Characterizing the role of LDL related receptor 1 (Lrp1) as host entry factor for multiple bunyaviruses

项目摘要/摘要 布尼亚病毒(布尼亚病毒目：布尼亚病毒目)是一种日益增长和多样化的动物和人类病原体家族，具有 大流行的可能性。拥有300多名成员，蚊子和扁虱媒介的分布不断扩大，原因是 气候变化，这些病毒是导致人类疾病暴发增加的原因，并呈现出 对人类健康构成重大威胁。裂谷热病毒(RVFV)是目前研究较多的本雅热病毒之一。 被指定为NIAID A类病原体，并包括在世卫组织的优先疾病蓝图中。这个 防疫创新联盟(CEPI)将RVFV作为其新出现的传染病的一部分 疾病疫苗计划，进一步强调了对全球健康和经济的潜在影响。 奥罗波什病毒(Oropouche Virus，OROV)发现于南美洲，曾引发30多起大规模疫情，导致 超过50万人感染，使其成为南美洲仅次于 登革热。然而，由于奥罗波什热被误诊，真实病例数量可能要高得多。 基孔肯雅或登革热。第三个成员，La Crosse病毒(LACV)主要在北美发现， 在美国，儿童病毒性脑炎的主要原因。无论是奥罗夫还是拉科夫都不是 被研究为RVFV，因此在我们对本雅氏病毒致病机制的广泛理解中仍然存在着显著的差距。 目前还没有批准的治疗药物来治疗RVFV、OROV或LACV病，进一步 突出了我们建议进行研究的必要性。为了满足这一需求，我们进行了基因组筛查 确定了几个关键因素，包括低密度脂蛋白家族成员LRP1。在支持方面，我们提供令人信服的 初步数据包括LRP1充足和缺陷细胞的体外验证、反互补研究、 与RVFV糖蛋白Gn在体外直接相互作用。我们还发现内源对LRP1的抑制 来自进化不同宿主的多个细胞系的体外配体，以及体内数据显示 LRP1在小鼠的病毒嗜性和疾病中的重要性。在这里，我们将描述LRP1对以下方面的重要性 多种布尼亚病毒的进入，确定分子机制，并在体外和体内验证其意义。 这项工作将由具有各方面专业知识的高生产率和协作性的调查人员进行 包括生物化学、病毒发病机制、免疫学、蛋白质组学、结构生物学、 和病毒学。完成后，我们预计将验证LRP1作为泛布尼亚病毒的进入因素，填补 并为治疗开发提供新的靶点。

Project Summary/Abstract Bunyaviruses (Order: Bunyavirales) are a growing and diverse family of animal and human pathogens with pandemic potential. With over 300 members and an expanding distribution of mosquito and tick vectors due to climate change, these viruses are responsible for increasing outbreaks of human disease and present a significant threat to human health. Rift Valley Fever virus (RVFV) is one of the well-studied bunyaviruses and is designated as an NIAID Category A pathogen and included in the WHO’s Blueprint of Priority Diseases. The Coalition for Epidemic Preparedness Innovations (CEPI) included RVFV as part of their emerging infectious diseases vaccine program, further emphasizing the potential impact on the global health and economy. Oropouche virus (OROV) is found in South America and has caused more than 30 large epidemics resulting in over 500,000 human cases, making it the second most common arboviral disease in South America behind Dengue fever. However, the true case number is likely much higher due to Oropouche fever being misdiagnosed as Chikungunya or Dengue. A third member, La Crosse virus (LACV) is found primarily in North America and is the primary cause of pediatric viral encephalitis in the United States. Neither OROV nor LACV have been as well studied as RVFV, and thus a significant gap remains in our broad understanding of bunyavirus pathogenesis. Currently there are no approved therapeutic drugs for treatment of RVFV, OROV, or LACV disease, further highlighting the need for our proposed studies. To address this need, we conducted a genomic screen that defined several critical factors, including Lrp1, an LDL family member. In support we provide compelling preliminary data including in vitro validation in Lrp1 sufficient and deficient cells, transcomplementation studies, and direct interaction between RVFV glycoprotein Gn in vitro. We also show that inhibition of Lrp1 by endogenous ligands in vitro in multiple cell lines from evolutionarily distinct hosts, and in vivo data demonstrating the importance of Lrp1 for viral tropism and disease in mice. Here we will characterize the importance of Lrp1 for entry of multiple bunyaviruses, define molecular mechanisms, and validate the significance in vitro and in vivo. This work will be performed by highly productive and collaborative investigators with expertise in every aspect of the proposed studies, including biochemistry, viral pathogenesis, immunology, proteomics, structural biology, and virology. At completion, we expect to validate Lrp1 as pan-bunyavirus entry factor, filling a key gap in the field and to provide novel targets for therapeutic development.