Vascular, Cardiac, and Lung Alveolar Human Microphysiological Systems for SARS-CoV-2 Drug Screening

用于 SARS-CoV-2 药物筛选的血管、心脏和肺泡人体微生理系统

• 批准号: 10166020
• 负责人: George A Truskey
• 金额: $ 28.06万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-01 至 2022-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10166020
• 关键词: 2019-nCoV; AGTR2 gene; Address; Affect; Alveolar; Antibodies; Appearance; Binding; Binding Proteins; Binding Sites; Biological Assay; Blood Vessels; COVID-19; Captopril; Cardiac; Cardiac Myocytes; Cardiovascular system; Cathepsins; Cell Line; Cells; Clinical Trials; Disease; Dose; Drug Evaluation; Drug Screening; Endocytosis; Endothelial Cells; Epithelial Cells; Exocytosis; Goals; Health; Human; Individual; Infection; International; Lip structure; Lisinopril; Lung; Mesylates; Modeling; Patient Care; Patients; Peptide Hydrolases; Peptidyl-Dipeptidase A; Pharmaceutical Preparations; Process; Proteins; RNA; Recombinants; Resources; Serine Protease; Severe Acute Respiratory Syndrome; Severity of illness; Smooth Muscle Myocytes; Specificity; System; TMPRSS2 gene; Testing; Tissues; Vaccines; Vascular Endothelium; Viral; Viral Proteins; Virus; alternative treatment; alveolar epithelium; base; blocking factor; cell type; design; drug candidate; drug testing; effective therapy; experience; human tissue; in vivo; induced pluripotent stem cell; insight; microphysiology system; overexpression; prevent; receptor; reduce symptoms; response; screening; vaccine effectiveness

Abstract The appearance of SARS-CoV-2 in early 2020 has spurred efforts to limit the disease spread and develop effective treatments. The most promising long-term approach is a vaccine. While some vaccines are entering accelerated clinical trials, it may take 12 or more months before an effective vaccine is available. Even if successful, it may not be possible to treat everyone with a vaccine or the effectiveness of the vaccine may be limited. Given the severity of the disease among a number of those patients, alternative approaches to limit infection should be developed. The goal of this proposal is to use human cardiac, vascular, and lung alveolar microphysiological systems (MPS) to identify possible compounds that block SARS-CoV-2 entry into cells and tissues. While cell binding assays can be used to screen drug candidates, human MPS offer the advantage of testing promising drug candidates under conditions encountered in the body. We propose a tiered approach in which primary cells and cells overexpressing angiotensin converting enzyme (ACE2) are used to identify promising candidates that block SARS-CoV-2 virus entry into cells, and vascular, cardiac, and lung alveolar MPS are used to provide a robust evaluation of drugs that block SARS-CoV-2 binding. The first tier with individual cell types enables a rapid screen and the screen with the microphysiological systems enables testing of the most promising candidates with the tissues most likely to be infected. We will develop the screening assays using a pseudovirus with the SARS-CoV-2 spike protein. In Aim 1, we will develop an assay for pseudovirus binding to ACE2 expressing cells by verifying binding and fusion to cells that express ACE2. We will whether the binding specifically involves the spike protein and determine the levels of binding sites on the cell types used in subsequent aims. In Aim 2, we will screen individual cells types for molecules that block entry into the cell of pseudovirus expressing the spike proteins. Potential drug candidates include those that potentially block spike protein binding (e.g. spike proteins, Captopril, Lisinopril, human recombinant soluble ACE2, and antibodies to the spike protein or ACE2) and those inhibiting Transmembrane Serine Protease 2 (TMPRSS2), activity (e.g. camostat mesylate, nafamostat mesylate). In Aim 3, we will test most promising compounds in vascular, cardiac and lung microphysiological systems and compare against results from 2D studies. We will also examine the relationship between drug blocking and factors that affect ACE2 expression.

抽象的 SARS-COV-2在2020年初的出现刺激了限制疾病传播和发展的努力 有效的治疗方法。最有希望的长期方法是一种疫苗。当一些疫苗进入 加速的临床试验，可能需要12个月或更长时间才能获得有效的疫苗。即使 成功，可能不可能用疫苗治疗每个人或疫苗的有效性可能是 有限的。鉴于许多患者中疾病的严重程度，限制的替代方法 应该开发感染。该建议的目的是使用人类心脏，血管和肺肺泡 微生物生理系统（MPS）识别可能阻止SARS-COV-2进入细胞和的化合物 组织。虽然细胞结合测定可用于筛选候选药物，但人MP提供了优势 在体内遇到的条件下测试有希望的候选药物。我们提出了一种分层的方法 哪些过表达血管紧张素转化酶（ACE2）过表达的细胞和细胞用于鉴定 有望阻止SARS-COV-2病毒进入细胞的候选人，血管，心脏和肺肺泡 MP用于对阻断SARS-COV-2结合的药物进行强有力的评估。与 单个细胞类型可以快速屏幕和带有微生物生理系统的屏幕实现测试 最有前途的候选人最有可能被感染的组织。我们将开发筛选 使用伪病毒与SARS-COV-2尖峰蛋白进行测定。在AIM 1中，我们将开发一个用于 假病毒与ACE2结合通过验证表达ACE2的细胞的结合和融合来表达细胞。我们 结合是否特异性涉及尖峰蛋白并确定结合位点的水平 后续目标中使用的细胞类型。在AIM 2中，我们将筛选单个细胞类型的分子类型 进入表达尖峰蛋白的假病毒细胞的进入。潜在的候选毒品包括那些 潜在地阻断尖峰蛋白结合（例如，尖峰蛋白，卡托普利，丽索普利，人类重组可溶性 ACE2，以及对尖峰蛋白或ACE2的抗体以及抑制跨膜丝氨酸蛋白酶2的抗体 （TMPRSS2），活性（例如，麦甲酸酯，脂肪酯麦甲酸酯）。在AIM 3中，我们将测试最有前途的 血管，心脏和肺微生物生理系统中的化合物与2D的结果进行比较 研究。我们还将检查药物阻断与影响ACE2表达的因素之间的关系。