Novel Role of Thrombospondin-1 in Protection against Rupture of Abdominal Aortic Aneurysm

Thrombospondin-1 在预防腹主动脉瘤破裂中的新作用

基本信息

批准号：
10609876
负责人：
Bo Liu
金额：
$ 45.99万
依托单位：
UNIVERSITY OF WISCONSIN-MADISON
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-05-01 至 2025-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10609876
关键词：
Abdominal Aortic Aneurysm Actins Address Adoptive Transfer Affect Aneurysm Angiotensin II Animals Aorta Aortic Aneurysm Aortic Rupture Apoptotic Area Atherosclerosis Blood Vessels Bone Marrow Breeding CD47 gene Cells Clinical Complication Coupled Data Deoxyribonuclease I Detection Development Dilatation - action Elastases Female Gene Deletion Growth Human Imipramine Immunohistochemistry Impairment In Situ Hybridization Incidence Infiltration Inflammation Infusion procedures Knock-out Knockout Mice Knowledge Life Style Low-Density Lipoproteins LoxP-flanked allele Macrophage Mediating Modeling Molecular Molecular Profiling Monitor Mouse Strains Mus Operative Surgical Procedures Patients Phagocytosis Phenotype Play Polymers Preventive Proteins Publications Reporting Research Role Rupture Ruptured Abdominal Aortic Aneurysm Ruptured Aneurysm Small Interfering RNA Source System THBS1 gene Telemetry Testing Thrombospondin 1 Tissues United States Vascular Diseases atherosclerotic plaque rupture beta Aminopropionitrile conditional knockout extracellular human disease hypercholesterolemia induced pluripotent stem cell inhibitor knock-down male migration mortality mouse model neutrophil novel polymerization receptor response single-cell RNA sequencing spatial relationship therapeutic development ultrasound uptake

项目摘要

Abdominal aortic aneurysm (AAA) is the progressive weakening and dilation of the aorta. A substantial knowledge gap exists in the understanding of molecular mechanisms responsible for aneurysm rupture, the major cause of mortality among AAA patients. Following our prior report of elevated thrombospondin-1 (TSP1) in human and mouse aneurysmal tissues, we conducted single-cell RNA sequencing (scRNA-seq) analysis and identified macrophages (Mɸs) being the primary source of elevated TSP1 in mouse aneurysmal aorta. We subsequently generated Mɸ-specific Thbs1 knockout mice (Thbs1∆Mɸ) by crossing Lyz2-Cre with our newly constructed Thbs1flox/flox mice. When subjected to aneurysm induction by angiotensin II (Ang II) coupled with hypercholesterolemia, over 60% of Thbs1∆Mɸ died due to AAA rupture, an incidence that was 2.6 times higher than Thbs1wt. Intriguingly, Thbs1∆Mɸ mice that survived to the end of 28-day Ang II infusion showed less aneurysm dilation than Thbs1wt. Smaller aneurysmal expansion was also found when Thbs1∆Mɸ mice were challenged with perivascular application of CaCl2, an AAA model that does not produce rupture. We propose two specific aims to delineate the mechanisms through which Mɸ-specific Thbs1 gene deletion differentially affects aortic dilation and rupture with an emphasis on AAA rupture. Specific Aim 1 devotes to establishing the rupture- preventive function of Mɸ TSP1. Specifically, we will determine the aortic responses proceeding lethal rupture in male and female Thbs1∆Mɸ mice in the Ang II model followed by identifying rupture-associated molecular signatures through scRNA-seq, in situ hybridization and immunostaining. Furthermore, we will examine the effects of Mɸ-specific Thbs1 knockout using a different murine model that produces rupture in advanced stages of aneurysm, which is more relevant to human AAA than the early rupture produced by the Ang II model. Specific Aim 2 focuses on investigating molecular mechanisms of aneurysm rupture. Preliminary studies showed that compared to wildtype, Thbs1-/- Mɸs had significantly reduced ability to migrate or to engulf apoptotic cells as well as neutrophil extracellular traps (NETs). We will test whether Mɸ TSP1 promotes NET clearance through CD47- mediated actin polymerization. Next, we will establish the causal effect of impaired Mɸ migration and phagocytosis on aneurysm rupture of Thbs1∆Mɸ mice. We will first determine whether NET burden is increased in Thbs1∆Mɸ died from rupture, and the spatial relationship between NETs and Mɸs. Second, we will test whether restoring Mɸ migration in Thbs1∆Mɸ reduces NET accumulation via adoptive transfer strategies. Furthermore, we will examine whether enhancing or attenuating NET clearance affect aneurysm rupture in Thbs1∆Mɸ. Lastly, we will analyze TSP1 expression and its association with Mɸ and NET accumulation in ruptured and non-ruptured human AAA tissues. By dissecting the multifaceted functions of Mɸs through TSP1 manipulations, this project will produce significant impact on the understanding of aneurysm rupture.

腹主动脉瘤（AAA）是主动脉进行性减弱和扩张。对导致动脉瘤破裂的分子机制的理解存在知识差距根据我们之前的报告，血小板反应蛋白-1 (TSP1) 升高。在人类和小鼠动脉瘤组织中，我们进行了单细胞 RNA 测序 (scRNA-seq) 分析并确定巨噬细胞 (Mɸs) 是小鼠主动脉瘤 TSP1 升高的主要来源。随后通过将 Lyz2-Cre 与我们新的杂交产生了 Mɸ 特异性 Thbs1 敲除小鼠 (Thbs1ΔMɸ) 构建了 Thbs1flox/flox 小鼠，当接受血管紧张素 II (Ang II) 联合诱导动脉瘤时。高胆固醇血症，超过 60% 的 Thbs1ΔMɸ 因 AAA 破裂而死亡，发病率高出 2.6 倍有趣的是，存活至 28 天 Ang II 输注结束的 Thbs1ΔMɸ 小鼠显示出较少的动脉瘤。当 Thbs1ΔMɸ 小鼠受到挑战时，也发现了比 Thbs1wt 更小的动脉瘤扩张。 CaCl2 的血管周围应用，一种不会产生破裂的 AAA 模型，我们提出了两个具体目标。描述 Mɸ 特异性 Thbs1 基因缺失对主动脉产生不同影响的机制扩张和破裂，重点是 AAA 破裂。具体目标 1 致力于确定破裂。 Mɸ TSP1 的预防功能具体来说，我们将确定致命性破裂时的主动脉反应。 Ang II 模型中的雄性和雌性 Thbs1ΔMɸ 小鼠，然后识别破裂相关分子此外，我们将通过 scRNA-seq、原位杂交和免疫染色来检查特征。使用不同的小鼠模型进行 Mɸ 特异性 Thbs1 敲除的效果，该模型在晚期阶段产生破裂动脉瘤，这比 Ang II 模型产生的早期破裂与人类 AAA 更相关。目标 2 重点研究动脉瘤破裂的分子机制。初步研究表明。与野生型相比，Thbs1-/- Mɸs 迁移或吞噬凋亡细胞的能力也显着降低作为中性粒细胞胞外陷阱 (NET)，我们将测试 Mɸ TSP1 是否通过 CD47- 促进 NET 清除。接下来，我们将确定 Mɸ 迁移受损的因果效应和 Thbs1ΔMɸ小鼠动脉瘤破裂的吞噬作用我们首先确定NET负荷是否增加。在 Thbs1ΔMɸ 中，NETs 和 Mɸs 之间的空间关系是否因破裂而死亡。恢复 Thbs1ΔMɸ 中的 Mɸ 迁移可通过采用转移策略减少 NET 积累。将检查增强或减弱 NET 清除是否会影响 Thbs1ΔMɸ 的动脉瘤破裂。将分析破裂和非破裂中 TSP1 表达及其与 Mɸ 和 NET 积累的关联该项目通过 TSP1 操作剖析 Mɸs 的多方面功能。将对动脉瘤破裂的认识产生重大影响。

项目成果

期刊论文数量（1）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Bim Expression Modulates Branching Morphogenesis of the Epithelium and Endothelium.

DOI：
10.3390/biom12091295
发表时间：
2022-09-14
期刊：
BIOMOLECULES
影响因子：
5.5
作者：
Sorenson, Christine M.;Song, Yong-Seok;Wang, Shoujian;Darjatmoko, Soesiawati R.;Saghiri, Mohammad Ali;Ranji, Mahsa;Sheibani, Nader
通讯作者：
Sheibani, Nader