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.

摘要 2020年初SARS-CoV-2的出现促使人们努力限制疾病的传播和发展 有效的治疗。最有希望的长期方法是疫苗。当一些疫苗进入 加速临床试验，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，它可能不可能用疫苗治疗每个人，或者疫苗的有效性可能会降低。 有限公司考虑到这些患者中的一些疾病的严重程度，限制 感染应该发展。这项提案的目标是利用人体心脏、血管和肺泡 微生理系统（MPS），以确定可能的化合物，阻止SARS-CoV-2进入细胞， 组织中虽然细胞结合测定可用于筛选候选药物，但人MPS提供了以下优点： 在体内遇到的条件下测试有希望的候选药物。我们提出了一个分层的方法， 使用哪种原代细胞和过表达血管紧张素转化酶（ACE 2）的细胞来鉴定 有希望的候选人，阻止SARS-CoV-2病毒进入细胞，血管，心脏和肺泡， MPS用于提供阻断SARS-CoV-2结合的药物的稳健评价。第一层， 单个细胞类型能够进行快速筛选，并且使用微生理系统进行筛选能够进行测试 最有希望的候选人的组织最有可能被感染。我们将发展筛选 使用具有SARS-CoV-2刺突蛋白的假病毒进行测定。在目标1中，我们将开发一种测定方法， 通过验证与表达ACE 2的细胞的结合和融合，将假病毒与表达ACE 2的细胞结合。我们 将确定结合是否特异性地涉及刺突蛋白，并确定刺突蛋白上结合位点的水平。 用于后续目标的细胞类型。在目标2中，我们将筛选单个细胞类型的分子， 表达刺突蛋白的假病毒进入细胞。潜在的候选药物包括那些 潜在地阻断刺突蛋白结合（例如刺突蛋白、卡托普利、赖诺普利、人重组可溶性 ACE 2，以及刺突蛋白或ACE 2的抗体）和抑制跨膜丝氨酸蛋白酶2的那些 （TMPRSS 2）、活性（例如卡莫司他甲磺酸盐、萘莫司他甲磺酸盐）。在目标3中，我们将测试最有希望的 化合物在血管，心脏和肺微生理系统中的作用，并与2D结果进行比较 问题研究我们还将研究药物阻断和影响ACE 2表达的因素之间的关系。