Administrative Supplement to R21: Mechanism and in vivo activity of novel glycan-based therapy against flavivirus endothelial permeability and vascular leak

R21 的行政补充：针对黄病毒内皮通透性和血管渗漏的新型聚糖疗法的机制和体内活性

• 批准号: 10265787
• 负责人: Eva Harris
• 金额: $ 20.77万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-03-01 至 2023-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10265787
• 关键词: 2019-nCoV; ACE2; Administrative Supplement; Adult Respiratory Distress Syndrome; Affect; Angiotensin II; Antiviral Agents; Applications Grants; Blood Vessels; COVID-19; COVID-19 pandemic; COVID-19 treatment; Case Fatality Rates; Cell surface; Cell-Matrix Junction; Cells; Collection; Country; Cyclodextrins; Data; Disease; Endothelial Cells; Endothelium; Epithelial Cells; Exploratory/Developmental Grant; F Factor; FDA approved; Flavivirus; Functional disorder; Human; In Vitro; Infection; Intercellular Junctions; Investigation; Literature; Lung; Mediating; Mediator of activation protein; Molecular; Pathogenesis; Pathogenicity; Pathology; Patients; Permeability; Polysaccharides; Population; Property; Pulmonary Edema; Rest; SARS coronavirus; SARS-CoV-2 infection; SARS-CoV-2 inhibitor; SARS-CoV-2 spike protein; System; Testing; Therapeutic; Up-Regulation; Vaccines; Vascular Endothelial Growth Factors; Vascular Permeabilities; Viral; Viral Proteins; Virion; Virus; Virus Diseases; Virus Receptors; Work; base; bronchial epithelium; experimental study; in vivo; mouse model; novel; novel therapeutics; parent grant; therapeutic evaluation; therapeutic target; virucide

Supplement to R21: Mechanism and in vivo activity of novel glycan-based therapy against flavivirus endothelial permeability and vascular leak Evaluation of therapeutics targeting SARS-CoV-2 infection and defining pathogenic mechanisms of SARS-CoV-2-triggered pulmonary dysfunction Abstract The emerging severe acute respiratory syndrome-corona virus 2 (SARS-CoV-2), the causative agent of coronavirus disease 2019 (COVID-19), is spreading rapidly across the world, already affecting 199 countries, and predicted to infect up to 60% of the population, with a ~4.4% case fatality rate to date. Novel therapeutics are desperately needed, and as we are currently investigating anti-flavivirus properties of cyclodextrin compounds (CDs), which have broad antiviral activity, we propose here to expand this investigation to test these compounds for anti-SARS-CoV-2 activity in human bronchial epithelial cells (Aim 1). First, we will fast-track the FDA-approved CDs in our collection, and then screen the rest. OSC Dr. Ralph Baric (UNC) will confirm our most promising candidates in primary human cells and collaborate on setting up further studies to test them in his mouse models. Based on previous literature and data acquired through the parent grant, we hypothesize that these compounds may have direct virucidal activity by inactivating virions, as well as potentially inhibiting cell attachment by blocking the SARS-CoV-2 spike glycoprotein (S) from interacting with glycans and/or the viral receptor angiotensin converting enzyme 2 (ACE2) on the cell surface. Though infected patients succumb to acute respiratory distress syndrome (ARDS) involving vascular leak, the viral triggers of this pathology are unclear. Experiments with SARS-CoV-1 found that internalization of ACE2 along with virus particles upon infection reduces ACE2 levels on the cell surface, resulting in increased angiotensin II activity. The angiotensin II activity is believed to result in upregulation of vasoactive molecules such as vascular endothelial growth factor F (VEGF) and disruption of intercellular junctions, both inducing vascular leak. In Aim 2, our optimized system for the study of endothelial cell dysfunction, resulting from many years of work with flaviviruses and included in the parent grant, will be applied to investigate SARS-CoV-2 vascular pathology induced by the viral S protein and secondary mediators like VEGF. As we have already observed in vitro anti-leak as well as antiviral properties for some of the tested CDs, we also propose to test these candidates as therapeutics to treat COVID-19 disease manifestations. As such, this supplemental grant proposal has the potential to define triggers of SARS-CoV-2 S- mediated vascular leak, contributing to ARDS, as well as testing CDs as therapeutics targeting viral infection directly and indirectly via downstream pathogenesis. FDA-approved CDs and derivatives that prove to be effective as COVID-19 treatments have the potential to be rapidly developed for potential use in patients. 1

R21的补充：基于聚糖的抗黄病毒治疗的机制和体内活性 内皮通透性和血管渗漏 SARS-CoV-2感染靶向治疗药物的评价及其致病机制的确定 SARS-CoV-2引发的肺功能障碍 摘要 新出现的严重急性呼吸道综合征冠状病毒2型（SARS-CoV-2）， 2019冠状病毒病（COVID-19）正在全球迅速蔓延，已经影响到199个国家， 预计将感染高达60%的人口，迄今为止的病死率约为4.4%。新疗法 我们目前正在研究环糊精的抗黄病毒特性， 化合物（CD），具有广泛的抗病毒活性，我们建议在这里扩大这项调查，以测试这些 化合物在人支气管上皮细胞中的抗SARS-CoV-2活性（Aim 1）。首先，我们将快速跟踪 FDA批准的CD，然后筛选剩下的。OSC博士拉尔夫·巴里克（Ralph Baric）将确认我们最 有希望的候选人在原代人类细胞，并合作建立进一步的研究，以测试他们在他的 小鼠模型。根据以前的文献和通过父母资助获得的数据，我们假设， 这些化合物可通过灭活病毒体以及潜在地抑制细胞增殖而具有直接杀病毒活性。 通过阻断SARS-CoV-2刺突糖蛋白（S）与聚糖和/或病毒结合蛋白相互作用， 受体血管紧张素转换酶2（ACE 2）的细胞表面。虽然感染的病人会死于 急性呼吸窘迫综合征（ARDS）涉及血管渗漏，这种病理的病毒触发因素是 不清楚用SARS-CoV-1进行的实验发现，ACE 2沿着病毒颗粒的内化， 感染降低了细胞表面的ACE 2水平，导致血管紧张素II活性增加。血管紧张素 II活性被认为导致血管活性分子如血管内皮生长因子的上调 F（VEGF）和细胞间连接的破坏，均诱导血管渗漏。在目标2中，我们的优化系统 用于研究内皮细胞功能障碍，这是多年来对黄病毒的研究结果， 该基金将用于研究病毒S蛋白诱导的SARS-CoV-2血管病变 和次要介质如VEGF。正如我们已经观察到的体外抗泄漏以及抗病毒特性 对于一些经过测试的CD，我们还建议测试这些候选药物作为治疗COVID-19疾病的疗法 表现。因此，这一补充拨款提案有可能定义SARS-CoV-2 S的触发因素， 介导的血管渗漏，导致ARDS，以及测试CD作为靶向病毒感染的治疗剂 直接和间接通过下游发病机制。FDA批准的CD和衍生品，证明是 有效，因为COVID-19治疗有可能迅速开发用于患者的潜在用途。 1