Surfactant Lipid and Protein Inhibition of Rhinovirus Infections

鼻病毒(RV)在导致哮喘加重中起主要作用，哮喘加重会导致疾病和经济 负担。在RV中，RV-C亚群尤其有问题，因为它的毒力。这项建议 重点是RV-C，并用肺表面活性物质脂类衍生的抗病毒药物改善其效果 和蛋白质。肺表面活性物质的两种少量阴离子磷脂，棕榈酰-油酰基- 磷脂酰甘油(POPG)和磷脂酰肌醇(PI)，拮抗呼吸道病毒感染和 通过充当诱饵配体，抑制Toll样受体激活引发的炎性后遗症。 此外，肺表面活性物质蛋白SP-A也可部分通过直接抑制RV-C感染 与病毒粒子的相互作用。在这项提案中，我们计划研究表面活性物质脂质的作用机制。 蛋白质抑制轮状病毒感染和复制。在目标1中，我们将研究POPG和PI及其结构 类似物利用气液界面(ALI)培养干扰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干扰纤毛形成，使宿主细胞难以 感染；因为感染需要成熟的、活动的纤毛。同样的RNAseq数据集也表明POPG起作用 在与PI不同的细胞内位置。转基因小鼠的研究也在目标1中提出，并将进行调查 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通过体外空气-液体-界面培养破坏RV-C感染/复制。我们还将执行RNAseq 分析以探测宿主细胞和病毒转录本。我们将使用我们收集的纯化的SP-A亚型来 测定不同SP-A结构变异体在体内和体外的秩序效价。体外SP- A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，我们预计将了解 肺表面活性物质、脂质和蛋白质作为抗病毒药物靶向加重哮喘的RV的作用机制 并导致病情恶化。我们期望这一信息将确定控制轮状病毒感染的新试剂。 在人类身上，并导致新的数据，将改善表面活性成分的作用。

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.