Effects of Post-Myocardial Infarction Heart Failure on the Bone Vascular Niche (Mouse Data)
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ABSTRACT: 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:Clonal hematopoiesis of indeterminate potential (CHIP) is caused by somatic mutations in hematopoietic stem cells and is associated with worse prognosis in heart failure patients. Patients harboring CHIP mutations show enhanced inflammation. However, whether these signatures are derived from the relatively low number of cells harboring mutations or are indicators of a systemic pro-inflammatory activation that is associated with CHIP is unclear. In this study, we assessed the cell-intrinsic effects of CHIP mutant-carrying cells in patients with heart failure. Using an improved protocol to detect mutant cells on a single cell level (MutDetect-Seq), we show that DNMT3A mutant monocytes exhibit altered gene expression profiles associated with inflammation and phagolysosome function. Gene expression was also significantly altered in DNMT3A mutant CD4+ T cells and NK cells, but not CD8 T cells. DNMT3A silenced CD4+ T cells and NK cells showed increased effector functions. Moreover, increased paracrine signaling pathways are predicted and validated between mutated and wild monocytes and T cells, which amplify inflammatory circuits. Taken together, these data provide novel insights into how CHIP might promote worse prognosis in heart failure patients.
Project description:Cardiac fibrosis is a common feature of ischemic heart disease and cardiac fibroblasts (CF) are key players in cardiac remodeling of the injured heart after myocardial infarction (MI). Fibrosis increases myocardial stiffness, thereby impairing cardiac function, which ultimately progresses to end-stage heart failure. Little is known, however, on the secretome of CF and cell-to-cell communication of CF is only incompletely understood. Here, we in vivo labeled secreted proteins by expressing TurboID under control of the POSTN promotor in cardiac fibroblasts of mouse with myocardial infarction, enriched biotinylated proteins and analyzed them using LC-MS.
Project description:Hematopoietic mutations in epigenetic regulators like DNA methyltransferase 3 alpha (DNMT3A) drive clonal hematopoiesis of indeterminate potential (CHIP) and are associated with adverse prognosis in patients with heart failure (HF). The interactions between CHIP-mutated cells and other cardiac cell types remain unknown. Here, we identify fibroblasts as potential interaction partners of CHIP-mutated monocytes using combined transcriptomic data from peripheral blood mononuclear cells of HF patients with and without CHIP and cardiac tissue. We demonstrate that DNMT3A inactivation augments macrophage-to-cardiac fibroblasts interactions and induces cardiac fibrosis in mice and humans. Mechanistically, DNMT3A inactivation increases the release of heparin-binding epidermal growth factor (EGF)-like growth factor (HB-EGF) to activate cardiac fibroblasts. These findings not only identify a novel pathway of DNMT3A CHIP-driver mutation-induced instigation and progression of HF, but may also provide a rationale for the development of new anti-fibrotic strategies.
Project description:The study aimed to investigate the mechanisms of Jianxin (JX) granules, a Traditional Chinese Medicine formulation, in the treatment of heart failure (HF) using metabolomic profiling. The HF model was established by ligation of left coronary artery. The successfully modeled rats were randomly divided into three groups: the model group, the JX granules group, and Sacubitril/Valsartan (S/V) group. Four weeks after treatment, LV tissue samples were collected. Proteomics was used to identify the differentially expressed proteins and potential pathways. The differential expressed proteins (DEPs) were obtained with Tandem Mass Tags approach. As compared to the control group, 150 DEPs were identified in model rats, comprising 112 up-regulated and 38 down-regulated proteins. Notably, in the JX granules group, 25 DEPs were identified compared to the model rats, encompassing 18 up-regulated and 7 down-regulated proteins. In comparison with the control group, 228 DEPs in the JX granules group were obtained, which comprised of 128 up-regulated and 100 down-regulated proteins. As compared with the control group, BP in the model group was mainly enriched in humoral immune response, positive regulation of apoptotic cell clearance, and the response to metal ions; Meanwhile, CC was mainly related to contractile and stress fibers, actin filaments, and extracellular space; MF was mainly enriched in the serine-type peptidase activity, actin binding, and peptidase activity. Furthermore, DEPs indicated by KEGG analysis were primarily linked to pathways such as the complement and coagulation cascades, regulation of the actin cytoskeleton, and adrenergic signaling. In contrast to the model group, BP in JX granules group was primarily associated with processes involving nucleosome organization, chromatin assembly or disassembly, DNA conformation change and small molecule catabolic processes; CC was primarily related to the protein-DNA complex, mitochondrion, intracellular, nucleosome, cytoplasm; MF displayed significant enrichment in protein dimerization and homodimerization activity, catalase activity, chromatin and nucleosomal DNA binding. Considering KEGG enrichment information, DEPs in the JX granules group were mainly enriched in metabolic pathways and ribosome pathway. The key differential genes, such as triosephosphate isomerase 1 (TPI1), lactate dehydrogenase B (LDHB), pyruvate kinase M (PKM), protein kinase B (Akt), and lactate dehydrogenase A (LDHA) were identified. The vital pathways including carbon metabolism, the PI3K-Akt signaling pathway, pyruvate metabolism, and HIF-1α signaling pathway were obtained.
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:Angiotensin-(1-7) (Ang-(1-7)) is an endogenous heptapeptide from the renin-angiotensin system. The cardioprotective role of Ang-(1-7) has been described due to its anti-inflammatory and anti-fibrotic activities. In this context, we investigated the impact of the oral formulation of Ang-(1-7) vehiculized in hydroxypropyl β-cyclodextrin (HPβCD) on cardiac proteome remodeling after experimental myocardial infarction. For this, Wistar male rats were submitted to short- (7 days) or long-term (60 days) oral treatment with HPβCD/Ang-(1-7) after induction of experimental myocardial infarction (MI)
Project description:Although bone marrow-derived mononuclear cells (BMNC) have been extensively used in cell therapy for cardiac diseases, little mechanistic information is available to support reports of their efficacy. To address this shortcoming, we compared structural and functional recovery and associated global gene expression profiles in post-ischaemic myocardium treated with BMNC transplantation. BMNC suspensions were injected into cardiac scar tissue 10 days after experimental myocardial infarction. Six weeks later, mice undergoing BMNC therapy were found to have normalized antibody repertoire and improved cardiac performance measured by ECG, treadmill exercise time and echocardiography. After functional testing, gene expression profiles in cardiac tissue were evaluated using high-density oligonucleotide arrays. Expression of more than 18% of the 11981 quantified unigenes was significantly altered in the infarcted hearts. BMNC therapy restored expression of 2099 (96.2%) of the genes that were altered by infarction but led to altered expression of 286 other genes, considered to be a side effect of the treatment. Transcriptional therapeutic efficacy, a metric calculated using a formula that incorporates both recovery and side effect of treatment, was 73%. In conclusion, our results confirm a beneficial role for bone marrow-derived cell therapy and provide new information on molecular mechanisms operating after BMNC transplantation on post ischemic heart failure in mice. We compared RNA samples extracted from whole hearts of infarcted mouse myocardium treated with bone marrow mononuclear cells control with untreated infarcted and control mice samples by analyzing hybridization to AECOM 32k mouse microarrays (http://microarray1k.aecom.yu.edu/) spotted with Operon version 3.0 70-mer oligonucleotides. The hybridization protocol and the slide type were uniform throughout the entire experiment to minimize the technical noise. Treated, control (sham) and infarcted red-labeled heart samples were hybridized against an in-house prepared green-labeled universal mouse reference.
Project description: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:Background: We have found that extracellular vesicles (EV) secreted by embryonic stem cell-derived cardiovascular progenitor cells (hES-CPg) recapitulate the therapeutic effects of these cells in a model of chronic heart failure (CHF). Objectives: Our goal was to test other cellular sources of EV and to explore their mechanism of action.