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; 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补充：基于聚糖的新型治疗对黄病毒的机制和体内活性 内皮渗透性和血管泄漏 评估靶向SARS-COV-2感染的治疗剂和定义的致病机制 SARS-COV-2触发的肺功能障碍 抽象的 新兴的严重急性呼吸系统综合症 - 科罗纳病毒2（SARS-COV-2），一种原因 2019年冠状病毒病（Covid-19）正在迅速蔓延到世界各地，已经影响199个国家， 预计将感染多达60％的人口，迄今为止，病例死亡率约为4.4％。新颖的治疗学 迫切需要，并且由于我们目前正在研究环糊精的抗氟化病毒特性 具有广泛抗病毒活性的化合物（CD），我们在这里提议扩大此研究以测试这些研究 人支气管上皮细胞中抗SARS-COV-2活性的化合物（AIM 1）。首先，我们将快速追踪 在我们的集合中，FDA批准的CD，然后筛选其余的。 OSC Ralph Baric博士（UNC）将确认我们的最多 在原代人细胞中有希望的候选人，并协作建立进一步的研究以在他的身上进行测试 鼠标模型。基于以前的文献和通过父母赠款获取的数据，我们假设 这些化合物可能通过灭活病毒体具有直接的病毒活性，并可能抑制细胞 通过阻断SARS-COV-2尖峰糖蛋白（S）与聚糖和/或病毒相互作用来附着 受体血管紧张素在细胞表面转化酶2（ACE2）。尽管感染的患者屈服于 涉及血管泄漏的急性呼吸窘迫综合征（ARDS），该病理的病毒触发因素是 不清楚。使用SARS-COV-1进行的实验发现，ACE2的内在化与病毒颗粒在 感染可降低细胞表面的ACE2水平，从而导致血管紧张素II活性增加。血管紧张素 据信II活性导致血管活性分子（如血管内皮生长因子）上调 F（VEGF）和细胞间连接的破坏，都引起血管泄漏。在AIM 2中，我们的优化系统 用于研究内皮细胞功能障碍的研究，这是由于使用黄病毒多年而引起的 家长赠款将用于研究病毒S蛋白诱导的SARS-COV-2血管病理 和二级调解人等VEGF。正如我们已经观察到的体外抗裂孔和抗病毒特性 对于某些经过测试的CD，我们还建议将这些候选者作为治疗Covid-19疾病的治疗剂进行测试 表现。因此，该补充赠款提案有可能定义SARS-COV-2 S-的触发器 介导的血管泄漏，导致ARDS，以及将CD作为针对病毒感染的治疗剂 直接和间接通过下游发病机理。证明是FDA批准的CD和衍生物 有效，因为199. 199的治疗可能会迅速开发出来在患者中的潜在用途。 1