CD45-mediated endothelial-to-mesenchymal transition in cardiovascular disease

CD45介导的心血管疾病中的内皮间质转化

• 批准号: 9917371
• 负责人: Hong Chen
• 金额: $ 76.22万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-01-15 至 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9917371
• 关键词: Age; Apolipoprotein E; Arterial Fatty Streak; Atherosclerosis; Attenuated; Automobile Driving; Biochemical; Biological; Biological Process; CRISPR/Cas technology; Cardiovascular Diseases; Cause of Death; Cells; Complement; Coronary heart disease; Data; Development; Diet; Down-Regulation; Endocytosis; Endothelial Cells; Endothelium; Epigenetic Process; Exhibits; Extracellular Matrix Proteins; FGFR1 gene; Female; Fibroblast Growth Factor; Genetic; Goals; HDAC2 gene; Hematopoietic; Human; Hyperplasia; Inflammatory; Intervention; Ischemic Stroke; Knock-out; Knockout Mice; Lesion; Lipids; Mediating; Mesenchymal; MicroRNAs; Mission; Modeling; Molecular; Mus; Myocardial Infarction; PTPRC gene; Pathologic Processes; Pathway interactions; Peripheral arterial disease; Pharmacology; Play; Prevention approach; Prevention program; Regulation; Repression; Research; Resolution; Role; Signal Pathway; Signal Transduction; Stroke; Tamoxifen; Testing; Therapeutic; Translating; United States; United States National Institutes of Health; Up-Regulation; Vascular Diseases; atherogenesis; combat; endothelial dysfunction; epsin; genetic regulatory protein; hemogenic endothelium; in vivo; innovation; male; molecular marker; mortality; mutant; new therapeutic target; novel; overexpression; phosphatase inhibitor; prevent; therapeutic target; transcriptome sequencing; vasoconstriction; western diet

PROJECT SUMMARY/ABSTRACT Atherosclerosis is the primary cause of coronary heart disease, ischemic stroke, and peripheral arterial disease. This progressive vascular disease remains a leading cause of mortality in the United States despite wide-spread use of effective lipid-lowering therapies and prevention programs. While endothelial cell activation and neointimal hyperplasia is a hallmark of the initiation and progression of atherosclerosis, identification of molecular markers of dysfunctional endothelium represents a biological challenge as well as an opportunity to develop new therapeutic targets. In this application, we posit to explore the function of endothelial expressed CD45 in atherosclerosis. CD45 is best known as marker of hematopoietic cells. Our recent data shows that CD45 is indispensable in driving the endothelial-to-mesenchymal transition (EndMT) following myocardial infarction. It is thought that EndMT represents a novel endothelial perturbation and a critical regulator of atherogenesis given that EndMT disrupts the normal function of endothelium by delaminating endothelial cells and giving rise to neointimal mesenchymal cells, which secrete proinflammatory molecules and extracellular matrix proteins that fuel atherosclerosis progression. Whether endothelial CD45 contributes to progression of atherosclerosis by facilitating the EndMT is a completely unknown but highly significant question. In our latest studies, we showed CD45 null mice injected with the atherosclerosis accelerator PCSK9 AAv8 and fed a western diet had a marked reduction in atherogenesis. Conversely, we observed a significant increase in CD45 positive endothelial cells (ECs) that undergo the EndMT in atherosclerotic lesions. We also found that CRISPR/Cas9-mediated activation of CD45 in human ECs induces the EndMT. Intriguingly, CD45 expression is dramatically increased in HDAC2 knockout hemogenic endothelium, suggesting loss of HDAC2-mediated repression of CD45 during EndMT. Further, CD45 expression is altered in miR-155 deficient mice, indicating microRNAs may regulate the expression and function of CD45 during EndMT. Our pilot RNA seq study shows upregulation of epsins and downregulation of KLFs upon CD45 overexpression. Given that epsins promote and KLFs impede atherosclerosis, whether CD45 induces EndMT and promotes atherosclerosis by modulating epsin and KLF function is an entirely novel question. To test this, we will determine the role of endothelial CD45 in regulating atherosclerosis in vivo and interrogate molecular mechanisms 1) by which endothelial CD45 regulates EndMT in atherosclerosis by controlling epsin and KLF expression and 2) underlying how CD45 expression is epigenetically regulated in ECs during EndMT and whether targeting endothelial CD45 promotes atheroma resolution. Our preliminary data presents robust evidence that serves as a strong scientific premise for our proposed study. If successful, our study will not only provide extensive mechanistic evidence for a central role of CD45-mediated signaling in regulating the EndMT, but will provide an innovative approach for the treatment of atherosclerosis and launch a paradigm shift in research to combat cardiovascular disease.

项目概要/摘要 动脉粥样硬化是冠心病、缺血性中风和外周动脉疾病的主要原因 疾病。尽管这种进行性血管疾病仍然是美国死亡的主要原因 广泛使用有效的降脂疗法和预防计划。内皮细胞活化的同时 新生内膜增生是动脉粥样硬化发生和进展的标志， 功能失调的内皮细胞的分子标记既是一种生物学挑战，也是一个机遇 开发新的治疗靶点。在此应用中，我们假设探索内皮细胞表达的功能 CD45在动脉粥样硬化中的作用。 CD45 是最广为人知的造血细胞标记。我们最近的数据表明 CD45 在驱动心肌细胞内皮细胞向间质细胞转化 (EndMT) 过程中不可或缺 梗塞。人们认为 EndMT 代表了一种新的内皮扰动和关键的调节因子 鉴于 EndMT 通过使内皮细胞分层破坏内皮的正常功能，导致动脉粥样硬化 并产生新内膜间充质细胞，分泌促炎分子和细胞外 促进动脉粥样硬化进展的基质蛋白。内皮CD45是否有助于进展 通过促进 EndMT 来预防动脉粥样硬化是一个完全未知但非常重要的问题。在我们最新的 研究中，我们展示了 CD45 缺失小鼠，注射了动脉粥样硬化加速剂 PCSK9 AAv8，并喂食 西方饮食显着减少了动脉粥样硬化的发生。相反，我们观察到 CD45 显着增加 动脉粥样硬化病变中经历 EndMT 的阳性内皮细胞 (EC)。我们还发现 人类 EC 中 CRISPR/Cas9 介导的 CD45 激活可诱导 EndMT。有趣的是，CD45 表达 在 HDAC2 敲除的造血内皮细胞中显着增加，表明 HDAC2 介导的丧失 EndMT 期间抑制 CD45。此外，miR-155 缺陷小鼠中 CD45 表达发生改变，表明 microRNA 可能在 EndMT 过程中调节 CD45 的表达和功能。我们的 RNA seq 试点研究表明 CD45 过表达后，epsins 上调，KLF 下调。鉴于 epsins 促进和 KLFs阻碍动脉粥样硬化，CD45是否诱导EndMT并通过调节促进动脉粥样硬化 epsin 和 KLF 函数是一个全新的问题。为了测试这一点，我们将确定内皮细胞的作用 CD45 在体内调节动脉粥样硬化并探讨分子机制 1) 内皮细胞通过 CD45 通过控制 epsin 和 KLF 表达来调节动脉粥样硬化中的 EndMT，2) 其背后的机制 EndMT 期间 EC 中 CD45 的表达受到表观遗传调控，并且是否靶向内皮 CD45 促进动脉粥样硬化的消退。我们的初步数据提供了强有力的证据，可以作为强有力的科学依据 我们提出的研究的前提。如果成功，我们的研究不仅将提供广泛的机制证据 CD45 介导的信号传导在调节 EndMT 中发挥核心作用，但将提供一种创新方法 用于治疗动脉粥样硬化，并在对抗心血管疾病的研究中发起范式转变。