VPS34 inhibitors as SARS-CoV-2 antivirals

VPS34 抑制剂作为 SARS-CoV-2 抗病毒药物

• 批准号: 10238577
• 负责人: Christopher F Basler
• 金额: $ 1.31万
• 依托单位: GEORGIA STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-01 至 2021-11-15
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10238577
• 关键词: 1-Phosphatidylinositol 3-Kinase; 2019-nCoV; Acyl Coenzyme A; Address; Antiviral Agents; Autophagocytosis; Biological Assay; COVID-19; COVID-19 patient; COVID-19 treatment; Cell membrane; Cells; Clinical; Coenzyme A Ligases; Coronavirus; Coronavirus Infections; Data; Development; Diabetes Mellitus; Endoplasmic Reticulum Degradation Pathway; Enzyme Inhibitor Drugs; Enzymes; Exhibits; FDA approved; Fatty-acid synthase; Generations; Genetic; Growth; Hamsters; Health; Human; Impairment; Infection; Integration Host Factors; Knock-out; Lipids; Malignant Neoplasms; Measures; Medical; Membrane; Membrane Biology; Mesocricetus auratus; Metabolic; Microscopy; Modeling; Natural Immunity; Nonstructural Protein; Obesity; Oral; Organelles; Outcome; Palmitates; Pathway interactions; Pharmacology; Phosphotransferases; Play; Proteins; Proteomics; RNA Viruses; RNA chemical synthesis; Regulation; Replicon; Role; SARS-CoV-2 infection; Site; Societies; Testing; Therapeutic; Therapeutic Intervention; Time; Vesicle; Viral; Virus; Virus Replication; base; betacoronavirus; cytotoxicity; drug repurposing; in vivo; inhibitor/antagonist; innovation; insight; interest; knock-down; lipid metabolism; lipid transport; long chain fatty acid; member; novel; orlistat; pandemic disease; phosphatidylinositol 3-phosphate; remdesivir; small molecule; small molecule inhibitor; targeted treatment; therapeutic development; therapeutic target; trafficking; transcriptomics; triacsin C; viral RNA

Summary SARS-CoV-2, a Betacoronavirus genus, is an enveloped positive-sense, RNA virus responsible for a current pandemic. Because of its profound impact on society and human health there is an urgent need to understand SARS-CoV-2 replication requirements and to identify therapeutic strategies. Repurposing drugs developed for other purposes may provide a shortcut to therapeutic development. The use of compounds known to target specific host factors may also elucidate key pathways needed for virus replication. Coronavirus (CoV) replication involves multiple critical interactions with host cell membranes. One of the most striking features of CoV infection is the establishment of membrane-associated replication organelles that serve as the main sites of viral RNA synthesis. The origin of these membrane organelles is incompletely understood. Because the specific host pathways required for SARS-CoV-2 replication organelle formation are not defined, we asked whether SARS-CoV-2 is susceptible to modulators of lipid metabolism by assessing the sensitivity of the virus to VPS34 inhibitors of VPS34, a lipid kinase required for autophagy and endosomal trafficking; Triacsin C, an inhibitor of long chain fatty acyl CoA synthetase (ACSL) and Orlistat, an inhibitor of fatty acid synthase (FASN). Our preliminary data indicate that inhibitors of VPS34 potently inhibited SARS-CoV-2 replication, whereas an FDA- approved inhibitor of a different class of PI3K had minimal effect on replication. Targeting FASN and ACSL also impairs SARS-CoV-2 replication. These data suggest that VPS34, ACSL and FASN play important roles in replication center formation and virus growth and suggest these enzymes as therapeutic targets. We will test the hypothesis that VPS34, ACSL and FASN are critical for SARS-Cov-2 infection by evaluating additional small molecule inhibitors of these enzymes and by measuring SARS-CoV-2 replication in genetic knockdowns or knockouts of these host enzymes. We will define mechanisms of inhibition and test the hypothesis that generation of membrane-associated viral replication centers will be disrupted. Finally, we will assess the in vivo efficacy of VSP34 inhibitor PIK-III and systemically administered Orlistat in SARS-coV-2- infected hamsters to evaluate the therapeutic potential of inhibitors of lipid metabolism.

摘要 SARS-CoV-2是一种β冠状病毒属，是一种有包膜的阳性RNA病毒 以应对当前的大流行。由于它对社会和人类健康的深刻影响， 迫切需要了解SARS-CoV-2复制要求并确定治疗方法 战略。为其他目的开发的药物的再利用可能为治疗提供一条捷径 发展。使用已知的针对特定宿主因子的化合物也可以阐明关键 病毒复制所需的途径。冠状病毒(CoV)复制涉及多个关键 与宿主细胞膜的相互作用。冠状病毒感染最显著的特征之一是 作为病毒主要部位的膜相关复制细胞器的建立 RNA合成。这些膜细胞器的起源尚不完全清楚。因为 SARS-CoV-2复制细胞器形成所需的特定宿主途径不是 定义，我们询问SARS-CoV-2是否对脂代谢调节剂易感，通过 评估病毒对Vps34抑制剂的敏感性，Vps34是一种脂酶，需要 自噬和内体运输；Triacsin C，长链脂肪酰辅酶A的抑制剂 脂肪酸合成酶(ACSL)和脂肪酸合成酶(FASN)抑制剂奥利司他。我们的初步数据 表明Vps34的抑制剂有效地抑制了SARS-CoV-2的复制，而FDA- 批准的另一类PI3K的抑制剂对复制的影响很小。目标FASN ACSL还可抑制SARS-CoV-2的复制。这些数据表明，Vps34、ACSL和 FASN在复制中心的形成和病毒的生长中起着重要的作用，并提示这些 以酶为治疗靶点。我们将测试Vps34、ACSL和FASN是 通过评估其他小分子抑制剂对SARS-CoV-2感染的关键作用 并通过测量SARS-CoV-2在这些基因敲除或敲除中的复制 宿主酶。我们将定义抑制机制并检验生成的假设 膜相关病毒复制中心的一部分将被破坏。最后，我们将评估In VSP34抑制剂PIK-III和全身应用奥利司他对SARS-CoV-2的体内疗效 以评估脂代谢抑制剂的治疗潜力。