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

基本信息

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
项目类别：
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

项目摘要

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.

抽象的流感是最常见的呼吸道感染之一，已导致全球死亡人数惊人每年，季节性疫苗和疗法在高风险患者中的功效各不相同，尽管目前它们是这样的。作为唯一的对策，这迫切需要进一步了解主机的反应。流感，以便发现可以使用新疗法靶向的途径。甲型流感病毒 (IAV) 是肺气道的多纤毛细胞，我们的实验室之前发现了一个子集。由 MIWI2 的表达定义的多纤毛细胞，MIWI2 是一种最常研究的 Argonaute 家族蛋白 MIWI2 与 PIWI 相互作用 RNA (piRNA) 结合并抑制逆转录转座子活性。然而，MIWI2 在体细胞中的表达尚不清楚。为了了解肺 MIWI2，我们使用了 MIWI2-TdTomato 杂合子（单倍体）和 MIWI2 缺陷的纯合子（缺陷）敲入报告小鼠模型显示出显着下降。甲型流感病毒的负担表明 MIWI2 在调节病毒发病机制和结合 HA-mNeon 报告基因 IAV，我们排除了 MIWI2 多纤毛的可能性。细胞优先被感染，因为我们观察到 MIWI2 单倍体之间的病毒向性没有差异最近，我们发现了也表达该基因的免疫细胞群的存在。 MIWI2，提出了这种迄今为止未被识别的造血细胞可能是 MIWI2 关键的前景综上所述，我们与肺部的 MIWI2 表达细胞进行了激烈的斗争。调节宿主对 IAV 感染的反应我们将通过追求两个目标来检验我们的中心假设。第一个目标，我们将利用骨髓移植和细胞特异性删除MIWI2小鼠模型来确定在免疫过程中调节免疫反应的 MIWI2 表达细胞群的身份在第二个目标中，我们将采用单细胞测序来阐明 MIWI2 依赖性途径。总的来说，这里提出的工作将扩大我们的研究范围。了解 IAV 的病毒发病机制以及涉及肺部疾病的新发现途径这些研究还将提供有关 Argonaute 作用的功能信息。最重要的是，这可以作为综合且令人信服的训练计划的平台。这将支持我作为一名独立研究员的发展。