Ectonucleotidases in ischemic heart disease

美国每年有100多万人患有心肌梗塞(MI)。心肌 心肌梗死(MI)引起强烈的炎症反应。旁分泌和自分泌释放胞外ATP (EATP)，通过嘌呤能P2受体(P2X和P2Y)传递信号，是巨噬细胞和 成纤维细胞功能。胞外ATP被跨膜蛋白胞外核苷三磷酸水解酶 二磷酸水解酶1(CD39)产生一磷酸腺苷(AMP)，从而停止促炎作用 ATP介导的信号转导。该项目的目标是确定巨噬细胞和成纤维细胞的作用- CD39在心肌梗死后心脏修复中的表达。为了实现这一目标，我们提出以下建议：目标1： 确定CD39上调是否是抑制自分泌ATP驱动的保护性机制 炎症，防止过度的巨噬细胞反应，并防止不利的心肌梗死后纤维化 将：1)确定CD39活性如何影响TLR-4依赖和IL-4依赖的巨噬细胞功能 体外，2)测定CD39活性对体内巨噬细胞表型转化的影响 3)确定巨噬细胞CD39在心肌梗死后修复中的作用。 心肌梗死修复后体内纤维化。目的2：分析参与CD39抑制的嘌呤能通路。 转化生长因子-CD391对心脏成纤维细胞的激活作用及β上调对成纤维细胞的调控作用 功能和调节心肌梗死后的心脏修复我们将1)确定下游靶点，通过 CD39减轻转化生长因子-β1介导的心脏成纤维细胞炎症和纤维化反应，2)测定 CD39在心脏修复过程中对在体心脏成纤维细胞表型的影响 成纤维细胞CD39对心肌梗死后修复过程中细胞外基质重塑的影响。次要目标：确定 CD39表达对细胞外基质重塑的影响我们将使用最先进的MALDI成像 质谱学研究心肌梗死后早期和晚期心肌细胞外基质重塑的生物特征 确定CD39在ECM重塑中的作用。这些研究的结果将揭示 CD39调节心肌梗死后心肌修复的途径及其可能提供新的治疗方法 不仅可以治疗心脏纤维化，还可以治疗皮肤、肺、骨髓、肝脏或肾脏， 从而提供了重要的翻译影响。

More than 1 million people in the United States suffer a myocardial infarction (MI) each year. Myocardial infarction (MI) evokes an intense inflammatory response. Paracrine and autocrine released extracellular ATP (eATP), signaling through purinergic P2 receptors (P2X and P2Y), is a potent modulator of macrophage and fibroblast function. Extracellular ATP is hydrolyzed by the transmembrane protein Ectonucleoside triphosphate diphosphohydrolase 1 (CD39) to yield adenosine monophosphate (AMP), thereby halting pro-inflammatory ATP-mediated signaling. This goal of the project is to define the role of the macrophage and fibroblast- expressed CD39 on post-MI cardiac repair. To accomplish this goal we propose the following: Aim 1: To determine whether CD39 up-regulation is a protective mechanism that constrains autocrine ATP-driven inflammation, preventing exaggerated macrophage responses, and protecting from adverse post-MI fibrosis we will: 1) determine how CD39 activity impacts TLR-4-dependent and IL-4-dependent macrophage functions in vitro, 2) determine the impact of CD39 activity on the in vivo macrophage phenotypic transition from inflammatory to profibrotic functions during post-MI repair, and 3) Determine the role of macrophage CD39 on post-MI repair fibrosis in vivo. Aim 2: To dissect the purinergic pathways involved in CD39-mediated restraint of TGF-β1 activation of cardiac fibroblasts and determine the role of CD39 upregulation in modulating fibroblast function and regulating cardiac repair following MI we will 1) determine the downstream targets through which CD39 attenuates TGF-β1-mediated cardiac fibroblast inflammatory and fibrotic responses, 2) determine the impact of CD39 on the in vivo cardiac fibroblast phenotype during cardiac repair and 3) determine the impact of fibroblast CD39 on in vivo extracellular matrix remodeling during post-MI repair. Secondary Aim: To determine the impact of CD39 expression extracellular matrix remodeling we will use state-of-the-art MALDI-imaging mass spectroscopy to define the early and late biosignatures of cardiac ECM remodeling post-MI and determine the impact of CD39 on ECM remodeling. The outcomes of these studies will reveal the fundamental pathways by which CD39 regulates post-MI myocardial repair and could allow novel therapeutic approaches not only to treat fibrotic disorders of the heart, but also of the skin, lungs, bone marrow, liver, or kidneys, thereby providing an important translational impact.