Viral Hemorrhagic Fevers: Disease Modeling and Transmission

病毒性出血热：疾病建模和传播

基本信息

批准号：
10927843
负责人：
Heinrich Feldmann
金额：
$ 211.52万
依托单位：
NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10927843
关键词：
Acute Acute Respiratory Distress Syndrome Adrenal Glands African Alphavirus Animal Model Animals Antibody titer measurement Antigens Antiviral Agents Area Arenavirus Biological Assay Biology Brain Breeding Bunyavirales COVID-19 pandemic Case Fatality Rates Cells Cellular Immunity Characteristics Chiroptera Clinical Communicable Diseases Containment Control Groups Coronavirus Crimean-Congo Hemorrhagic Fever Virus DNA Data Development Diagnostic Diagnostic tests Disease Disease Outbreaks Disease Progression Disease model Dose Ebola Ebola Hemorrhagic Fever Ebola Vaccines Ebola virus Ecology Emergency Situation Emerging Communicable Diseases Euthanasia Eye Family Family suidae Ferrets Fever Filovirus Fostering Functional disorder Future Generations Genes Genetic Transcription Genome Geographic Distribution Geography Glycoproteins Goals Goat Hemorrhage Hour Human Humoral Immunities Immune response Immunity Immunization In Vitro Individual Infection International Intervention Intramuscular Intravenous Licensing Licensure Liver Livestock Lung Macaca Macaca fascicularis Macaca mulatta Marburg Virus Disease Marburgvirus Mastomys Membrane Glycoproteins MicroRNAs Microbe Modeling Molecular Mus Nanostructures National Institute of Allergy and Infectious Disease Neurologic Signs Nucleoproteins Organ Orthobunyavirus Outcome Pathogenesis Pathogenicity Pathology Plasmids Play Preventative vaccination Prophylactic treatment Public Health RNA RNA Editing RNA Interference RNA Viruses RNA amplification RNA vaccine Rattus Recombinants Recovery Regimen Replicon Reporting Research Research Activity Rest Rodent Role Sheep Spleen Structural Protein Sudan Ebola virus Syndrome System Technology Therapeutic Ticks Transcript Tropism Uganda United States National Institutes of Health Vaccinated Vaccination Vaccinee Vaccines Vesicular stomatitis Indiana virus Viral Viral Hemorrhagic Fevers Viral Pathogenesis Viral Proteins Viremia Virus Virus Diseases Virus Replication Western Blotting Work comparison control cross immunity cross reactivity disease transmission efficacy study epidemiology study experimental study exposure route immunogenicity improved in vivo in vivo evaluation innovation member mouse model neglect neglected tropical diseases nonhuman primate novel vaccines outbreak response pathogen pathogenic virus protective efficacy reproductive organ research facility response reverse genetics structured lipid success therapeutic development tick bite trait transmission process vaccine candidate vaccine development vaccine platform vector vector vaccine virus host interaction

项目摘要

During FY23 the focus of DMTs work was on filoviruses and orthonairoviruses. Filoviruses: Infections with ebolaviruses and marburgviruses, family Filoviridae, cause Ebola (EVD) and Marburg virus disease (MVD), respectively, with high case fatality rates. In addition, there are other filovirus genera, such as cuevaviruses, that have not been associated with human infections and disease. The natural reservoirs for filoviruses are likely different bat species. Filovirus are enveloped, non-segmented, negative-stranded RNA viruses expressing seven structural proteins. Some filoviruses, but not all, express nonstructural glycoproteins through RNA editing of the glycoprotein gene. The West African EVD outbreak has led to the licensure of vaccines treatments making EVD a success story in the field of neglected tropical diseases. (Marzi and Feldmann, J Infect Dis 2023) RNA editing has been discovered as an essential mechanism for the transcription of the glycoprotein (GP) gene of Ebola virus (EBOV). We developed a rapid transcript quantification assay (RTQA) to analyze RNA transcripts generated through RNA editing. RTQA successfully quantified the two major GP gene transcripts during infection with representative members of five ebolavirus species. Immunoblotting verified expression of the soluble (sGP) and the transmembrane GP. Our results defined RNA editing as a general trait of ebolaviruses. (Mehedi et al., J Infect Dis 2023) "Post-Ebola Syndrome" has first been described during and after the West African Ebola outbreak. In one of our studies a rhesus macaque developed neurological signs and acute respiratory distress following recovery of an Ebola virus (EBOV) infection requiring euthanasia. Interestingly, the organ tropism had changed with high virus titers in lungs, brain, eye, and reproductive organs but no virus in liver, spleen and adrenal glands, the typical target organs for acute EBOV infection. Thus, we have described an atypical EBOV disease similar to Post-Ebola Syndrome. It remains to be seen if the syndrome could be modeled and studied in future. (Marzi et al., J Infect Dis 2023) The focus of filovirus work during FY23 was on VSV-vectored filovirus vaccines. We generated and characterized VSV- SUDV and evaluated the protective efficacy following a single-dose vaccination against lethal SUDV infection in cynomolgus macaques. As we repurposed macaques from a successful previous VSV-Ebola virus (EBOV) vaccine efficacy study, we also investigated VSV-EBOV's cross-protective potential against SUDV challenge. Of the six NHPs given VSV-SUDV, none showed any signs of disease in response to the challenge. Four of the five NHPs in the control group developed characteristic clinical signs of SUDV diseases. Although the NHPs developed cross-reactive humoral responses to SUDV after VSV-EBOV vaccination and EBOV challenge, there was little cross-protection. These data emphasize the need for species-specific vaccines for ebolaviruses. Although previous VSV-EBOV immunity is boosted through VSV-SUDV vaccination, it has only limited effect on the immunogenicity and protective efficacy of VSV-SUDV. This study presented a milestone study informing public health during the SUDV outbreak in Uganda. Marzi et al, Lancet Microbe 2023 Ebola virus (EBOV) and Marburg virus (MARV) have overlapping endemicity areas. We used macaques previously vaccinated with VSV-MARV and protected against MARV challenge. After a resting period of 9 months, macaques were re-vaccinated with VSV-EBOV and challenged with EBOV resulting in 75% survival. Surviving NHPs developed EBOV GP-specific antibody titers and no viremia or clinical signs. This study again demonstrates that VSVG-based filovirus vaccine can be successfully used in individuals with pre-existing VSV vector immunity highlighting the platform's applicability for consecutive outbreak response. (Marzi et al., J Infect Dis 2023) VSV-EBOV has been successfully used in ring vaccination approaches during Ebola disease outbreaks demonstrating its general benefit in short-term prophylactic vaccination, but actual proof of its benefit in true post-exposure prophylaxis (PEP) for humans is missing. Animal studies have indicated PEP efficacy when VSV-EBOV was used within hours of lethal EBOV challenge. Here, we used a lower EBOV challenge dose and a combined intravenous and intramuscular VSV-EBOV administration to improve PEP efficacy in the rhesus macaque model. VSV-EBOV treatment 1 hour post EBOV challenge resulted in delayed disease progression but little benefit in outcome. Thus, we could not confirm previous results indicating questionable benefit of VSV-EBOV for EBOV PEP even in a nonhuman primate model. (Bushmaker et al., J Infect Dis 2023) Although significant progress has been made in the development of therapeutics against EBOV, we sought to expand upon existing strategies and combine an RNAi-based intervention with the approved VSV-EBOV vaccine to conjointly treat and vaccinate patients during an outbreak. We constructed VSV-EBOV vectors expressing artificial miRNAs (amiRNAs) targeting sequences of EBOV proteins. In vitro experiments demonstrated a robust decrease in EBOV replication using a minigenome system and infectious virus. For in vivo evaluation, MA-EBOV-infected CD-1 mice were treated 24 hours after infection with a single dose of the VSV-EBOV-amiRNA constructs. We observed no difference in disease progression or survival compared to the control-treated mice. In summary, while amiRNAs decrease viral replication in vitro, the effect is not sufficient to protect mice from lethal disease, and this therapeutic approach requires further optimization. (ODonnell et al., J Infect Dis 2023) Orthonairoviruses Introduction: Crimean-Congo hemorrhagic fever virus (CCHFV) is a tri-segmented, negative sense virus in the Bunyavirales order. The principal vector and reservoir are ticks of the Hyalomma genus and the wide geographic distribution of CCHFV follows the geographic range of these ticks. The primary routes of exposure are tick-bites and handling of infected livestock, although human-to-human spread is reported. Clinically, CCHF presents initially as a non-specific febrile illness that can rapidly progress to a severe, sometimes fatal hemorrhagic disease. The host and viral determinants of CCHFV pathogenesis are poorly understood and there are no approved vaccines or antivirals. (Hawman and Feldmann, Nat Rev Microbiol 2023) We have previously reported significant efficacy of a three-dose DNA-based vaccination regimen for CCHFV in cynomolgus macaques (Hawman et al., Nat Microbiol 2021). Here, we show that in macaques, plasmid expressed CCHFV nucleoprotein (NP) and glycoprotein precursor (GPC) antigens elicit primarily humoral and cellular immunity, respectively. We found that a two-dose vaccination regimen with plasmids expressing the NP and GPC provides significant protection against CCHFV infection. Studies investigating vaccinations with either antigen alone showed that plasmid-expressed NP could also confer protection. Our data show that this vaccine confers robust protection against CCHFV. (Hawman et al., Mol Ther 2023) We evaluated an alphavirus-based replicon RNA vaccine expressing either the CCHFV NP or GPC in a heterologous lethal mouse model. Vaccination with the RNA expressing NP alone could confer complete protection against clinical disease, but vaccination with a combination of both the NP and GPC afforded robust protection against disease and viral replication. This vaccine conferred robust protection against CCHFV and supports continued development of this vaccine. (Leventhal et al., EBioMedicine 2022)

2023 财年，DMT 的工作重点是丝状病毒和正呼吸道病毒。丝状病毒：埃博拉病毒和马尔堡病毒（丝状病毒科）的感染分别导致埃博拉病毒（EVD）和马尔堡病毒病（MVD），病死率很高。此外，还有其他丝状病毒属，例如提示病毒属，与人类感染和疾病无关。丝状病毒的天然宿主可能是不同的蝙蝠物种。丝状病毒是有包膜、不分段、负链 RNA 病毒，表达七种结构蛋白。一些丝状病毒（但不是全部）通过糖蛋白基因的 RNA 编辑表达非结构糖蛋白。西非埃博拉病毒病疫情导致疫苗治疗获得许可，使埃博拉病毒病成为被忽视的热带疾病领域的成功案例。（Marzi 和 Feldmann，J Infect Dis 2023） RNA 编辑已被发现是埃博拉病毒 (EBOV) 糖蛋白 (GP) 基因转录的重要机制。我们开发了一种快速转录本定量分析 (RTQA) 来分析通过 RNA 编辑生成的 RNA 转录本。 RTQA 成功量化了五种埃博拉病毒代表性成员感染期间两个主要 GP 基因转录本。免疫印迹验证了可溶性 (sGP) 和跨膜 GP 的表达。我们的结果将 RNA 编辑定义为埃博拉病毒的一般特征。（Mehedi 等人，J Infect Dis 2023） “后埃博拉综合症”是在西非埃博拉疫情期间和之后首次被描述的。在我们的一项研究中，一只恒河猴在埃博拉病毒 (EBOV) 感染恢复后出现了神经系统症状和急性呼吸窘迫，需要安乐死。有趣的是，器官向性发生了变化，肺、脑、眼和生殖器官中的病毒滴度很高，但肝、脾和肾上腺（急性埃博拉病毒感染的典型靶器官）中没有病毒。因此，我们描述了一种类似于埃博拉后综合症的非典型埃博拉病毒疾病。未来是否可以对这种综合症进行建模和研究还有待观察。（Marzi 等人，J Infect Dis 2023） 2023 财年丝状病毒工作的重点是 VSV 载体丝状病毒疫苗。我们生成并表征了 VSV-SUDV，并评估了食蟹猴单剂疫苗接种后针对致命性 SUDV 感染的保护功效。当我们重新利用先前成功的 VSV-埃博拉病毒 (EBOV) 疫苗功效研究中的猕猴时，我们还研究了 VSV-EBOV 针对 SUDV 攻击的交叉保护潜力。在接受 VSV-SUDV 治疗的 6 名 NHP 中，没有人因挑战而表现出任何疾病迹象。对照组的 5 名 NHP 中有 4 名出现了 SUDV 疾病的特征性临床症状。尽管 NHP 在 VSV-EBOV 疫苗接种和 EBOV 攻击后对 SUDV 产生了交叉反应性体液反应，但几乎没有交叉保护。这些数据强调了对埃博拉病毒物种特异性疫苗的需求。虽然之前通过VSV-SUDV疫苗接种增强了VSV-EBOV免疫力，但对VSV-SUDV的免疫原性和保护功效作用有限。这项研究提出了一项里程碑式的研究，为乌干达 SUDV 爆发期间的公共卫生提供信息。 Marzi 等人，柳叶刀微生物 2023 埃博拉病毒（EBOV）和马尔堡病毒（MARV）有重叠的流行区域。我们使用之前接种过 VSV-MARV 疫苗并免受 MARV 攻击的猕猴。经过 9 个月的休息期后，猕猴重新接种 VSV-EBOV 疫苗并接受 EBOV 攻击，存活率达到 75%。幸存的 NHP 出现了 EBOV GP 特异性抗体滴度，并且没有病毒血症或临床症状。这项研究再次表明，基于 VSVG 的丝状病毒疫苗可以成功用于已有 VSV 载体免疫力的个体，凸显了该平台对连续疫情应对的适用性。（Marzi 等人，J Infect Dis 2023） VSV-EBOV 已在埃博拉疾病爆发期间成功用于环形疫苗接种方法，证明了其在短期预防性疫苗接种中的普遍益处，但缺乏其对人类真正的暴露后预防 (PEP) 益处的实际证据。动物研究表明，在致死性 EBOV 攻击后数小时内使用 VSV-EBOV 时，PEP 具有功效。在这里，我们使用较低的 EBOV 攻击剂量以及静脉内和肌肉内联合注射 VSV-EBOV 来提高恒河猴模型中的 PEP 疗效。 EBOV 攻击后 1 小时进行 VSV-EBOV 治疗可延迟疾病进展，但对结果几乎没有好处。因此，我们无法证实之前的结果表明，即使在非人类灵长类动物模型中，VSV-EBOV 对 EBOV PEP 的益处也存在疑问。（Bushmaker 等人，J Infect Dis 2023）尽管在开发针对 EBOV 的疗法方面取得了重大进展，但我们试图扩展现有策略，并将基于 RNAi 的干预措施与已批准的 VSV-EBOV 疫苗相结合，以便在疫情爆发期间联合治疗和为患者接种疫苗。我们构建了表达针对 EBOV 蛋白序列的人工 miRNA (amiRNA) 的 VSV-EBOV 载体。体外实验表明，使用微型基因组系统和传染性病毒可显着减少埃博拉病毒复制。为了进行体内评估，感染 MA-EBOV 的 CD-1 小鼠在感染后 24 小时用单剂量的 VSV-EBOV-amiRNA 构建体进行治疗。与对照组治疗的小鼠相比，我们观察到疾病进展或存活率没有差异。综上所述，虽然amiRNA在体外减少病毒复制，但其效果不足以保护小鼠免受致命性疾病，这种治疗方法需要进一步优化。（ODonnell 等人，J Infect Dis 2023）正呼吸道病毒属简介：克里米亚-刚果出血热病毒（CCHFV）是布尼亚病毒目中的一种三节段负义病毒。主要传播媒介和宿主是玻璃蜱属蜱，CCHFV 的广泛地理分布遵循这些蜱的地理范围。尽管有报道称存在人际传播，但主要的接触途径是蜱虫叮咬和处理受感染的牲畜。临床上，CCHF 最初表现为一种非特异性发热性疾病，可迅速进展为严重的、有时致命的出血性疾病。人们对 CCHFV 发病机制的宿主和病毒决定因素知之甚少，并且没有批准的疫苗或抗病毒药物。（Hawman 和 Feldmann，Nat Rev Microbiol 2023）我们之前曾报道过基于 DNA 的三剂疫苗接种方案对食蟹猴 CCHFV 具有显着疗效（Hawman 等人，Nat Microbiol 2021）。在这里，我们表明，在猕猴中，质粒表达的 CCHFV 核蛋白（NP）和糖蛋白前体（GPC）抗原分别主要引发体液免疫和细胞免疫。我们发现，使用表达 NP 和 GPC 的质粒进行两剂疫苗接种方案可提供针对 CCHFV 感染的显着保护。单独使用任一抗原进行疫苗接种的研究表明，质粒表达的 NP 也可以提供保护。我们的数据表明，这种疫苗可以针对 CCHFV 提供强有力的保护。（Hawman 等人，Mol Ther 2023）我们在异源致死小鼠模型中评估了表达 CCHFV NP 或 GPC 的基于甲病毒的复制子 RNA 疫苗。单独使用表达 NP 的 RNA 进行疫苗接种可以提供针对临床疾病的完全保护，但同时使用 NP 和 GPC 进行疫苗接种可以提供针对疾病和病毒复制的强大保护。该疫苗对 CCHFV 具有强大的保护作用，并支持该疫苗的持续开发。（Leventhal 等人，EBioMedicine 2022）

项目成果

期刊论文数量（186）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

The Ebola virus glycoprotein and HIV-1 Vpu employ different strategies to counteract the antiviral factor tetherin.

DOI：
10.1093/infdis/jir378
发表时间：
2011-11
期刊：
The Journal of infectious diseases
影响因子：
0
作者：
Kühl A;Banning C;Marzi A;Votteler J;Steffen I;Bertram S;Glowacka I;Konrad A;Stürzl M;Guo JT;Schubert U;Feldmann H;Behrens G;Schindler M;Pöhlmann S
通讯作者：
Pöhlmann S

Single-Nucleotide Polymorphisms in Human NPC1 Influence Filovirus Entry Into Cells