MIcrophysiological systems to model vascular malformations

模拟血管畸形的微生理系统

• 批准号: 10178473
• 负责人: CHRISTOPHER C. W. HUGHES
• 金额: $ 17.92万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-01 至 2021-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10178473
• 关键词: 2019-nCoV; Address; Angiotensin II; Antibodies; Award; Binding Proteins; Blood; Blood - brain barrier anatomy; Blood Vessels; Brain; COVID-19; Cell Surface Receptors; Cell surface; Cells; Cerebral Ischemia; Chronic; Coagulation Process; Complex; Coronavirus; Depressed mood; Disease; Down-Regulation; Drug Targeting; Embolism; Endothelial Cell Inhibitor; Endothelial Cells; Enzymes; Event; Family; Funding; Genes; Heart; Hepatocyte; Human; Immunoglobulin G; Infection; Infection prevention; Inflammation; Inflammatory; Interleukin-6; Kidney; Lead; Liver; Lung; Macrophage Activation; Mediating; Middle East Respiratory Syndrome Coronavirus; Modeling; Mucocutaneous Lymph Node Syndrome; Organ; Parents; Partner in relationship; Patients; Peptide Hydrolases; Peptidyl-Dipeptidase A; Peripheral; Physiological; Pneumonia; Positioning Attribute; Primary Infection; Receptor Cell; Regulation; Renin-Angiotensin System; Reporting; Respiratory physiology; SARS coronavirus; Secondary to; Serine; Serum; Severe Acute Respiratory Syndrome; Site; Smooth Muscle Myocytes; Somatotype; Surface; Technology; Testing; Threonine; Thrombomodulin; Thromboplastin; Thrombosis; Tissue Microarray; Tissues; Up-Regulation; Vascular Endothelial Cell; Vasoconstrictor Agents; Venous; Viral; Viral Proteins; Virus; Virus Diseases; Virus Receptors; cell type; cytokine; cytokine release syndrome; gastrointestinal epithelium; human coronavirus; knock-down; liver function; macrophage; malformation; member; microphysiology system; monocyte; novel; novel coronavirus; pandemic disease; pediatric patients; receptor; renin hypertension; response; secondary infection; side effect; small molecule inhibitor; stem; therapeutic target; tool; vascular inflammation; vasoconstriction

PROJECT SUMMARY COVID19 is caused by SARS-CoV-2, a novel member of the human coronavirus family that includes the closely related SARS-CoV (SARS) and MERS-CoV (MERS) viruses. SARS-CoV-2 viral spike protein binds to human angiotensin converting enzyme-2 (ACE2) on the surface of cells and is then primed by the serine/threonine protease TMPRS22, whereupon the entire complex is internalized by the target cell. ACE2 is expressed by multiple cell types of the body, including lung and gut epithelium (likely the primary sites of initial infection) and vascular endothelial cells (EC) of multiple organs. In addition to the well-described pneumonia-like disease characterized by compromised lung function with subsequent depressed pO2 levels in blood, patients often also show signs of multi-organ involvement, which can include gut, kidney, liver, heart, and brain. Most recently, numerous pediatric patients have been showing signs of Kawasaki disease, a systemic vascular inflammation. Over 30% of COVID19 ICU patients also show signs of thrombosis and 25% suffer venous embolism. Cerebral ischemia, likely due to clot formation, has also been reported. What is not clear is whether this multi-organ involvement is due to secondary infection of these tissues or whether these are all a consequence of systemic hyperinflammation. The sequence of events that could drive systemic hyperinflammation stems from the SARS- CoV-2 mechanism of infection. Angiotensin II (AngII) is an important vasoconstrictor and under normal physiological conditions its level is closely controlled through rapid degradation by ACE2, however, SARS-CoV- 2 entry into cells clears ACE2 from the cell surface, potentially prolonging the action of AngII. Primary consequences of this would be two-fold: prolonged vasoconstriction in the lung (exacerbating poor oxygenation of the blood by the already compromised lungs) and a shift toward a pro-inflammatory state, as it is well established that AngII can drive local vascular inflammation, in large part through the induction of IL-6 in EC and smooth muscle cells (SMC). IL-6 is one of the major drivers of systemic hyperinflammation, and in its most severe form, a so-called “cytokine storm”. IL-6 is also strongly correlated with thrombosis, likely through upregulation of tissue factor on EC and macrophages, and by downregulation of thrombomodulin on EC. Through the parent award we have generated Vascularized Micro-Organs (VMOs), comprised of perfused human vasculature and a surrounding stroma, and have further developed these into Vascularized Micro-Brains (VMBs) incorporating a Blood-Brain Barrier, and Vascularized Micro-Livers (VMLs). Additional cells, including macrophages and SMC have also been incorporated into the VMO. Using these platforms we will address three hypotheses: 1) That the VMO can be used to assess the ability of convalescent serum, soluble ACE2 or small molecule inhibitors to block entry of a SARS-CoV-2 pseudotyped virus; 2) That sustained AngII expression can trigger a hyperinflammatory response from EC and macrophages; and 3) That hyperinflammation can compromise peripheral and BBB vasculature as well as liver function.

项目摘要 COVID19是由SARS-COV-2引起的，Sars-Cov-2是人类冠状病毒家族的新成员 相关的SARS-COV（SARS）和MERS-COV（MERS）病毒。 SARS-COV-2病毒峰值蛋白与人结合 血管紧张素在细胞表面转化酶-2（ACE2），然后通过序列/苏氨酸启动 蛋白酶TMPRS22，因此整个复合物由目标细胞内化。 ACE2由 身体的多种细胞类型，包括肺和肠道上皮（可能是初始感染的主要部位）和 多个器官的血管内皮细胞（EC）。除了描述的肺炎样疾病外 以肺功能受损为特征，随后血液中的PO2降低，患者通常也 显示多器官参与的迹象，其中可能包括肠道，肾脏，肝脏，心脏和大脑。最近， 许多儿科患者一直在显示川崎病的迹象，川崎疾病是一种全身性血管炎症。 超过30％的ICU患者中，超过30％的ICU患者也表现出血栓形成的迹象，25％的患者患有静脉栓塞。大脑 还已经报道了缺血，可能是由于凝块形成引起的。不清楚的是这个多器官是否 参与是由于这些时机的继发感染或这些时间是否是全身性的结果 高炎症。可以从SARS- COV-2感染机制。血管紧张素II（Angii）是重要的血管收缩剂，在正常状态下 生理状况，其水平通过ACE2的快速降解而受到密切控制，但是，SARS-COV- 2进入细胞从细胞表面清除ACE2，可能延长Angii的作用。基本的 这将是两个方面的后果：肺部长时间的血管收缩（加剧氧合不良） 肺已经受损的血液）和向促炎状态的转变，因为它很好 确定ANGII可以通过EC和 平滑肌细胞（SMC）。 IL-6是全身性高炎症的主要驱动因素之一，在其最多 严重的形式，一种所谓的“细胞因子风暴”。 IL-6也与血栓形成密切相关，可能通过 在EC和巨噬细胞上的组织因子上调，以及通过EC上的血栓统治下调。 通过父母奖，我们产生了由灌注的血管化微孔（VMO）（VMO） 人血管和周围的基质，并将其进一步发展为血管化微脑 （VMB）掺入了血脑屏障和血管化微肝（VML）。其他细胞，包括 巨噬细胞和SMC也已纳入VMO。使用这些平台，我们将解决三个 假设：1）VMO可用于评估康复血清，固体ACE2或小的能力 分子抑制剂阻断SARS-COV-2伪型病毒的进入； 2）持续的Angii表达可以 触发来自EC和巨噬细胞的高炎反应； 3）超炎症可以 妥协的外围和BBB脉管系统以及肝功能。