Bifunctional antibodies with targeted CNS delivery against West Nile Virus

具有针对西尼罗河病毒的靶向中枢神经系统递送的双功能抗体

• 批准号: 8839547
• 负责人: Qiang Chen
• 金额: $ 46.04万
• 依托单位: ARIZONA STATE UNIVERSITY-TEMPE CAMPUS
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-06-01 至 2017-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8839547
• 关键词: Address; Adverse effects; Affect; Affinity; Agrobacterium; Antibodies; Antibody-Dependent Enhancement; Binding; Biomass; Bispecific Antibodies; Blood - brain barrier anatomy; Brain; C57BL/6 Mouse; Cessation of life; Characteristics; Chinese Hamster Ovary Cell; Communicable Diseases; Complement 1q; Complement-Dependent Cytotoxicity; Conscious; Cyclic GMP; Data; Development; Disease; Dose; E protein; Elderly; Encephalitis; Endocytosis; Endothelial Cells; Engineering; Ensure; Epitopes; Fc Receptor; Future; Generations; Genetic Engineering; Grant; Hamsters; Human; Immunocompromised Host; In Vitro; Infection; Inflammation; Inflammatory; Intravenous; Licensing; Link; Meningitis; Modeling; Mus; Neuraxis; Pattern; Peripheral; Permeability; Phase; Plant Proteins; Plants; Polysaccharides; Process; Production; Proteins; Rattus; Reagent; Regulation; Risk; Rodent; Route; Safety; Survival Rate; Technology; Testing; Therapeutic; Transferrin Receptor; Transgenic Organisms; Variant; Vertebrates; Virus Diseases; West Nile virus; adverse outcome; base; clinical care; combat; cost; cost effective; env Gene Products; glycosylation; human INSR protein; improved; intraperitoneal; large scale production; mouse model; neoplastic; nervous system disorder; novel therapeutics; receptor; scale up; theories; transcytosis; vector

PROJECT SUMMARY/ABSTRACT West Nile Virus (WNV) causes infection in the central nervous system (CNS) in several vertebrate animal species. Humans infected with WNV can develop meningitis and encephalitis, and the elderly and immunocompromised are at greatest risk for severe neurological disease and death. New threats of WNV globally and the lack of available treatments warrant studies to develop effective therapeutics and production technologies that can rapidly transfer candidate therapies into the clinical care setting in a cost- conscious manner. We recently developed a plant-derived humanized MAb with promising therapeutic potential, with a desired human N-linked glycosylation pattern. This MAb (hE16) binds to a highly conserved epitope on the envelope protein of virtually all isolates worldwide and shows promising post-exposure therapeutic activity. Nonetheless, our studies show that peripheral delivery of hE16 has a limited window of efficacy in rodents: administration of a single dose of hE16 through an intravenous or intraperitoneal route at day 5 post infection or earlier improves survival rates. In comparison, delivery of hE16 directly into the brain at day 6 after infection improved protection against lethal WNV infection in hamsters. Our preliminary data show that a genetically engineered bifunctional MAb variant of hE16 (TfR-Bif) with the potential for enhanced crossing of the blood-brain barrier (BBB) can be expressed in CHO cells or plants. TfR-Bif retains its ability to bind and neutralize WNV, but importantly gains the ability to bind transferrin receptor (TfR) and endocytose into mouse brain endothelial cells. In the R21 phase of this grant, we will use the mouse TfR- specific TfR-Bif as a proof-of-principle in a mouse model of WNV infection. We will test the hypothesis that TfR-Bif can achieve higher levels in the CNS and extends the window of treatment against WNV encephalitis. To address critical safety issues in the brain, we will also identify TfR-Bif glycoforms that offer full bifunctionality with limited or specific Fc receptor (Fc¿R) or C1q binding to eliminate potential antibody- dependent enhancement (ADE) of virus infection or pathogenic inflammation. In the R33 phase, we will develop an analogous bifunctional mAb with human therapeutic potential and test if plants provide distinctive advantages in safety, production cost, and scalability for large-scale production under cGMP. In addition to generating novel therapeutic reagents for WNV, this collaborative study will provides a platform for delivery of MAbs to the CNS, which should be applicable to the treatment of other infectious, inflammatory, or neoplastic diseases in the CNS.

项目概要/摘要 西尼罗河病毒 (WNV) 导致多种脊椎动物的中枢神经系统 (CNS) 感染 物种。感染西尼罗河病毒的人类可患上脑膜炎和脑炎，老年人和 免疫功能低下的人患严重神经系统疾病和死亡的风险最大。西尼罗河病毒的新威胁 在全球范围内，由于缺乏可用的治疗方法，需要研究开发有效的治疗方法和 生产技术可以以成本快速地将候选疗法转移到临床护理环境中 有意识的方式。我们最近开发了一种植物源性人源化单克隆抗体，具有良好的治疗前景 潜力，具有所需的人类 N 连接糖基化模式。该 MAb (hE16) 与高度保守的 全世界几乎所有分离株的包膜蛋白上都有表位，并显示出有希望的暴露后效果 治疗活动。尽管如此，我们的研究表明 hE16 的外周递送窗口有限 对啮齿类动物的功效：通过静脉内或腹膜内途径施用单剂量的 hE16 感染后第 5 天或更早进行注射可提高存活率。相比之下，将 hE16 直接递送至 感染后第 6 天，大脑增强了对仓鼠致命性西尼罗河病毒感染的保护作用。我们的初步 数据显示，hE16 的基因工程双功能 MAb 变体 (TfR-Bif) 具有潜力 增强的血脑屏障 (BBB) 穿越能力可以在 CHO 细胞或植物中表达。 TfR-Bif 保留 它具有结合和中和 WNV 的能力，但重要的是获得了结合转铁蛋白受体 (TfR) 的能力， 内吞入小鼠脑内皮细胞。在本次资助的 R21 阶段，我们将使用小鼠 TfR- 特异性 TfR-Bif 作为 WNV 感染小鼠模型的原理验证。我们将检验以下假设： TfR-Bif 可以在 CNS 中达到更高的水平，并延长针对 WNV 的治疗窗口 脑炎。为了解决大脑中的关键安全问题，我们还将鉴定可提供以下功能的 TfR-Bif 糖型： 具有有限或特定 Fc 受体 (Fc¿R) 或 C1q 结合的完整双功能，以消除潜在的抗体- 病毒感染或致病性炎症的依赖性增强（ADE）。在R33阶段，我们将 开发一种具有人类治疗潜力的类似双功能单克隆抗体，并测试植物是否提供 在安全性、生产成本以及cGMP下大规模生产的可扩展性方面具有独特的优势。在 除了为西尼罗河病毒生产新型治疗试剂外，这项合作研究还将提供一个平台 用于将单克隆抗体输送至中枢神经系统，这应适用于治疗其他感染性疾病， 中枢神经系统炎症或肿瘤性疾病。