Epigenetic-mediated Notch pathway activation promotes elastin aortopathy

表观遗传介导的Notch通路激活促进弹性蛋白主动脉病

基本信息

批准号：
10595308
负责人：
Daniel Greif
金额：
$ 65.47万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-02-01 至 2028-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10595308
关键词：
Aorta Aortic Valve Stenosis Arterial Occlusive Diseases Arteries Attenuated Birth Blood flow Cardiovascular Diseases Cell Culture Techniques Cell Differentiation process Cell Nucleus Cell Proliferation Cell Separation Cells Cellular biology Chromatin Coupled DNA Methylation DNA Modification Methylases Data Development Disease Elasticity Elastin Embryo Enzymes Epigenetic Process Gene Expression Genes Genetic Diseases Genomics HDAC1 gene Heart Heart failure Histone Acetylation Histone Deacetylase Human Kinetics Left Ligands Link Loss of Heterozygosity Mediating Mediator Medical Modification Molecular Mus Muscle Cells Mutant Strains Mice NOTCH3 gene Obstruction Operative Surgical Procedures Paper Pathogenesis Pathology Pathway interactions Patients Play Proliferating Regulator Genes Reporting Repression Risk Role Sampling Signal Transduction Smooth Muscle Myocytes Stenosis Sudden Death Supravalvular aortic stenosis System Techniques Testing Tissue Sample Tissues Transgenic Mice Tropoelastin Vascular Diseases Williams Syndrome attenuation cell behavior chromatin remodeling developmental genetics epigenetic regulation gamma secretase gene repression genome-wide in vivo induced pluripotent stem cell loss of function mutation mouse model mutant neonatal mice notch protein novel novel therapeutic intervention pup receptor therapeutic target transcriptomics

项目摘要

PROJECT SUMMARY Supravalvular aortic stenosis (SVAS) is a developmental cardiovascular disease, occurring alone or in Williams Beuren Syndrome (WBS), and results in excessive arterial smooth muscle cell (SMC) proliferation and lumen obstruction. Our long-term objective is to elucidate how this pathology can be attenuated. Heterozygous loss-of-function mutations or deletions of the elastin gene ELN cause SVAS. Eln(-/-) embryos and Eln(+/-) neonatal mice develop arterial disease with features similar to human SVAS4-6. Elastin forms the major component of the elastic lamellae in arteries. Defective lamellae are associated with excessive developmental SMC proliferation in SVAS. If untreated, SVAS results in heart failure and an increased risk of sudden death, and major surgery, which carries substantial risk, is the only treatment. Medical therapies are lacking because mechanisms linking defective elastin and SMC hypermuscularization are incompletely defined. We aim to elucidate molecular and cellular mechanisms underlying elastin aortopathy. The Notch pathway is critical in regulating SMC biology, and we recently reported a role for Notch in SVAS pathogenesis (JCI, 2022). Signaling via Notch ligand Jagged1 (JAG1) and NOTCH3 receptor in SMCs activates proliferation. Our studies reveal that JAG1-NOTCH3 pathway components are upregulated after elastin depletion. Importantly, we determined that inhibiting the NOTCH3 pathway in Eln(-/-) embryos attenuates aortic hypermuscularization and stenosis and reverses established hypermuscularization in Eln(+/-) pups. Epigenetic modifications influence gene expression by altering chromatin accessibility but prior to our JCI paper, have not been explored in elastin deficiency. Our initial data indicate that elastin deficient aortas and SMCs display reduced DNA methylation, elevated histone acetylation and reduced expression of DNA methyltransferase 1 (DNMT1) and histone deacetylase 1 (HDAC1). We hypothesize that elastin deficiency attenuates DNMT1- and HDAC1-mediated repression of Notch pathway genes to promote aortic hypermuscularization and stenosis. The proposed studies use cell culture, mouse models, de-identified human samples and advanced genomic and epigenetic techniques to uncover mechanisms of SVAS that can be therapeutically targeted. We will test our hypothesis in two aims. Aim 1 will determine how elastin deficiency alters epigenetic mediators and the epigenetic landscape in human and murine SMCs, including 1a) identifying elastin-regulated epigenetic enzymes, 1b) determining mechanisms by which elastin deficiency modulates epigenetic regulators, 1c) an integrated genome-wide epigenetic and transcriptomic analysis to identify new regulatory mechanisms and 1d) characterization of epigenetic enzymes and marks in human samples. Aim 2 will elucidate the relationship between elastin, chromatin remodeling and the Notch pathway, including 2a) identifying causal epigenetic mechanisms, 2b) kinetics and 2c) testing HAT inhibition as a therapy in mouse models of SVAS. These studies promise to yield new treatments for this lethal genetic disease.

项目摘要上主动脉狭窄（SVAS）是一种发育性心血管疾病，单独或发生威廉姆斯·贝伦综合症（WBS），导致过度动脉平滑肌细胞（SMC）增殖和管腔阻塞。我们的长期目标是阐明如何减弱这种病理。弹性丧失的突变或弹性蛋白基因的缺失引起SVA。 Eln（ - / - ）胚胎和 ELN（+/-）新生小鼠患有动脉疾病，其特征类似于人类SVAS4-6。弹性蛋白构成专业动脉中弹性薄片的成分。缺陷层与过度发育有关 SVA中的SMC扩散。如果未经治疗，SVA会导致心力衰竭和猝死风险增加，唯一的治疗方法是带有重大风险的大型手术。缺乏医疗疗法，因为连接有缺陷的弹性蛋白和SMC高肌化的机制未完全定义。我们的目标阐明弹性弹性蛋白主动脉蛋白主动脉肿大的分子和细胞机制。 Notch途径对于调节SMC生物学至关重要，我们最近报告了Notch在 SVAS发病机理（JCI，2022年）。通过Notch配体Jagged1（JAG1）和Notch3受体在SMC中发出信号传导激活增殖。我们的研究表明，JAG1-NOTCH3途径成分在弹性蛋白耗竭。重要的是，我们确定抑制ELN（ - / - ）胚胎中的Notch3途径减弱主动脉高肌间化和狭窄和逆转在ELN（+/-）幼犬中建立的高肌化。表观遗传修饰通过改变染色质的可及性来影响基因表达，但在我们之前 JCI纸，尚未在弹性蛋白缺乏症中探索。我们的初始数据表明弹性蛋白缺乏主动脉 SMC表现出降低的DNA甲基化，组蛋白乙酰化升高和DNA表达降低甲基转移酶1（DNMT1）和组蛋白脱乙酰基酶1（HDAC1）。我们假设弹性蛋白缺乏症减弱DNMT1-和HDAC1介导的Notch途径基因的抑制作用以促进主动脉高肌化和狭窄。拟议的研究使用细胞培养，小鼠模型，去识别的人类样品以及先进的基因组和表观遗传技术，以发现可以是SVA的机制治疗目标。我们将以两个目标来检验我们的假设。 AIM 1将确定弹性蛋白缺乏症如何改变人和鼠SMC中的表观遗传介体和表观遗传景观，包括1A）弹性蛋白调节的表观遗传酶，1B）确定弹性蛋白缺乏调节的机制表观遗传调节剂，1C）一种整体基因组的表观遗传学和转录组分析，以鉴定新的人类样品中表观遗传酶和标记的调节机制和1d）表征。目标2 将阐明弹性蛋白，染色质重塑和Notch途径之间的关系，包括2A）鉴定因果表观遗传机制，2B）动力学和2C）测试HAT抑制作用作为小鼠的治疗 SVA的模型。这些研究有望为这种致命的遗传疾病产生新的治疗方法。