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: 10661665
• 负责人: DENNIS R. VOELKER
• 金额: $ 24.83万
• 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/10661665
• 关键词: Acute; Adult; Affinity; Air; Alleles; Animals; Antiviral Agents; Asthma; Attenuated; Binding; Binding Proteins; Cadherins; Cell Surface Receptors; Cell surface; Cells; Chronic Obstructive Pulmonary Disease; Chronic lung disease; Clinical; Clinical Research; Collaborations; Collection; Complex; Data; Data Set; Economic Burden; Epithelial Cells; Evaluation; Gene Proteins; 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; 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 Cystic Fibrosis; 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; 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) 在导致哮喘恶化方面发挥着重要作用，从而导致疾病和经济恶化 负担。在 RV 中，RV-C 亚组由于其毒力而问题尤其严重。这个提议 专注于 RV-C，并利用源自肺表面活性剂脂质的抗病毒药物改善其效果 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），对抗呼吸道病毒感染和 通过充当诱饵配体来抑制由 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 也可以部分通过直接作用来破坏 RV-C 感染。 与病毒体的相互作用。在本提案中，我们计划研究表面活性剂脂质的机制 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 及其结构如何 类似物使用气液界面 (ALI) 培养物干扰 RV-C 感染/复制。我们当前的数据 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 RNAseq 数据表明 PI 干扰纤毛生成，使宿主细胞难以被纤毛形成。 已感染;因为感染需要成熟、活动的纤毛。相同的 RNAseq 数据集也表明 POPG 的作用 位于与 PI 不同的细胞内位点。目标 1 中还提出了转基因小鼠研究，并将调查 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 使用气液界面离体培养物破坏 RV-C 感染/复制。我们还将进行RNAseq 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 同工型来 确定离体和体内不同 SP-A 结构变体的排序效力。离体 SP- 一项研究将扩展到转基因小鼠的研究，以了解 SP-A 在 完整的肺和整个动物。从本提案中描述的实验中，我们预计会了解到 表面活性剂脂质和蛋白质作为针对加重哮喘的 RV 的抗病毒药物的作用机制 并导致病情加重。我们预计这些信息将确定用于控制 RV 感染的新型试剂 人体中，并产生新的数据，将改善表面活性剂成分的作用。