Surfactant Lipid and Protein Inhibition of Rhinovirus Infections

表面活性剂脂质和蛋白质对鼻病毒感染的抑制作用

• 批准号: 10261955
• 负责人: DENNIS R. VOELKER
• 金额: $ 33.26万
• 依托单位: UNIVERSITY OF ARIZONA
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-06-01 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10261955
• 关键词: Acute; Adult; Affinity; Air; Alleles; Animals; Antiviral Agents; Asthma; Attenuated; Binding; Cadherins; Cell Surface Receptors; Cell surface; Cells; Chronic Obstructive Airway Disease; Chronic lung disease; Clinical; Clinical Research; Collaborations; Collection; Complex; Cystic Fibrosis; Data; Data Set; Economic Burden; Epithelial Cells; Evaluation; Genes; Genetic; Genetic Polymorphism; Genetic Transcription; Homologous Gene; Hospitalization; Human; Immunization; Infection; Inflammation; Inflammatory; Inflammatory Response Pathway; Influenza A virus; Innate Immune System; Interferons; Interstitial Lung Diseases; Irrigation; Lead; Learning; Length; Letters; Ligands; Lipids; Liquid substance; Lung; Lung diseases; Mediating; Minor; Mus; Nasal Epithelium; Natural Immunity; Outcome; Participant; Patients; Phosphatidylglycerols; Phosphatidylinositols; Phospholipids; Play; Prevention; Process; Production; Prophylactic treatment; Protein Inhibition; Protein Isoforms; Proteins; Pulmonary Surfactant-Associated Protein A; Pulmonary Surfactants; Pyroglyphidae; Reagent; Receptor Activation; Refractory; Replication-Associated Process; Respiratory syncytial virus; Rhinovirus; Rhinovirus infection; Role; Serotyping; Serum; Site; Structure; Subgroup; System; Therapeutic; Toll-like receptors; Transcription Process; Transgenic Mice; Variant; Viral; Viral Load result; Viral Physiology; Viral Respiratory Tract Infection; Virion; Virulence; Virulent; Virus; Virus Diseases; Virus Replication; Wheezing; Whole Organism; Work; analog; anti-viral efficacy; asthma exacerbation; asthmatic; asthmatic patient; burden of illness; cilium motility; early childhood; experimental study; genetic variant; human coronavirus; improved; in vivo; mouse model; novel; patient population; prevent; programs; replication factor C; response; surfactant; targeted agent; targeted treatment; transcriptome; transcriptome sequencing; viral entry inhibitor

Rhinoviruses (RVs) play a major role in causing asthma exacerbations, which drive disease and economic burdens. Among the RVs the RV-C subgroup is especially problematic, due to its virulence. This proposal is focused on RV-C, and ameliorating its effects with anti-viral agents derived from pulmonary surfactant lipids and proteins. Two minor anionic phospholipids of pulmonary surfactant, palmitoyl-oleoyl- phosphatidylglycerol (POPG) and phosphatidylinositol (PI), antagonize respiratory viral infections and suppress inflammatory sequelae triggered by Toll-like receptor activation, by acting as decoy ligands. In addition, the pulmonary surfactant protein, SP-A, also disrupts RV-C infections, in part by direct interactions with virions. In this proposal we plan to investigate the mechanisms by which surfactant lipids and proteins inhibit RV infections and replication. In Aim 1, we will examine how POPG and PI and their structural analogs interfere with RV-C infection/replication using Air-Liquid Interface (ALI) cultures. Our current data support a mechanism in which the lipids act intracellularly to alter the infection/replication processes. Indeed, preliminary RNAseq data suggest that PI interferes with ciliagenesis, rendering host cells refractory to being infected; since infection requires mature, motile cilia. The same RNAseq data set also suggests that POPG acts at a different intracellular site from PI. Transgenic mouse studies are also proposed in Aim 1 and will investigate the efficacy of the anti-viral phospholipids in the context of the whole organism. In Aim 2, we will investigate how SP-A disrupts RV-C infection/replication using Air-Liquid-interface cultures ex vivo. We will also perform RNAseq analysis to probe the host cell and viral transcriptomes. We will use our collection of purified SP-A isoforms to determine the rank order potency of different SP-A structural variants both ex vivo and in vivo. The ex vivo SP- A studies will be expanded to studies in transgenic mice to understand the actions of SP-A in the context of an intact lung and whole animal. From the experiments described in this proposal we anticipate learning the mechanisms of action of surfactant lipids and proteins as anti-viral agents targeting RVs that aggravate asthma and cause exacerbations. We expect this information will identify novel reagents for controlling RV infections in humans, and lead to new data that will improve the actions of the surfactant constituents.

鼻病毒（RV）在引起哮喘急性发作方面发挥着重要作用，这导致疾病和经济损失。 负担在RV中，RV-C亚组由于其毒力而特别成问题。这项建议 目前的研究重点是RV-C，并使用来自肺表面活性物质脂质的抗病毒药物改善其作用 和蛋白质。肺表面活性剂的两种次要阴离子磷脂，棕榈酰-油酰- 磷脂酰甘油（POPG）和磷脂酰肌醇（PI），拮抗呼吸道病毒感染， 通过作为诱饵配体，抑制由Toll样受体激活引发的炎症后遗症。 此外，肺表面活性蛋白SP-A也可部分通过直接抑制RV-C感染而破坏RV-C感染。 与病毒粒子的相互作用。在这个提议中，我们计划研究表面活性剂脂质 和蛋白质抑制RV感染和复制。在目标1中，我们将研究POPG和PI及其结构 类似物干扰使用气液界面（ALI）培养的RV-C感染/复制。我们目前的数据 支持脂质在细胞内作用以改变感染/复制过程的机制。的确， 初步的RNAseq数据表明，PI干扰纤毛发生，使宿主细胞难以被 感染;因为感染需要成熟的、能动的纤毛。同样的RNAseq数据集也表明，POPG的行为 位于与PI不同的细胞内位点。目标1中也提出了转基因小鼠研究， 抗病毒磷脂在整个生物体中的功效。在目标2中，我们将研究如何 SP-A使用离体空气-液体界面培养物破坏RV-C感染/复制。我们还将进行RNA测序 分析以探测宿主细胞和病毒转录组。我们将使用我们收集的纯化SP-A同种型， 测定不同SP-A结构变体在体外和体内的等级效力。体外SP- A研究将扩展到转基因小鼠的研究，以了解SP-A在一种环境中的作用。 完整肺和整个动物。从本提案中描述的实验中，我们预计将学习 表面活性剂脂质和蛋白质作为靶向加重哮喘的RV的抗病毒药物的作用机制 并导致病情加重我们希望这些信息将确定控制RV感染的新试剂 在人类中，并导致新的数据，将改善表面活性剂成分的行动。