The role of GATA3-positive macrophages in cardiovascular pathologies

项目总结/摘要 血管炎症是动脉粥样硬化的关键组成部分，其导致斑块不稳定和临床 心血管事件，包括缺血性卒中和心肌梗死（MI）。尽管经过几十年的研究， 导致这些过程的免疫机制在很大程度上仍未得到解决。在骨髓细胞中， 单核细胞/巨噬细胞被认为是心血管疾病发病机制中的主要参与者 （CVD）。新兴的单细胞技术揭示了CVD中的巨噬细胞池是巨大的， 异质的，包含大量转录上不同的亚群。在慢性炎症 在疾病中，GATA 3被认为是T细胞和其他免疫细胞表型的主要调节因子。 然而，GATA 3在巨噬细胞表型调节中的作用是完全未知的。为 我们首次证明GATA 3在巨噬细胞和GATA 3+巨噬细胞中表达， 在急性MI后积聚到心肌中并不利地影响心脏功能。实际上 GATA 3+巨噬细胞的缺乏导致急性MI后心脏功能的显著改善。细胞 培养研究显示IL-33刺激巨噬细胞中GATA 3的表达。我们的长期目标是 探讨CVD中GATA 3+巨噬细胞表型和功能的调控机制。我们 基本原理是，通过鉴定控制GATA 3+巨噬细胞表型的机制， 心血管疾病的发生将提供新的治疗机会。在具体目标1中，我们将利用新的小鼠模型， 结合移植免疫学来检验巨噬细胞特异性IL-33 R信号转导 控制CVD中GATA 3+巨噬细胞的募集和功能。大多数研究旨在描述 人巨噬细胞的表型是在体外进行的;然而， 巨噬细胞在体内是不确定的。GATA 3信号通路在体外人巨噬细胞活性中的作用 在体内是完全未知的。我们已经产生了巨噬细胞特异性的人源化GATA 3， 小鼠作为一种新的临床前平台，以测试人类GATA3信号转导控制 Specific Aim 2中急性MI后巨噬细胞的表型/功能。深入了解动脉粥样硬化 斑块是预防和治疗MI的先决条件。我们发现，GATA 3+巨噬细胞积累到 动脉粥样硬化斑块，巨噬细胞中的GATA 3表达被胆固醇晶体刺激。 然而，GATA 3+巨噬细胞在动脉粥样硬化中的功能完全未知。在具体目标3中， 我们将使用动脉粥样硬化倾向的小鼠模型结合骨髓移植来测试 假设GATA 3+巨噬细胞亚群是致动脉粥样硬化的。这项研究具有创新性，因为 我们将研究GATA3+巨噬细胞在CVD中的作用，这是一个迄今为止尚未研究的过程。

PROJECT SUMMARY/ABSTRACT Vascular inflammation is a key component of atherosclerosis that contributes to plaque instability and clinical cardiovascular events, including ischemic stroke, and myocardial infarction (MI). Despite decades of research, the immune mechanisms contributing to these processes remain largely unresolved. Among myeloid cells, monocytes/macrophages are believed to be major players in the pathogenesis of cardiovascular diseases (CVD). Emerging single-cell technologies have unveiled that the macrophage pool in CVD is vastly heterogeneous, containing a plethora of transcriptionally distinct subpopulations. In chronic inflammatory diseases, GATA3 is regarded as a master regulator of the phenotype of T-cells and other immune cells. However, the role of GATA3 in the regulation of the phenotype of macrophages is completely unknown. For the first time, we demonstrated that GATA3 was expressed in macrophages and GATA3+ macrophages accumulated into the myocardium after acute MI and adversely affected the cardiac function. In fact, the deficiency of GATA3+ macrophages led to a significant improvement of cardiac function after acute MI. Cell culture studies revealed that IL-33 stimulated the expression of GATA3 in macrophages. Our long term goal is to dissect the mechanism/s that regulates the phenotype and function of GATA3+ macrophages in CVD. Our rationale is that by identifying the mechanism/s that control the phenotype of GATA3+ macrophages in the setting of CVD will offer new therapeutic opportunities. In Specific Aim 1, we will utilize novel mouse models in combination with transplant immunology to test the hypothesis that macrophage-specific IL-33R signaling controls the recruitment and function of GATA3+ macrophages in CVD. Most studies aimed at characterizing the phenotypes of human macrophages are performed in vitro; however, the relevance to the function of macrophages in vivo is uncertain. The role of GATA3 signaling in the activity of human macrophages in vitro and in vivo is completely unknown. We have generated macrophage-specific humanized GATA3 sufficient mice as a novel preclinical platform to test the hypothesis that human GATA3 signaling controls the phenotype/function of macrophages after acute MI in Specific Aim 2. In depth knowledge of atherosclerotic plaques is a prerequisite to preventing and treating MI. We found that GATA3+ macrophages accumulated into atherosclerotic plaques, and GATA3 expression in macrophages is stimulated by cholesterol crystals. However, the function of GATA3+ macrophages in atherosclerosis is completely unknown. In Specific Aim 3, we will use atheroma-prone mouse models in combination with bone marrow transplantation to test the hypothesis that GATA3+ macrophage subset/s are atherogenic. The proposed research is innovative because we will investigate the effect of GATA3+ macrophages in CVD, a heretofore-unexamined process.