Disease Modeling of Influenza and Other Emerging Respiratory Viral Pathogens

流感和其他新出现的呼吸道病毒病原体的疾病模型

• 批准号: 10272159
• 负责人: Heinrich Feldmann
• 金额: $ 85.65万
• 依托单位: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
• 依托单位国家: 美国
• 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/10272159
• 关键词: 2019-nCoV; Acute; African Green Monkey; Animal Disease Models; Animal Diseases; Animal Model; Animals; Antibodies; Antibody Response; Antiviral Agents; Avian Influenza; Avian Influenza A Virus; Bangladesh; COVID-19; Callithrix jacchus jacchus; Cessation of life; Clinical; Clinical Trials; Collaborations; Combined Modality Therapy; Containment; Control Animal; Coronavirus Infections; Data; Development; Disease; Disease Outbreaks; Disease model; Dose; Drug Screening; Emergency Situation; Emerging Communicable Diseases; Epidemic; Fostering; Functional disorder; Funding; GTP-Binding Proteins; Genotype; Goals; H5 influenza virus; Hantavirus; Hantavirus Pulmonary Syndrome; Hemagglutinin; Hendra Virus; Henipavirus; Human; Hydroxychloroquine; Immune response; Immunoglobulin G; In Vitro; Infection; Influenza; Influenza A Virus, H5N1 Subtype; Influenza A virus; International; Intramuscular Injections; Kinetics; Lead; Length; Lesion; Lipids; Lower respiratory tract structure; Lung; Lung diseases; Macaca fascicularis; Macaca mulatta; Malaria; Malaysia; Manuscripts; Mesocricetus auratus; Middle East Respiratory Syndrome; Middle East Respiratory Syndrome Coronavirus; Modeling; Monoclonal Antibodies; Mus; National Institute of Allergy and Infectious Disease; Nipah Virus; Oseltamivir; Outcome; Pathogenesis; Pathogenicity; Pathologic; Patients; Preparation; Prophylactic treatment; Proteins; Public Health; RNA; Recombinants; Replicon; Research; Resistance; SARS coronavirus; Severities; T cell response; Testing; Therapeutic; Tissues; Treatment Efficacy; Treatment Protocols; United States National Institutes of Health; Vaccinated; Vaccination; Vaccines; Vesicular stomatitis Indiana virus; Viral; Virulence; Virulent; Virus; Virus Diseases; Virus Replication; Wit; Work; anti-viral efficacy; antimicrobial; base; burden of illness; coronavirus disease; design; disease transmission; drug efficacy; efficacy testing; human pathogen; immunogenic; immunogenicity; in vivo evaluation; influenza virus vaccine; influenzavirus; innovation; inorganic phosphate; intravenous administration; mutant; nanoparticle; nonhuman primate; novel; pandemic disease; pathogenic virus; pre-clinical; prevent; prophylactic; remdesivir; respiratory; response; vaccination strategy; vaccine candidate; vaccine response; vector; vector vaccine

Over the past fiscal year, we have performed studies on four viral pathogens causing respiratory disease in humans, Nipah virus, influenza virus, Middle East Respiratory Syndrome coronavirus (MERS-CoV) and Severe Acute Respiratory Syndrome coronavirus 2 (SARS-CoV-2). From February to July 2020, only work on SARS-CoV-2 was permitted. (1)Nipah virus: We have continued work on two main projects, efficacy of the antiviral drug remdesivir and prophylactic efficacy of the VSV-based vaccine (VSV-Nipah) against Nipah virus infection. We tested the efficacy of remdesivir against Nipah virus (Bangladesh genotype) in African green monkeys. In contrast to control animals, which all succumbed to the infection, all GS-5734-treated animals survived the lethal challenge, indicating that GS-5734 is a promising antiviral treatment for Nipah virus infection (Lo et al. Sci Transl Med 2019). We also tested the efficacy of VSV-Nipah in African green monkeys against Nipah virus (Malaysia and Bangladesh genotypes) challenge. In contrast to control animals, which all succumbed, all vaccinated animals were completely protected against both challenges. The vaccine expressing the Nipah virus G protein performed slightly better in regard to the immunological responses than that vaccine expressing the Nipah virus F protein (de Wit et al., manuscript in preparation). VSV-Nipah (expressing G protein) will now be evaluated under CEPI funding in collaboration with Public Health Vaccines and moved into clinical trials. (2)Influenza virus: The observed virulence of the 1918 influenza virus substantiates the need for evaluating antiviral options against infections with this virulent human pathogen. Here we demonstrate that oseltamivir phosphate is effective in preventing severe disease in cynomolgus macaques if given prophylactically. Efficacy was reduced in a treatment regimen through emergence of oseltamivir-resistant mutants that lead to death of one of four animals. This emphasizes the importance of implementing combination therapy and vaccination strategies early in an epidemic or pandemic. (Feldmann et al., mBio 2019). The avian influenza virus outbreak in 1997 highlighted the potential of the highly pathogenic H5N1 virus to cause severe disease in humans. Therefore, effective vaccines against H5N1 viruses are needed to counter the potential threat of a global pandemic. We generated recombinant VSV-based H5N1 influenza virus vectors to demonstrate the feasibility of this platform for a fast-acting pan-H5 influenza virus vaccine. We found that a single dose of VSV vectors expressing full-length hemagglutinin (HAfl) were sufficient to provide 100% protection. The vaccine vectors were fast-acting as demonstrated by uniform protection when administered 3 days prior to lethal challenge. Moreover, single vaccination induced cross-protective H5-specific antibodies and protected mice against lethal challenge with various H5 clade 2 viruses, highlighting the potential of the VSV-based HAfl as a pan-H5 influenza virus emergency vaccine. (3)Middle East Respiratory Syndrome coronavirus (MERS-CoV): We performed two major therapeutic projects, efficacy of antibody and remdesivir treatment. We tested the prophylactic efficacy of the monoclonal antibody LCA60 in the common marmoset model of MERS-CoV infection. Intravenous administration of LCA60 one day before virus challenge resulted in a moderate clinical benefit of treatment including reduced respiratory involvement. Although viral lung loads were not reduced in LCA60-treated animals as compared to controls, there were fewer pathological changes in the lungs. Thus, prophylactic LCA60 treatment could be implemented to reduce disease burden in contacts of confirmed MERS-CoV patients. Furthermore, we tested the efficacy of prophylactic and therapeutic remdesivir treatment in the rhesus macaque model of MERS-CoV infection. Prophylactic remdesivir treatment initiated 24 h prior to inoculation completely prevented MERS-CoV-induced clinical disease, strongly inhibited MERS-CoV replication in respiratory tissues, and prevented the formation of lung lesions. Therapeutic remdesivir treatment initiated 12 h post inoculation also provided a clear clinical benefit, with a reduction in clinical signs, reduced virus replication in the lungs, and decreased presence and severity of lung lesions. The data presented here support testing of the efficacy of remdesivir treatment in the context of a MERS clinical trial. (4)Severe Acute Respiratory Syndrome coronavirus (SARS-CoV-2): Since February 2020 we have been mainly working on COVID projects. We have established the Syrian hamster disease model for pathogenesis studies and countermeasure development (Rosenke et al., manuscript in preparation). We have established an in vitro drug screening pipeline that will feed promising candidates into in vivo testing using the Syrian hamster model (Jarvis et al., Antivir Ther 2020; Rosenke et al., J Antimicrobial Chemotherap, submitted). These studies are ongoing. We further assessed the prophylactic/therapeutic efficacy of hydroxychloroquine (HCQ) in two animal disease models. The standard human malaria HCQ prophylaxis and treatment did not significantly benefit clinical outcome nor reduce SARS-CoV-2 replication/shedding in the upper and lower respiratory tract in the rhesus macaque disease model. Similarly, when used for prophylaxis or treatment neither the standard human malaria dose nor a high dose of HCQ had any beneficial effect on clinical disease or SARS-CoV-2 kinetics (replication/shedding) in the Syrian hamster disease model. Results from these two preclinical animal models may prove helpful in guiding clinical use of HCQ for prophylaxis/treatment of COVID-19. Finally, we have rapidly developed repRNA-CoV2S, a stable and highly immunogenic vaccine candidate comprised of an RNA replicon formulated with a novel Lipid InOrganic Nanoparticle (LION) designed to enhance vaccine stability, delivery, and immunogenicity. We have shown that intramuscular injection of LION/repRNA-CoV2S elicited robust anti-SARS-CoV-2 spike protein IgG antibody isotypes indicative of a Type 1 T helper response as well as potent T cell responses in mice. Importantly, a prime-only administration in nonhuman primates elicited antibody responses that potently neutralized SARS-CoV-2 as well as T cell responses indicative of a Type 1 T helper response. These data support further development of LION/repRNA-CoV2S as a vaccine candidate for prophylactic protection from SARS-CoV-2 infection (Erasmus et al., Sci Transl Med 2020).

在过去的财年中，我们对四种引起人类呼吸道疾病的病毒病原体进行了研究：尼帕病毒、流感病毒、中东呼吸综合征冠状病毒（MERS-CoV）和严重急性呼吸综合征冠状病毒2（SARS-CoV-2）。从 2020 年 2 月到 7 月，只允许进行 SARS-CoV-2 的研究。 （1）尼帕病毒：我们继续开展两个主要项目的研究，抗病毒药物瑞德西韦的功效和基于VSV的疫苗（VSV-Nipah）对尼帕病毒感染的预防功效。 我们在非洲绿猴身上测试了瑞德西韦对抗尼帕病毒（孟加拉国基因型）的功效。与全部死于感染的对照动物相比，所有 GS-5734 治疗的动物都在致命挑战中幸存下来，这表明 GS-5734 是一种有前途的尼帕病毒感染抗病毒治疗方法 (Lo et al. Sci Transl Med 2019)。我们还在非洲绿猴中测试了 VSV-Nipah 对抗尼帕病毒（马来西亚和孟加拉国基因型）挑战的功效。与全部死亡的对照动物相比，所有接种疫苗的动物都完全免受这两种挑战。表达尼帕病毒 G 蛋白的疫苗在免疫反应方面比表达尼帕病毒 F 蛋白的疫苗表现稍好（de Wit 等人，手稿正在准备中）。 VSV-Nipah（表达 G 蛋白）现将在 CEPI 资助下与公共卫生疫苗公司合作进行评估，并进入临床试验。 (2) 流感病毒：观察到的 1918 年流感病毒的毒力证实有必要评估针对这种剧毒人类病原体感染的抗病毒方案。在这里，我们证明，如果预防性给予，磷酸奥司他韦可有效预防食蟹猴的严重疾病。由于奥司他韦耐药突变体的出现，治疗方案的疗效降低，导致四只动物之一死亡。 这强调了在流行病或大流行早期实施联合治疗和疫苗接种策略的重要性。 （Feldmann 等人，mBio 2019）。 1997年禽流感病毒的爆发凸显了高致病性H5N1病毒有可能导致人类严重疾病。因此，需要针对 H5N1 病毒的有效疫苗来应对全球大流行的潜在威胁。我们生成了基于 VSV 的重组 H5N1 流感病毒载体，以证明该平台用于快速作用的泛 H5 流感病毒疫苗的可行性。我们发现单剂量表达全长血凝素 (HAfl) 的 VSV 载体足以提供 100% 的保护。致死攻击前 3 天施用时，疫苗载体起效迅速，可提供均匀的保护。此外，单次疫苗接种可诱导交叉保护性 H5 特异性抗体，并保护小鼠免受各种 H5 2 分支病毒的致命攻击，凸显了基于 VSV 的 HAfl 作为泛 H5 流感病毒紧急疫苗的潜力。 （3）中东呼吸综合征冠状病毒（MERS-CoV）：我们进行了两个主要的治疗项目，抗体疗效和瑞德西韦治疗。我们测试了单克隆抗体 LCA60 在普通狨猴 MERS-CoV 感染模型中的预防效果。在病毒攻击前一天静脉注射 LCA60 可产生中等的临床治疗益处，包括减少呼吸系统受累。尽管与对照组相比，LCA60治疗的动物的病毒肺负荷没有减少，但肺部的病理变化较少。因此，可以实施预防性 LCA60 治疗，以减轻确诊的 MERS-CoV 患者接触者的疾病负担。此外，我们还测试了瑞德西韦在中东呼吸综合征冠状病毒感染的恒河猴模型中预防和治疗的疗效。接种前24小时开始预防性瑞德西韦治疗完全预防了MERS-CoV引起的临床疾病，强烈抑制了MERS-CoV在呼吸组织中的复制，并防止了肺部病变的形成。接种后 12 小时开始的瑞德西韦治疗也提供了明显的临床益处，临床症状减少，肺部病毒复制减少，肺部病变的存在和严重程度降低。这里提供的数据支持在 MERS 临床试验中测试瑞德西韦治疗的疗效。 （4）严重急性呼吸系统综合症冠状病毒（SARS-CoV-2）：自2020年2月以来，我们主要从事COVID项目。我们建立了叙利亚仓鼠疾病模型，用于发病机制研究和对策开发（Rosenke 等人，手稿正在准备中）。我们已经建立了一个体外药物筛选管道，将使用叙利亚仓鼠模型将有前途的候选药物送入体内测试（Jarvis 等人，Antivir Ther 2020；Rosenke 等人，J Antimicrobial Chemotherap，已提交）。这些研究正在进行中。我们进一步评估了羟氯喹（HCQ）在两种动物疾病模型中的预防/治疗效果。在恒河猴疾病模型中，标准人类疟疾 HCQ 预防和治疗并没有显着改善临床结果，也没有减少上呼吸道和下呼吸道中 SARS-CoV-2 的复制/脱落。同样，当用于预防或治疗时，标准人类疟疾剂量和高剂量的 HCQ 对叙利亚仓鼠疾病模型中的临床疾病或 SARS-CoV-2 动力学（复制/脱落）都没有任何有益作用。这两种临床前动物模型的结果可能有助于指导临床使用 HCQ 预防/治疗 COVID-19。最后，我们快速开发了repRNA-CoV2S，这是一种稳定且具有高免疫原性的候选疫苗，由采用新型脂质无机纳米颗粒（LION）配制而成的RNA复制子组成，旨在增强疫苗的稳定性、递送和免疫原性。我们已经证明，肌肉注射 LION/repRNA-CoV2S 会引发强烈的抗 SARS-CoV-2 刺突蛋白 IgG 抗体同种型，表明小鼠体内出现 1 型 T 辅助反应以及有效的 T 细胞反应。重要的是，在非人灵长类动物中仅进行初免给药可引发有效中和 SARS-CoV-2 的抗体反应以及表明 1 型 T 辅助反应的 T 细胞反应。这些数据支持进一步开发 LION/repRNA-CoV2S 作为预防 SARS-CoV-2 感染的候选疫苗（Erasmus 等人，Sci Transl Med 2020）。