Engineered “muco-trapping” antibodies for inhaled therapy of parainfluenza and human metapneumovirus infections

用于副流感和人类偏肺病毒感染吸入治疗的工程化“粘膜捕获”抗体

• 批准号: 10707403
• 负责人: Samuel Lai
• 金额: $ 68.83万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-19 至 2027-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10707403
• 关键词: Address; Adult; Affinity; Air; Antibodies; Apical; Back; Binding; Biological Assay; Blood; Bronchiolitis; Caring; Cells; Child; Clinical; Cytopathology; Diffuse; Dose; Ebola; Elderly; Engineering; Epithelium; Evaluation; Functional disorder; Goals; Hamsters; Herpesvirus 1; Histology; Hour; Human; Human Metapneumovirus; Immune; Immunocompromised Host; Immunoglobulin G; Immunotherapy; In Vitro; Individual; Infant; Infection; Inflammation; Influenza; Inhalation; Inhalation Therapy; Intercept; Intervention; Lead; Libraries; Life Cycle Stages; Liquid substance; Lower Respiratory Tract Infection; Lung; Medical; Metapneumovirus; Methods; Modeling; Monoclonal Antibodies; Mucins; Mucociliary Clearance; Mucous Membrane; Mucous body substance; Mus; Nebulizer; Neonatal; Neutrophil Infiltration; Nose; Parainfluenza; Particulate; Pathologic; Pathology; Patients; Penetration; Pneumonia; Polysaccharides; Proliferating; Property; Prophylactic treatment; Proteins; Recombinants; Research Personnel; Respiratory Syncytial Virus Infections; Respiratory Tract Infections; Respiratory syncytial virus; Structure of parenchyma of lung; Surface; Tissues; Vaccines; Vagina; Viral; Viral Load result; Viral load measurement; Viremia; Virion; Virus; Virus Shedding; Work; Yeasts; airway epithelium; antibody engineering; antigen binding; asthma exacerbation; crosslink; effective therapy; efficacy testing; improved; in vitro Model; in vivo; in vivo Model; molecular targeted therapies; neutralizing monoclonal antibodies; novel; parainfluenza virus; pathogen; pharmacologic; purge; respiratory; respiratory virus; transmission process; viral RNA

Human parainfluenza virus (PIV) and metapneumovirus (MPV) are common causes of lower respiratory tract infections in infants and young children, and major causes of respiratory illness in immune compromised adults and the elderly. Unfortunately, there are no therapy or vaccine for either viruses, and only supportive medical care is available. Interestingly, PIV and MPV share many pathological and clinical manifestations as Respiratory Syncytial Virus (RSV). Indeed, neither PIV nor MPV are known to cause viremia in the blood of infected patients, indicating both infections are strictly localized in the airways similar to RSV that shed progeny viruses exclusively from the apical surface of infected cells. Such unique pathophysiology implies progeny viruses must traverse airway mucus (AM) before spreading to neighboring cells. This in turn motivated us to develop a pathogen-specific antiviral that could physically limit the spread of the infections within the airways. We recently discovered a novel Ab effector function in mucus – trapping individual pathogens in mucus based on carefully-tuned affinity between IgG-Fc and mucins – and developed a platform for enhancing mAb function at mucosal surfaces. We hypothesize that “muco-trapping” mAb delivered to the airways can directly intervene with the viral life cycle by intercepting and trapping shed progeny viruses in AM, rapidly eliminate trapped viruses from the airways by natural mucociliary clearance, and enable effective therapy in vivo. In support of this strategy, we engineered mAbs that potently trap RSV in AM, and showed that nebulized delivery of “muco-trapping” mAb to RSV-infected neonatal lambs beginning on Day 3 post infection effectively reduced the infectious viral load in lung tissues to non-detectible levels within 3 days, with viral RNA in bronchial tissues reduced by 11-fold compared to vehicle control. These results motivated us to explore whether a similar approach may be effective in treating PIV and MPV infections. In support of this application, we have engineered antibodies with picomolar affinity to diverse strains of both viruses, and demonstrated that we could effectively trap viruses in human AM and limit spread of infection in vitro in well-differentiated human airway epithelium (WD-HAE) grown at the air-liquid interface. In this proposal, we will continue our work with affinity maturation using mammalian and yeast display to produce high affinity mAb that broadly bind and neutralize diverse strains of PIV and MPV (Aim 1). We will validate whether these mAbs can trap PIV and MPV in fresh undiluted human AM, and whether they can inhibit the spread of pre-established PIV and MPV infections in WD-HAE cultures (Aim 2). We will then advance the lead mAb for both viruses for evaluation in a hamster nasal infection model (Aim 3). By enabling enhanced mAb function in mucus secretions, we expect we will help pave the way for improved, molecularly-targeted therapies and prophylaxis against a broad spectrum of pathogens across all major mucosal surfaces, providing a powerful option addressing the current gap in pharmacological interventions for respiratory infections.

人副帕弗鲁氏病毒（PIV）和Metapneumovirus（MPV）是下呼吸道的常见原因 婴儿和幼儿的道感染，以及免疫系统中呼吸道疾病的主要原因 成人和较早的人。不幸的是，任何一种病毒都没有治疗或疫苗，只有支持 可以提供医疗服务。有趣的是，PIV和MPV共享许多病理和临床表现 呼吸综合病毒（RSV）。确实，众所周知，PIV和MPV在 受感染的患者，表明这两种感染都严格定位在气道中，类似于脱落后代的RSV 仅来自感染细胞的顶端表面的病毒。如此独特的病理生理学意味着进步 病毒在扩散到相邻细胞之前必须穿越气道粘液（AM）。这反过来促使我们 开发一种病原体特异性抗病毒，可以物理限制气道内感染的扩散。 我们最近在粘液中发现了一种新颖的AB效应功能 - 诱捕粘液中的个体病原体 基于IgG-FC和粘蛋白之间经过精心调整的亲和力 - 并开发了一个增强mAb的平台 粘膜表面的功能。我们假设传递到气道的“粘液捕获” mAb可以直接 通过拦截和捕获AM中的后代病毒来介入病毒生命周期，迅速消除 通过天然粘膜扫除率从气道中捕获病毒，并在体内启用有效的治疗。在 支持这种策略，我们设计了可能将RSV捕获在AM中的MAB，并表明这是雾化的 将“粘液捕获” mAb传递给RSV感染的新生儿羔羊，从3天开始感染后第3天 在3天内将肺组织中的传染性病毒负荷降低至不可检测水平，病毒RNA在 与媒介物对照相比，支气管组织减少了11倍。这些结果促使我们探索 类似方法是否可以有效治疗PIV和MPV感染。为了支持此应用程序， 我们已经对两种病毒的潜水菌株具有皮摩尔亲和力的设计抗体，并证明 我们可以有效地将病毒捕获在人类AM中，并在体外限制感染良好的人的体外传播 气道上皮（WD-HAE）在空气界面生长。 在此提案中，我们将使用哺乳动物和酵母显示 产生高亲和力mAb，可以广泛结合和中和PIV和MPV的潜水员菌株（AIM 1）。我们将 验证这些mAb是否可以将PIV和MPV捕获在新鲜的单人类AM中，以及它们是否可以抑制 在WD-HAE培养物中预先建立的PIV和MPV感染的传播（AIM 2）。然后，我们将推进 在仓鼠鼻感染模型中，这两种病毒的铅mab（AIM 3）。通过增强 Mucus分泌物中的MAB功能，我们希望我们将为改进，分子靶向的方法铺平道路 针对所有主要粘膜表面各种病原体的疗法和预防 解决呼吸道感染药物干预措施中当前差距的强大选择。