The role of shear induced endothelial ZBTB46 in atherosclerosis

剪切诱导内皮ZBTB46在动脉粥样硬化中的作用

基本信息

批准号：
10630214
负责人：
Amir Rezvan
金额：
$ 39万
依托单位：
EMORY UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-07-01 至 2025-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10630214
关键词：
Address Adhesions Affect American Heart Association Apolipoprotein E Apoptosis Apoptotic Area Arteries Atherosclerosis Binding Binding Sites Biological Assay Cardiovascular Physiology Cell Proliferation Cell physiology Cessation of life ChIP-seq Chest Pain Coronary Arteriosclerosis Coronary heart disease Data Dendritic Cells Disease Endothelial Cell Inhibitor Endothelial Cells Endothelium Experimental Models Exposure to Funding Future Gene Expression Gene Targeting Genes Heart failure Hydrogen Peroxide Inflammation Inhibition of Cell Proliferation Knockout Mice Ligation Luciferases Mediating Methods Modeling Morbidity - disease rate Mus Myocardial Infarction Peripheral arterial disease Predisposition Process Proliferating Proteins Public Health Regulation Reporter Role Site Small Interfering RNA Smooth Muscle Myocytes Stimulus Stroke TNF gene Testing Time United States United States National Institutes of Health WNT Signaling Pathway adenoviral mediated angiogenesis athero susceptible atheroprotective comparison control differential expression gain of function immune cell infiltrate in vivo inhibitor insight loss of function monocyte mortality mouse model neovascularization novel overexpression prevent promoter response senescence statistics therapeutic target transcription factor tumor

项目摘要

Project Summary Atherosclerosis is a significant cause of morbidity and mortality in the world, contributing to diseases such as coronary artery disease, peripheral arterial disease, and stroke. It is known that atherosclerosis has a focal propensity to areas of disturbed flow (d-flow) while areas of steady laminar flow (s-flow) are relatively protected. Endothelial cell (EC) activation by d-flow is an early step in atherosclerosis, and it is known that ECs exposed to d-flow show higher levels of inflammation, proliferation, apoptosis, and senescence. We have recently shown that ZBTB46 is a flow regulated transcription factor in ECs, is down-regulated by d-flow, and that its overexpression in ECs inhibits proliferation. While the role of ZBTB46 in atherosclerosis is not known, our preliminary studies suggest that ZBTB46 regulates a number of genes in ECs potentially involved in atherosclerosis, including P21 and SFRP2. We propose to test the overall hypothesis that decreased ZBTB46 expression in ECs in response to d-flow leads to downstream changes in EC gene expression including decreased P21 and SFRP2 gene expression, promoting EC activation, apoptosis, and senescence, and consequently a focal predisposition to atherosclerosis. We address three specific aims for this project: 1) Determine the downstream targets of ZBTB46 involved in flow regulated EC proliferation. First by combining loss of function and gain of function methods and various flow conditions, we will determine if P21 and/or SFRP2 are responsible for the inhibitory effect of ZBTB46 on EC proliferation seen under s-flow. We will also test if P21 and SFRP2 are direct or indirect targets of ZBTB46. Finally, to identify other potential targets of ZBTB46 as a transcription factor, we will use ChIP-seq in ECs expressing ZBTB46, and use the data to test the role of other potential downstream targets of ZBTB46 that could be involved in atherosclerosis in the future. 2) Determine the role of flow regulated ZBTB46 on EC apoptosis, senescence, and inflammation. We will test if EC-ZBTB46 expression affects these aspects of EC function which are seen in atheroprone areas exposed to d-flow, by using siRNA and adenovirus mediated overexpression of ZBTB46 under s-flow and d-flow conditions in conjunction with relevant stimuli such as hydrogen peroxide and TNFa. 3) Determine the effect of EC-ZBTB46 on atherosclerosis in vivo. As ZBTB46 is also expressed in classical dendritic cells which are known to affect atherosclerosis, our team has developed hyperlipidemic EC-targeted ZBTB46-KO mice. Using these mice in experimental models of atherosclerosis, we will determine if loss of ZBTB46 specifically in EC contributes to the flow mediated susceptibility to atherosclerosis by promoting EC apoptosis, senescence, and/or inflammation. Successful completion of our aims will potentially find a novel regulator of EC functions contributing to atherosclerosis and may identify novel potential therapeutic targets for atherosclerosis.

项目摘要动脉粥样硬化是世界上发病率和死亡率的重要原因，导致诸如冠状动脉疾病，周围动脉疾病和中风。众所周知，动脉粥样硬化具有局灶性对流动障碍区域（D-Flow）区域的倾向，而稳定层流（S-Flow）的区域受到相对保护。 D-Flow激活内皮细胞（EC）是动脉粥样硬化的早期一步，众所周知，EC暴露于 d流动显示较高的炎症，增殖，凋亡和衰老。我们最近显示了 ZBTB46是EC中的流动调节因子，被D-Flow降低，并且它 EC中的过表达抑制增殖。虽然ZBTB46在动脉粥样硬化中的作用尚不清楚，但我们初步研究表明，ZBTB46调节了潜在参与的EC中的许多基因动脉粥样硬化，包括P21和SFRP2。我们建议测试降低ZBTB46的总体假设响应于D流的EC中的表达导致EC基因表达的下游变化，包括 p21和SFRP2基因表达降低，促进EC激活，凋亡和衰老，以及因此，对动脉粥样硬化的局灶性倾向。我们针对该项目的三个具体目标：1）确定与流动调节EC增殖有关的ZBTB46的下游靶标。首先结合损失功能和功能方法的增益和各种流动条件，我们将确定P21和/或SFRP2是否为负责ZBTB46对S-Flow下的EC增殖的抑制作用。我们还将测试P21和 SFRP2是ZBTB46的直接或间接目标。最后，确定ZBTB46的其他潜在目标转录因子，我们将在表达ZBTB46的EC中使用chip-seq，并使用数据测试其他 ZBTB46的潜在下游靶标可能会参与动脉粥样硬化。 2）确定流动调节ZBTB46在EC凋亡，衰老和炎症中的作用。我们将测试EC-ZBTB46是否表达会影响EC功能的这些方面，通过使用 siRNA和腺病毒介导的ZBTB46在S-Flow和D-Flow条件下的过表达与相关的刺激，例如过氧化氢和TNFA。 3）确定EC-ZBTB46对动脉粥样硬化在体内。由于ZBTB46在经典的树突状细胞中也表达，已知会影响动脉粥样硬化，我们的团队已经开发了高脂性EC靶向的ZBTB46-KO小鼠。在动脉粥样硬化的实验模型，我们将确定ZBTB46的损失是否特别有助于通过促进EC凋亡，衰老和/或炎症，流动介导的动脉粥样硬化易感性。成功完成我们的目标将有可能找到促成EC功能的新型调节剂动脉粥样硬化，可能鉴定出动脉粥样硬化的新型潜在治疗靶标。