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: 10587723
• 负责人: Samuel Lai
• 金额: $ 68.84万
• 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/10587723
• 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; Epithelial; 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; Pharmacology; 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; base; crosslink; effective therapy; efficacy testing; improved; in vitro Model; in vivo; in vivo Model; molecular targeted therapies; neutralizing monoclonal antibodies; novel; parainfluenza virus; pathogen; 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.

人类副流感病毒（PIV）和偏肺病毒（MPV）是下呼吸道感染的常见原因 婴儿和幼儿的呼吸道感染以及免疫受损者呼吸道疾病的主要原因 成人和老年人。不幸的是，这两种病毒都没有治疗方法或疫苗，只能提供支持性治疗 提供医疗服务。有趣的是，PIV 和 MPV 有许多共同的病理和临床表现： 呼吸道合胞病毒（RSV）。事实上，目前已知 PIV 和 MPV 均不会导致血液中的病毒血症。 感染患者，表明这两种感染都严格局限于呼吸道，与传播后代的 RSV 类似 病毒仅来自受感染细胞的顶端表面。这种独特的病理生理学意味着后代 病毒在传播到邻近细胞之前必须穿过气道粘液（AM）。这反过来又促使我们 开发一种病原体特异性抗病毒药物，可以从物理上限制呼吸道内感染的传播。 我们最近在粘液中发现了一种新的抗体效应器功能——将单个病原体捕获在粘液中 基于 IgG-Fc 和粘蛋白之间精心调整的亲和力 – 并开发了一个增强 mAb 的平台 作用于粘膜表面。我们假设递送至气道的“粘膜捕获”单克隆抗体可以直接 通过拦截和捕获 AM 中脱落的后代病毒来干预病毒生命周期，快速消灭 通过自然粘膜纤毛清除作用捕获气道中的病毒，并实现体内有效治疗。在 为了支持这一策略，我们设计了可有效捕获 AM 中 RSV 的 mAb，并表明雾化 从感染后第 3 天开始，向感染 RSV 的新生羔羊有效输送“粘液捕获”单克隆抗体 3天内将肺组织中的感染性病毒载量降低至不可检测的水平，其中病毒RNA 与载体对照相比，支气管组织减少了 11 倍。这些结果激励我们去探索 类似的方法是否可以有效治疗 PIV 和 MPV 感染。为了支持这个申请， 我们设计了对两种病毒的不同毒株具有皮摩尔亲和力的抗体，并证明 我们可以有效地捕获人类AM中的病毒，并限制高分化人类体外感染的传播 气道上皮（WD-HAE）在气液界面生长。 在本提案中，我们将继续利用哺乳动物和酵母展示进行亲和力成熟的工作 产生可广泛结合并中和 PIV 和 MPV 不同毒株的高亲和力 mAb（目标 1）。我们将 验证这些 mAb 是否可以捕获新鲜未稀释的人 AM 中的 PIV 和 MPV，以及是否可以抑制 WD-HAE 培养物中预先确定的 PIV 和 MPV 感染的传播（目标 2）。然后我们将推进 两种病毒的先导单克隆抗体，用于在仓鼠鼻部感染模型中进行评估（目标 3）。通过启用增强 mAb 在粘液分泌物中发挥作用，我们希望能够帮助为改进的分子靶向治疗铺平道路 针对所有主要粘膜表面的广谱病原体的治疗和预防，提供 这是解决当前呼吸道感染药物干预措施空白的有力选择。