Endothelial Dysfunction in the Development of Aortic Degeneration, Dissection, and Rupture

主动脉变性、夹层和破裂中的内皮功能障碍

基本信息

批准号：
10662568
负责人：
KETAN B Ghaghada
金额：
$ 66.21万
依托单位：
BAYLOR COLLEGE OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-07-15 至 2026-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10662568
关键词：
Aneurysm Angiotensin II Aorta Aortic Diseases Apoptosis Attention Blood Cardiovascular Diseases Cause of Death Cell Death Cells Cellular Assay Cessation of life Chromatin Chromatin Remodeling Factor DNA DNA Damage Data Development Disease Disease Progression Dissection Elastic Fiber Endothelial Cells Endothelium Enhancers Epigenetic Process Event Extravasation Functional disorder Genes High Fat Diet Human IRF3 gene Image In Vitro Infiltration Inflammation Inflammatory Inflammatory Infiltrate Infusion procedures Interferon Activation Investigation Knockout Mice Life Link Lytic Medial Mediating Microdissection Mission Modeling Molecular Mus National Heart, Lung, and Blood Institute Pathogenesis Pathway interactions Patients Permeability Pharmaceutical Preparations Population Prevention Protein Kinase Interaction Public Health RIPK1 gene Receptor Up-Regulation Research Role Rupture SMARCA4 gene Series Signal Pathway Signal Transduction Smooth Muscle Myocytes Sting Injury Stress TBK1 gene Testing Thoracic Aortic Aneurysm Transposase United States Up-Regulation Wild Type Mouse cell injury contrast enhanced computed tomography endothelial dysfunction epigenomics experimental study in vivo inhibitor insight mouse model nanoparticle novel prevent programs promoter receptor upregulation sensor single-cell RNA sequencing therapeutic target transcription factor transcriptomics

项目摘要

PROJECT SUMMARY Ascending thoracic aortic aneurysms and dissections (ATAAD) are extremely lethal diseases. The contribution of endothelial dysfunction in ATAAD has received limited attention and thus remains poorly understood. Com- pelling evidence from our preliminary studies suggests a critical role for endothelial cell (EC) injury and endo- thelial hyperpermeability in aortic degeneration and ATAAD development. [scRNA-seq analysis of aortas from sporadic ATAAD patients revealed significant EC dysfunction with upregulation of RIP3 and GSDMD that trig- ger necroptosis and pyroptosis, respectively. In a sporadic ATAAD mouse model, aortic challenge with high- fat diet and angiotensin II infusion induced RIP3 and GSDMD in ECs, caused endothelial hyperpermeability, nanoparticle infiltration, interlaminar space expansion, elastic fiber delamination, and microdissection. EC-Rip3- /- or EC-Gsdmd-/- showed markedly reduced nanoparticle infiltration and aortic degeneration.] Further investiga- tion with scATAC-seq showed that aortic challenge increased chromatin accessibility at promoters/enhancers of the pro-inflammatory/pro-death genes, likely by activating pro-inflammatory transcription factors (TFs) such as IRF3. In human ECs, cytosolic DNA and STING-TBK1 signaling was a potent upstream trigger for the acti- vation of IRF3 and BRG1 and induction of GSDMD expression. The objective of this application is to test the central hypothesis that aortic stress activates inflammatory signaling within ECs that triggers epigenetic in- duction of a pro-inflammatory and pro-death program, leading to EC necroptosis/pyroptosis, barrier dysfunc- tion, blood component infiltration, aortic wall degeneration, and ATAAD formation. [Aim 1 will test the hypothe- sis that RIP3-mediated EC necroptosis and GSDMD-mediated EC pyroptosis compromise EC barrier function, thereby facilitating the infiltration of inflammatory blood components and promoting aortic degeneration and ATAAD formation. In our sporadic ATAAD model, we will compare aortic degeneration and ATAAD formation in control mice, EC-Rip3-/- mice, EC-Gsdmd-/- mice, and WT mice treated with a necroptosis/pyroptosis inhibi- tor.] Aim 2 will test the hypothesis that aortic stress activates inflammatory signaling (e.g., STING pathway) within ECs that triggers epigenetic induction of a pro-inflammatory and pro-death program. We will perform scRNA-seq and scATAC-seq analyses of ECs in aortas from unchallenged and challenged WT mice and EC- specific Sting KO mice. Aim 3 will test the hypothesis that stress signals (e.g., cytosolic DNA) trigger sensing- signaling pathways (e.g., STING-TBK) that directly activates chromatin remodeling complexes (e.g., BRG1/SWI/SNF) and TFs (e.g., IRF3) that induce GSDMD expression and promote EC pyroptosis. We will test the hypothesis in cultured human ECs (Aim 3A) and [confirm the mechanistic links in human ATAAD aortas (Aim 3B)]. We expect that the proposed studies will provide novel insight into the molecular mechanisms of sporadic aortic degeneration and ATAAD formation, which will in turn enable development of new treatments for preventing disease progression and its fatal sequelae. 1

项目概要胸主动脉瘤和夹层（ATAAD）是极其致命的疾病。贡献 ATAAD 中内皮功能障碍的研究受到的关注有限，因此仍然知之甚少。康- 我们初步研究的有力证据表明，内皮细胞 (EC) 损伤和内皮细胞损伤具有关键作用。主动脉变性和 ATAAD 发展中的皮细胞通透性过高。 [来自主动脉的 scRNA-seq 分析散发性 ATAAD 患者表现出明显的 EC 功能障碍，RIP3 和 GSDMD 上调，从而触发分别为 ger 坏死性凋亡和细胞焦亡。在散发性 ATAAD 小鼠模型中，主动脉挑战具有高脂肪饮食和血管紧张素 II 输注诱导 EC 中的 RIP3 和 GSDMD，导致内皮细胞通透性过高，纳米颗粒渗透、层间空间扩张、弹性纤维分层和显微切割。 EC-Rip3- /- 或 EC-Gsdmd-/- 显示纳米颗粒浸润和主动脉变性显着减少。] 进一步研究- scATAC-seq 的结果表明，主动脉挑战增加了启动子/增强子的染色质可及性促炎症/促死亡基因，可能是通过激活促炎症转录因子（TF），例如作为IRF3。在人类 EC 中，胞质 DNA 和 STING-TBK1 信号传导是激活该活性的有效上游触发因素。 IRF3 和 BRG1 的变化以及 GSDMD 表达的诱导。该应用程序的目的是测试中心假设是主动脉应激激活 EC 内的炎症信号，从而触发表观遗传促炎症和促死亡程序的诱导，导致 EC 坏死性凋亡/细胞焦亡、屏障功能障碍化、血液成分浸润、主动脉壁变性和 ATAAD 形成。 [目标 1 将检验假设认为RIP3介导的EC坏死性凋亡和GSDMD介导的EC焦亡损害EC屏障功能，从而促进炎性血液成分的浸润并促进主动脉变性 ATAD 形成。在我们的散发性 ATAAD 模型中，我们将比较主动脉变性和 ATAAD 形成在对照小鼠、EC-Rip3-/- 小鼠、EC-Gsdmd-/- 小鼠和用坏死性凋亡/细胞焦亡抑制剂治疗的 WT 小鼠中 tor.] 目标 2 将检验主动脉应激激活炎症信号传导（例如 STING 通路）的假设 EC 内触发促炎症和促死亡程序的表观遗传诱导。我们将表演对未受攻击和受攻击的 WT 小鼠和 EC- 的主动脉中的 EC 进行 scRNA-seq 和 scATAC-seq 分析特定的 Sting KO 小鼠。目标 3 将检验压力信号（例如胞质 DNA）触发传感的假设直接激活染色质重塑复合物（例如， BRG1/SWI/SNF）和 TF（例如 IRF3）可诱导 GSDMD 表达并促进 EC 焦亡。我们将测试培养的人类 EC 中的假设（目标 3A）并[确认人类 ATAAD 主动脉中的机制联系（目标 3B）]。我们期望所提出的研究将为分子机制提供新的见解散发性主动脉变性和 ATAAD 形成，这反过来将促进新疗法的开发用于预防疾病进展及其致命的后遗症。 1