Resolution of inflammation in healing myocardial infarcts

缓解心肌梗塞中的炎症

• 批准号: 10364949
• 负责人: Nikolaos G Frangogiannis
• 金额: $ 70.94万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-01-15 至 2025-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10364949
• 关键词: Activins; Acute; Affect; Anti-Inflammatory Agents; Attenuated; Binding; Biological Assay; Blood Vessels; Bone Morphogenetic Proteins; CXCL12 gene; Cardiac; Cardiac Myocytes; Cell Nucleus; Cells; Characteristics; Cicatrix; Complex; Cytokine Activation; Data; Development; Differentiation and Growth; Down-Regulation; Endothelial Cells; Event; Family; Fibroblasts; Fibrosis; Flow Cytometry; Follistatin; Functional disorder; Geometry; Growth Factor; Heart; Heart failure; Heterogeneity; Hypertrophy; Immune; In Vitro; Infarction; Inflammation; Inflammatory Response; Integrins; Knockout Mice; Ligand Binding; Mediating; Molecular; Myeloid Cells; Myocardial; Myocardial Infarction; Myocardium; Myofibroblast; Pathway interactions; Patients; Pericytes; Phagocytes; Pharmacology; Phenotype; Play; Property; Protein Isoforms; Proteomics; Reactive Oxygen Species; Receptor Activation; Regulation; Reperfusion Therapy; Resolution; Role; STAT6 gene; Signal Transduction; TGF-beta type I receptor; TGFB1 gene; TGFBR2 gene; TP53 gene; Testing; Time; Transforming Growth Factor beta; Transforming Growth Factor beta Receptors; angiogenesis; cardiac repair; chemokine; experimental study; gain of function; healing; immune function; in vitro Assay; in vivo; insight; interstitial; loss of function; macrophage; member; morphogens; mortality; receptor; recruit; repair function; repaired; response; single-cell RNA sequencing; transcriptome sequencing; transcriptomics; transforming growth factor beta3

ABSTRACT: The members of the Transforming Growth Factor (TGF)-Beta superfamily (TGF-Betas, Bone Morphogenetic Proteins/BMPs, Growth differentiation factors/GDFs, activins, etc), play a central role in repair, remodeling and fibrosis of the infarcted heart. TGF-Betas signal through heterotetrameric complexes composed of two type II and two type I TGF-Beta receptors (TBetaRII and TBetaRI respectively). TBetaRI activation transduces signals through a family of intracellular effectors, the receptor-activated Smads (R-Smads: Smad1/2/3/5/8), or by stimulating non-Smad cascades. The profile of type 1 receptors activated by a specific TGF-Beta superfamily member determines which downstream signaling cascade will be activated. Traditional concepts suggest that the 3 TGF-Beta isoforms (TGF-Beta1, -Beta2 and -Beta3) signal through Smad2 or Smad3, whereas BMPs activate Smad1 and Smad5. Studies in endothelial cells have challenged this concept, suggesting that TGF-Betas may stimulate both Smad1 and Smad2/3 signaling, with the 2 pathways exerting antagonistic effects. We have previously demonstrated cell- specific actions of TGF-Beta/Smad3 signaling in myocardial infarction that contribute to myofibroblast activation, modulate cardiomyocyte survival and function, and regulate macrophage phagocytic activity, and anti-inflammatory transition. However, the role of the Smad1 cascade in repair, remodeling and fibrosis of the infarcted heart remains unknown, and the effects of Smad1 on phenotype and function of immune and reparative cells has not been investigated. Our proposal explores the role of Smad1 signaling in repair and remodeling of the infarcted heart. Our preliminary data show that Smad1 is activated predominantly in infarct myofibroblasts and macrophages, but also in border zone cardiomyocytes and in pericytes. In vitro, TGF-Beta isoforms potently activate Smad1 in cardiac reparative cells (macrophages and fibroblasts). In vivo, our cell-specific loss-of-function experiments, revealed unanticipated anti-fibrotic effects of Smad1 in myofibroblasts and showed that in myeloid cells, Smad1 regulates the angiogenic properties of macrophages. Accordingly, we will explore cell-specific actions of Smad1 in cardiac repair and we will dissect the molecular mechanisms of Smad1 effects in 4 specific aims: Specific aim 1: to investigate the role of Smad1 in regulation of fibroblast phenotype following MI and to explore the molecular basis for Smad1-mediated anti-fibrotic effects. RNA- seq analysis suggests that the antifibrotic effects of Smad1 may involve competition with Smad3 signaling, or interactions with STAT6 and p53 fibrogenic pathways. The role of these mechanisms will be investigated in vitro and in vivo. Specific aim 2: to study the role of macrophage Smad1 in repair and remodeling of the infarcted heart. Our preliminary data show that macrophage-specific Smad1 perturbs repair of the infarcted heart by restraining the angiogenic properties of macrophages through downregulation of the angiogenic chemokine CXCL12. We will explore the mechanisms for the angiogenesis-regulating actions of macrophage Smad1 in vitro and in vivo. Specific aim 3: to investigate the molecular basis for Smad1 vs Smad2/3 activation in fibroblasts and macrophages and to test the hypothesis that the TBetaRI profile may be a key determinant of the transcriptomic and functional heterogeneity of reparative cells. We will use flow cytometry and single nucleus RNA-seq in vivo, and cell biological assays in vitro, to study the role of specific TBetaRIs in regulation of the phenotype and functional properties of fibroblasts and macrophages. Specific aim 4: to study the role of the Smad1 pathway in cardiomyocytes and in pericytes in the infarcted and remodeling myocardium. The proposed studies will provide for the first time insights into the role of Smad1 in regulation of myocardial inflammation and fibrosis, will dissect the mechanisms for Smad1-mediated actions, and will explore the molecular basis for the functional and transcriptomic heterogeneity of immune and reparative cells in the infarcted myocardium.

摘要： 转化生长因子-β超家族(转化生长因子-β，骨 形态发生蛋白/骨形态发生蛋白、生长分化因子/GDF、激活素等)，在 心肌梗死的修复、重塑和纤维化。转化生长因子-β通过异四聚体传递信号 由两个II型和两个I型转化生长因子-β受体(TBetaRII和TBetaRI)组成的复合体。TBetaRI 激活通过一系列细胞内效应器，即受体激活的Smads来传递信号 (R-SMADS：SMAD1/2/3/5/8)，或通过刺激非SMAD级联。1型受体的研究概况 由特定的转化生长因子-β超家族成员激活决定了哪个下游信号级联 将被激活。传统概念认为，3种转化生长因子-β亚型(转化生长因子-β1、-β2和-β3)是信号转导途径。 通过Smad2或Smad3，而BMP激活Smad1和Smad5。关于内皮细胞的研究 都对这一概念提出了挑战，表明转化生长因子-β可能同时刺激Smad1和Smad2/3 信号转导，两条通路发挥拮抗作用。我们之前已经演示过细胞- 转化生长因子-β/Smad3信号在心肌梗死形成肌成纤维细胞中的特殊作用 激活，调节心肌细胞的存活和功能，调节巨噬细胞的吞噬活性， 和抗炎过渡。然而，Smad1级联在修复、重塑和 心肌梗死后的纤维化机制尚不清楚，Smad1对心肌纤维化的表型和功能的影响尚不清楚。 免疫和修复细胞还没有被研究过。 我们的建议探讨了Smad1信号在细胞修复和重塑中的作用 心脏梗死。我们的初步数据显示，Smad1主要在脑梗塞中被激活 肌成纤维细胞和巨噬细胞，也存在于交界区的心肌细胞和周细胞。在体外， 转化生长因子-β亚型可有效激活心脏修复细胞(巨噬细胞和成纤维细胞)中的Smad1。在……里面 体内，我们的细胞特异性功能丧失实验，揭示了Smad1意想不到的抗纤维化作用 研究表明，在髓系细胞中，Smad1调节血管生成特性。 巨噬细胞。因此，我们将探索Smad1在心脏修复中的细胞特异性作用，我们将 从四个方面剖析Smad1效应的分子机制： 特异性目的1：探讨Smad1在成纤维细胞表型调控中的作用 并探讨Smad1介导的抗纤维化作用的分子基础。RNA- 序列分析表明，Smad1的抗肝纤维化作用可能与Smad3竞争有关 信号转导，或与STAT6和P53纤维化通路的相互作用。这些机制的作用将是 在体外和体内进行研究。 特异性目的2：研究巨噬细胞Smad1在血管修复和重塑中的作用 心脏梗死。我们的初步数据显示，巨噬细胞特异性的Smad1干扰了 通过下调巨噬细胞的血管生成抑制心肌梗死 血管生成趋化因子CXCL12。我们将探讨血管生成调控的机制。 巨噬细胞Smad1的体内外作用。 特异性目标3：研究Smad1和Smad2/3激活的分子基础 成纤维细胞和巨噬细胞，并检验TBetaRI谱可能是关键的假设 修复细胞转录和功能异质性的决定因素。我们将使用 流式细胞术和体内单核rna-seq，以及体外细胞生物学检测，以研究其作用。 特异性TBetaRis对成纤维细胞表型和功能特性的调节 巨噬细胞。 特异性目标4：研究Smad1通路在心肌细胞和心肌细胞中的作用 梗塞和重塑心肌中的周细胞。 拟议的研究将首次深入了解Smad1在调控中的作用 将剖析Smad1介导的作用机制，以及 将探索免疫和转录异质性的分子基础 梗死心肌中的修复细胞。