardial infarction, respectively, reduced hematopoiesis and systemic myeloid cells. Taken together, cardiovascular disease affects the vascular bone marrow niche, thus influencing hematopoietic stem cell behavior and expanding innate immune cell supply to atherosclerotic plaque and ischemic myocardium. Interrupting this feed back loop may constrain cardiovascular inflammation.
Project description:Hematopoiesis advances cardiovascular disease by generating inflammatory leukocytes that attack the arteries, heart and brain. While it is well documented that the bone marrow niche regulates hematopoietic stem cell proliferation and hence the systemic leukocyte pool, it is less clear how cardiovascular disease affects the vasculature forming this niche. Here we show that arterial hypertension, atherosclerosis and myocardial infarction alter the anatomy and function of bone marrow vasculature. Hypertension and atherosclerosis instigated vascular fibrosis, leakage and endothelial dysfunction in the bone marrow. Myocardial infarction induced vascular leakage and bone marrow angiogenesis via Vegf signaling. Endothelial cell-specific deletion of the Vegf receptor 2 limited emergency hematopoiesis after myocardial infarction, indicating that new vasculature supports higher blood cell production. RNA-sequencing of bone marrow endothelial cells revealed inflammatory gene expression in mice with cardiovascular disease. Endothelial cell-specific deletion of interleukin 6 or versican, which were highly expressed in mice with atherosclerosis or myocardial infarction, respectively, reduced hematopoiesis and systemic myeloid cells. Taken together, cardiovascular disease affects the vascular bone marrow niche, thus influencing hematopoietic stem cell behavior and expanding innate immune cell supply to atherosclerotic plaque and ischemic myocardium. Interrupting this feed back loop may constrain cardiovascular inflammation.