The pluripotency factor OCT4 plays an athero-protective role in endothelial cells

多能因子 OCT4 在内皮细胞中发挥动脉粥样硬化保护作用

基本信息

批准号：
10266228
负责人：
Olga Cherepanova
金额：
$ 40.25万
依托单位：
CLEVELAND CLINIC LERNER COM-CWRU
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：
2020-09-30 至 2022-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10266228
关键词：
ABCG2 gene ATP-Binding Cassette Transporters Adhesions Aorta ApoE knockout mouse Apolipoprotein E Arterial Fatty Streak Arteries Atherosclerosis Attenuated Binding Blood Vessels Brain Cell Adhesion Molecules Cell Lineage Cells Cessation of life ChIP-seq Cholesterol Chronic Disease Clinical Cluster Analysis Coronary artery Data Development Disease Endothelial Cells Endothelium Functional disorder Gatekeeping Gene Expression Profiling Genes Goals Growth Heme High Fat Diet Human Impairment In Vitro Inflammation Inflammatory Inflammatory Response Knock-out Lead Lesion Lipids Lung Mediating Mesenchymal Metabolic Metabolism Modification Molecular Mus Myocardial Myocardial Infarction NF-kappa B Oxidative Stress Pathogenesis Phenotype Play Population Heterogeneity Porphyrins Production Proteins Pulmonary artery structure Role Signal Transduction Site-Directed Mutagenesis Smooth Muscle Myocytes Somatic Cell Specimen Testing Therapeutic Time Tissue-Specific Gene Expression Up-Regulation Vascular Cell Adhesion Molecule-1 Vascular Smooth Muscle Xenobiotics atherogenesis atheroprotective base cardiac repair cell injury chromatin immunoprecipitation cytokine embryonic stem cell gene repression in vivo indexing induced pluripotent stem cell intercellular cell adhesion molecule macrophage monocyte mouse model neuroinflammation novel novel therapeutic intervention oxidized lipid pluripotency prevent promoter recruit response single-cell RNA sequencing stem cell division transcriptome western diet

项目摘要

ABSTRACT Atherosclerosis is a chronic disease of the arterial wall which contributes to >40% of all deaths in the USA. Although previous studies provide clear evidence for the importance of endothelial cell (EC) activation and phenotypic modulation during atherosclerosis, little is known about key regulatory mechanisms for EC plasticity that influence lesion growth and/or stability. Recently, we found that the embryonic stem cell/iPS factor OCT4 that was believed to be silenced in somatic cells plays an athero-protective role in smooth muscle cells (SMC) by regulating SMC phenotypic transitions. We then asked if OCT4 might also play a key role in regulating plasticity of EC during atherosclerosis development. Our preliminary results using a novel EC-specific Oct4 knockout Apoe-/- mouse model demonstrated that loss of Oct4 in EC increases the total lipid burden within aortas and occurrence of plaques within pulmonary arteries of Apoe-/- mice fed a long-term high-fat diet, indicating that OCT4 plays a key functional role within EC. To further understand molecular mechanisms whereby EC-derived OCT4 inhibits the accumulation of lipids we performed a single cell (sc)RNA-seq analysis on the aortic and lung EC sorted from EC-Oct4 knockout and wild-type Apoe-/- mice fed a Western diet for five weeks. Differential gene expression analyses identified 405 (aorta), and 250 (lung) significantly dysregulated genes in Oct4 knockout EC as compared to wild type EC, including upregulation of genes involved into Endothelial-to-Mesenchymal transition (Endo-MT), pro-inflammatory and adhesion molecules, and down-regulation of genes involved into xenobiotic and heme metabolism. Also, using results from the in vivo OCT4 ChIP-seq, we found that one of the genes significantly down-regulated in the Oct4 knockout lung and aortic EC, Abcg2, is a putative target of OCT4. ABCG2 is an ATP-binding cassette transporter responsible for extruding toxic xenobiotics, including heme from cells. Previous studies demonstrated that EC-derived ABCG2 plays a pivotal role in cardiac repair after myocardial infarction and protective role against oxidative stress and neuroinflammatory response in mouse brains by inhibiting NF-kB-signaling. However, there is no data about the potential protective role of ABCG2 in EC during atherosclerosis development. Our overall hypothesis is that the pluripotency factor OCT4 plays a critical athero-protective role in EC during the development of atherosclerosis through activation of the gatekeeper protein ABCG2, an adaptive mechanism to decrease oxidative stress and protect EC from ROS- induced activation and inflammatory response. Aim 1 will test the hypothesis that OCT4 plays an athero- protective role by preventing EC from activation and phenotypic modification at the early and late stages of atherosclerosis. Aim 2 will test the hypothesis that OCT4 attenuates EC-activation and EC inflammatory response at least in part through up-regulating Abcg2 that protects EC from oxidative stress by decreasing intracellular heme levels and ROS-mediated NF-kB-dependent EC inflammatory response. These studies will lead to an identification of novel cell-specific therapeutic approaches for treating or preventing atherosclerosis.

抽象的动脉粥样硬化是一种动脉壁的慢性疾病，占美国所有死亡人数的40％。尽管以前的研究为内皮细胞（EC）激活的重要性提供了明确的证据在动脉粥样硬化期间的表型调节，关于EC可塑性的关键调节机制知之甚少这会影响病变的生长和/或稳定性。最近，我们发现胚胎干细胞/IPS因子OCT4 据信在体细胞中被沉默的这种情况在平滑肌细胞（SMC）中扮演动脉化作用通过调节SMC表型过渡。然后，我们询问Oct4是否也可能在调节中起关键作用动脉粥样硬化发展期间EC的可塑性。我们使用新型EC特异性OCT4的初步结果敲除apoe - / - 鼠标模型表明，EC中OCT4的损失增加了主动脉内的总脂质负担并在ApoE - / - 小鼠的肺动脉内发生斑块 - 喂养长期高脂饮食，表明 OCT4在EC中起关键功能作用。为了进一步了解分子机制 OCT4抑制了脂质的积累，我们对主动脉和肺进行了单个细胞（SC）RNA-Seq分析 EC从EC-OCT4敲除和野生型APOE - / - 小鼠喂养西方饮食五周。差异基因表达分析鉴定出405（主动脉）和250（肺）在OCT4敲除EC中显着失调的基因失调与野生型EC相比，包括在内皮到间质涉及的基因上调过渡（内托-MT），促炎和粘附分子以及涉及的基因下调异种生物和血红素代谢。此外，使用体内Oct4芯片序列的结果，我们发现其中一个 OCT4基因敲除肺和主动脉EC ABCG2中明显下调的基因是OCT4的推定靶标。 ABCG2是一种ATP结合盒转运蛋白，负责挤出有毒的异种生物，包括血红素。细胞。先前的研究表明，EC衍生的ABCG2在心脏修复中起关键作用心肌梗死和保护小鼠氧化应激和神经炎症反应的保护作用大脑通过抑制NF-KB信号。但是，没有关于ABCG2潜在保护作用的数据动脉粥样硬化发展期间的EC。我们的总体假设是多能因素OCT4扮演通过激活而在动脉粥样硬化发展期间，关键的动脉粥样硬化作用在EC中网守蛋白ABCG2，一种自适应机制，可降低氧化应激并保护EC免受ROS- 诱导的激活和炎症反应。 AIM 1将检验Oct4扮演动脉粥样硬化的假设通过防止EC在早期和晚期的激活和表型修饰来进行保护作用动脉粥样硬化。 AIM 2将检验OCT4减弱EC激活和EC炎症的假设至少部分通过上调ABCG2的反应，该ABCG2通过减少来保护EC免受氧化应激细胞内血红素水平和ROS介导的NF-KB依赖性EC炎症反应。这些研究会导致对治疗或预防动脉粥样硬化的新细胞特异性治疗方法的鉴定。