Host and Viral Determinants of Orthobunyavirus Vertical Transmission: Novel Model Systems to Understand the Mechanisms of Congenital Disease in Humans and Ruminants

正布尼亚病毒垂直传播的宿主和病毒决定因素：了解人类和反刍动物先天性疾病机制的新模型系统

• 批准号: 10589470
• 负责人: Cynthia McMillen
• 金额: $ 12.48万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-01 至 2028-07-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10589470
• 关键词: Affect; Agriculture; Antiviral Response; Apoptosis; Apoptotic; Arboviruses; Autophagocytosis; Biological Models; Biological Response Modifiers; Birth; Bunyavirus Infections; COVID-19 pandemic; Cache Valley virus; Cell Death; Cells; Cellular Tropism; Communities; Containment; Culicidae; Data; Development; Development Plans; Disease; Economics; Emerging Communicable Diseases; Environment; Event; Fetal Death; Gene Expression; Generations; Genetic; Goals; Health; Human; Immune response; Immunology; In Vitro; Infection; Inhibition of Apoptosis; Innate Immune Response; Integration Host Factors; Interferon Type I; Interferons; Investigation; Knock-out; La Crosse virus; Laboratories; Life; Livestock; Maternal-Fetal Exchange; Mediating; Mediator; Medical; Mentors; Mentorship; Modeling; Natural Immunity; Nature; Nonstructural Protein; Organoids; Orthobunyavirus; Pathogenesis; Pathology; Pathway interactions; Placenta; Placental Biology; Plants; Predisposition; Pregnant Women; Process; Productivity; Proteins; Rattus; Research; Rift Valley Fever; Rift Valley fever virus; Ruminants; Scientist; Severities; Sheep; Spontaneous abortion; Syncytiotrophoblast; System; Teratogens; Tissues; Training; Tropism; Universities; Vertical Transmission; Viral; Viral Pathogenesis; Virulence Factors; Virus; Virus Diseases; Virus Replication; animal facility; career; career development; congenital infection; experience; fetal; high risk; laboratory facility; multiple myeloma M Protein; novel; permissiveness; placental infection; pregnant; prevent; programs; prototype; responsible research conduct; tissue tropism; training opportunity; transmission process; trophoblast; zoonotic spillover

PROJECT SUMMARY/ABSTRACT Emerging infectious diseases pose a significant threat to human and agricultural health; therefore, it is imperative that we take a proactive approach toward understanding virus and host factors that are associated with infection and pathogenesis. Many bunyavirus infections, including Rift Valley fever (RVFV), Cache Valley (CVV), and Schmallenberg (SBV) viruses, cause massive abortogenic events in livestock that lead to significant economic strain and increased susceptibility of human infection. La Crosse virus (LCV), a related bunyavirus, is not known to cause vertical transmission in livestock, however vertical transmission has been implicated upon experimental infection. Two cases of vertical transmission of RVFV have occurred in pregnant women and those infected with RVFV have a higher risk for late-term miscarriages. Whether vertical transmission occurs in ruminants and humans due to LACV infection or in humans from SBV and CVV infection is unknown. Given LACV and CVV infections can cause life-threatening diseases in humans, it is plausible that congenital infection may simply be overlooked due to the mild nature of most bunyavirus infections. We hypothesize that SBV, CVV, and LACV can infect human and ruminant placentas and that virulence factors, such as NSm, and variable induction of antiviral responses across viruses dictate pathogenesis severity, and thus teratogenicity, across host species. This proposal will utilize two model systems to study bunyavirus infection of the placenta. First, we will examine whether LACV, CVV, SBV, and RVFV infect placenta explants from humans, sheep, and rats in vitro. Using wild type and NSm knockout viruses, we will identify whether NSm contributes to host or cellular tropism, immune responses to infection, programmed cell death pathways, and congenital pathogenesis. Second, we will utilize genetically tractable 2D human trophoblast and trophoblast organoid (TO) systems to compare cell- specific differences in immune responses and cell death pathways upon bunyavirus infection. This will be the first study to utilize human placenta organoids to study congenital bunyavirus infection. To successfully complete the proposed project, I have developed an exceptional career development plan under the primary mentorship of Dr. Amy Hartman (University of Pittsburgh (U Pitt)) and co-mentors, Drs. Carolyn Coyne (Duke University) and Leonard D’Aiuto (U Pitt). My training will consist of hands-on and didactic training in human organoid development in addition to didactic training in pathology, cross-species placenta biology, and immunology. U Pitt provides the necessary environment to support the proposed research and training through accessibility to outstanding scientists, high containment laboratory and animal facilities, training opportunities in laboratory management, responsible conduct of research and grantsmanship, and opportunities to present research. Successful training and completion of the proposed research will support my ultimate career goal to establish an independent research program studying the cross virus- and host-species mechanisms of bunyavirus congenital infections with a special niche in human and ruminant placenta organoid research and congenital pathology.

项目摘要/摘要 新出现的传染病对人类和农业健康构成重大威胁;因此， 我们应该采取积极主动的方法来了解病毒和与感染相关的宿主因素 和发病机制。许多布尼亚病毒感染，包括裂谷热（RVFV），Cache Valley（CVV）， Schmallenberg（SBV）病毒在家畜中引起大规模的流产事件，导致重大的经济损失。 菌株和增加人类感染易感性。拉克罗斯病毒（LCV），一种相关的布尼亚病毒，尚不清楚 在家畜中引起垂直传播，然而，在实验中发现了垂直传播， 感染两例RVFV垂直传播发生在孕妇和感染RVFV的人中。 RVFV晚期流产的风险更高。反刍动物是否发生垂直传播， 在人类中由于LACV感染或在人类中由于SBV和CVV感染是未知的。给予LACV， CVV感染可导致危及人类生命的疾病，先天性感染可能只是 由于大多数布尼亚病毒感染的性质温和，因此被忽视。我们假设SBV、CVV和LACV 可以感染人和反刍动物胎盘，毒力因子，如NSm，和可变的诱导 跨病毒的抗病毒应答决定了跨宿主物种的发病机制严重性，从而决定了致畸性。 本提案将利用两个模型系统来研究布尼亚病毒感染的胎盘。一是 在体外检测LACV、CVV、SBV和RVFV是否感染来自人、绵羊和大鼠的胎盘外植体。 使用野生型和NSm敲除病毒，我们将鉴定NSm是否有助于宿主或细胞向性， 对感染的免疫反应、程序性细胞死亡途径和先天性发病机制。二是 将利用遗传学上易于处理的2D人类滋养层和滋养层类器官（TO）系统来比较细胞- 布尼亚病毒感染后免疫应答和细胞死亡途径的特异性差异。这将是 第一项利用人类胎盘类器官研究先天性布尼亚病毒感染的研究。顺利完成 在建议的项目中，我已经制定了一个特殊的职业发展计划， Amy Hartman博士（匹兹堡大学（U Pitt））和共同导师Carolyn Coyne博士（杜克大学） 还有伦纳德·达伊托（U·皮特饰）。我的培训将包括人类类器官的实践和教学培训 除了病理学、跨物种胎盘生物学和免疫学方面的教学培训外，还提供发展方面的培训。U皮特 提供必要的环境，以支持拟议的研究和培训， 杰出的科学家，高封闭实验室和动物设施，实验室培训机会 管理，负责任地进行研究和资助，并有机会提出研究。 成功的培训和完成拟议的研究将支持我的最终职业目标，建立一个 研究先天性本扬病毒跨病毒和宿主种机制的独立研究计划 在人类和反刍动物胎盘类器官研究和先天性病理学中具有特殊生态位的感染。