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.

项目概要动脉粥样硬化是世界范围内发病和死亡的一个重要原因，导致以下疾病冠状动脉疾病、外周动脉疾病和中风。众所周知，动脉粥样硬化具有局灶性扰动流（d 流）区域的倾向，而稳定层流（s 流）区域则相对受到保护。 d-flow 激活内皮细胞 (EC) 是动脉粥样硬化的早期步骤，众所周知，EC 暴露于 d-flow 显示更高水平的炎症、增殖、细胞凋亡和衰老。我们最近展示了 ZBTB46 是 EC 中的流量调节转录因子，被 d-flow 下调，并且其 EC 中的过度表达会抑制增殖。虽然 ZBTB46 在动脉粥样硬化中的作用尚不清楚，但我们的研究初步研究表明，ZBTB46 调节 EC 中的许多基因，可能参与动脉粥样硬化，包括 P21 和 SFRP2。我们建议检验减少 ZBTB46 的总体假设 EC 中响应 d-flow 的表达导致 EC 基因表达的下游变化，包括降低 P21 和 SFRP2 基因表达，促进 EC 激活、细胞凋亡和衰老，以及因此，局部易患动脉粥样硬化。我们针对该项目提出了三个具体目标：1) 确定 ZBTB46 参与流量调节 EC 增殖的下游靶标。首先通过组合损失函数方法和增益函数方法以及各种流量条件，我们将确定 P21 和/或 SFRP2 是否负责 ZBTB46 对 s-flow 下 EC 增殖的抑制作用。我们还将测试 P21 和 SFRP2 是 ZBTB46 的直接或间接靶标。最后，确定 ZBTB46 的其他潜在目标作为转录因子，我们将在表达 ZBTB46 的 EC 中使用 ChIP-seq，并使用数据来测试其他转录因子的作用 ZBTB46 的潜在下游靶标未来可能参与动脉粥样硬化。 2）确定流量调节的 ZBTB46 对 EC 细胞凋亡、衰老和炎症的作用。我们将测试EC-ZBTB46是否表达影响 EC 功能的这些方面，这些方面在暴露于 d-flow 的动脉粥样硬化易发区域中可见，通过使用 s-flow和d-flow条件下siRNA和腺病毒联合介导ZBTB46的过表达与相关刺激，如过氧化氢和 TNFa。 3) 确定EC-ZBTB46对体内动脉粥样硬化。由于 ZBTB46 也在经典树突状细胞中表达，已知其会影响针对动脉粥样硬化，我们团队开发了高脂血症 EC 靶向 ZBTB46-KO 小鼠。使用这些小鼠动脉粥样硬化的实验模型中，我们将确定 EC 中 ZBTB46 的丢失是否会导致血流通过促进 EC 凋亡、衰老和/或炎症来介导动脉粥样硬化的易感性。成功完成我们的目标将有可能找到一种新的 EC 功能调节器，有助于动脉粥样硬化，并可能确定动脉粥样硬化的新的潜在治疗靶点。