Lung MIWI2 and the host defense against Influenza A virus

肺 MIWI2 和宿主对甲型流感病毒的防御

• 批准号: 10537855
• 负责人: Jhonatan Henao Vasquez
• 金额: $ 4.68万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10537855
• 关键词: Address; Antiviral Agents; Area; Binding; Biology; Bone Marrow; Bone Marrow Transplantation; Cells; Cessation of life; Chimera organism; Critical Pathways; Data; Development; Dimensions; Disease; Effectiveness; Effector Cell; Epithelial Cells; Exhibits; Flow Cytometry; Foundations; Future; Goals; Growth; Harvest; Hematopoietic; Heterozygote; Homozygote; Host Defense; Immune; Immune response; Immunity; Infection; Inflammatory; Influenza; Influenza A Virus, H1N1 Subtype; Influenza A virus; Knock-in; Lung; Modeling; Mus; Nature; PTPRC gene; Pathway interactions; Patients; Play; Population; Process; Protein Family; Proteins; Public Health; Puerto Rico; RNA; Reporter; Research; Research Personnel; Respiratory Tract Infections; Retrotransposon; Role; Saline; Seasons; Sequence Alignment; Somatic Cell; Testis; Training; Tropism; Vaccines; Viral; Viral Pathogenesis; Virion; Virus Diseases; Wild Type Mouse; Work; airway epithelium; career; experimental study; flu; genome integrity; global health; high risk; immune function; immunoregulation; influenza infection; lung development; mouse model; novel; novel therapeutics; particle; response; single cell sequencing; single-cell RNA sequencing; transcriptomics; vaccine access; viral RNA

Abstract Influenza is one of the most common respiratory infections globally and has led to an alarming number of deaths annually. Seasonal vaccines and therapies vary in efficacy among higher risk patients although they currently serve as the only countermeasures. This presents a critical need to further understand the host response to influenza in order to uncover pathways that can be targeted using novel therapies. One of the target cells of Influenza A virus (IAV) are the multiciliated cells of the lung airway. Our lab had previously discovered a subset of multiciliated cells defined by the expression of MIWI2, an Argonaute family protein most commonly studied in mammalian testes. MIWI2 binds to PIWI-interacting RNA (piRNA) and suppresses retrotransposon activity in germline cells to maintain genome integrity. However, MIWI2 expression in somatic cells is not well understood. In order to understand lung MIWI2, we used a MIWI2-TdTomato heterozygote (haplo-sufficient) and a homozygote (deficient) knock-in reporter mouse model. Mice deficient in MIWI2 exhibited a markedly decrease burden of Influenza A virus suggesting that MIWI2 plays a critical role in regulating viral pathogenesis and the immune response. Incorporating an HA-mNeon reporter IAV, we ruled out the possibility that MIWI2 multiciliated cells were preferentially infected as we observed no differences in viral tropism between MIWI2 haplo-sufficient and deficient mice. Recently, we identified the presence of an immune cell population that also expresses MIWI2, raising the prospect that this heretofore unrecognized hematopoietic cell could be key to MIWI2 dependent immune function. Taken together, we hypothesize that a MIWI2 expressing cell in the lung critically modulates the host response to IAV infection. We will examine our central hypothesis by pursuing two aims. In the first Aim, we will use bone marrow transplantations and cell-specific deletion of MIWI2 mouse models to determine the identity of the MIWI2 expressing cell population that modulates the immune response during an IAV infection. In the second Aim, we will employ single cell sequencing to elucidate MIWI2-dependent pathways that critically regulate the lung’s antiviral immune response. Overall, the work proposed here will expand our understanding of the viral pathogenesis of IAV as well as uncover novel pathways that are involved in the lung’s host immune response. These studies will also provide functional information regarding the action of Argonaute proteins in somatic cells. Most importantly, this serves as a platform for an integrated and cogent training plan that will support my development as an independent researcher.

抽象的 流感是全球最常见的呼吸道感染之一，导致了令人震惊的死亡人数 每年。季节性疫苗和疗法在较高风险患者的效率上有所不同 作为唯一的对策​​。这提出了进一步理解主机回应的迫切需要 流感是为了发现可以使用新疗法来靶向的途径。一个目标细胞之一 流感病毒（IAV）是肺气道的多细胞细胞。我们的实验室以前发现了一个子集 由MIWI2表达定义的多细胞细胞，MIWI2的表达是一种最常见的argona家族蛋白蛋白 哺乳动物测试。 MIWI2与PIWI相互作用RNA（PIRNA）结合，并抑制反转跨活性 生殖细胞维持基因组完整性。但是，体细胞中的MIWI2表达尚不清楚。 为了了解肺Miwi2，我们使用了MIWI2-TDTOMATO杂合子（Haplo Wipplo）和A 纯合子（不足）敲入报告基因鼠标模型。缺乏MIWI2的小鼠显着下降了 流感的负担表明MIWI2在调节病毒发病机理和 免疫反应。结合了HA-MNEON的记者IAV，我们排除了MIWI2多智的可能性 细胞优选感染，因为我们观察到MIWI2单倍体充足的病毒性朝向主义没有差异 和缺乏小鼠。最近，我们确定了也表达的免疫菌株人群的存在 miwi2，提高了这种迄今未识别的造血细胞的前景可能是MIWI2的关键 依赖免疫功能。综上所述，我们假设一个MIWI2在肺中表达细胞 调节宿主对IAV感染的反应。我们将通过追求两个目标来研究我们的中心假设。在 第一个目的，我们将使用MIWI2小鼠模型的骨髓移植和细胞特异性缺失 确定表达细胞群的MIWI2的身份，该细胞群调节在 IAV感染。在第二个目标中，我们将采用单细胞测序来阐明MIWI2依赖性途径 这严重调节了肺部的抗病毒免疫反应。总体而言，这里提出的工作将扩大我们的 了解IAV的病毒发病机理以及发现与肺有关的新途径 宿主免疫反应。这些研究还将提供有关Argonaute行动的功能信息 体细胞中的蛋白质。最重要的是，这是整合培训计划的平台 这将支持我作为独立研究人员的发展。