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

基本信息

批准号：
8473780
负责人：
Qiang Chen
金额：
$ 22.34万
依托单位：
ARIZONA STATE UNIVERSITY-TEMPE CAMPUS
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-06-01 至 2014-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8473780
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

项目摘要

DESCRIPTION (provided by applicant): 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.

描述（由申请人提供）：西尼罗河病毒（WNV）引起多种脊椎动物中枢神经系统（CNS）感染。感染西尼罗河病毒的人类可能会患上脑膜炎和脑炎，而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。全球范围内西尼罗河病毒的新威胁以及可用治疗方法的缺乏需要研究开发有效的治疗方法和生产技术，以便以成本意识的方式将候选疗法快速转移到临床护理环境中。我们最近开发了一种植物源性人源化 MAb，具有良好的治疗潜力，具有理想的人类 N 连接糖基化模式。这种 MAb (hE16) 与全世界几乎所有分离株的包膜蛋白上高度保守的表位结合，并显示出有前景的暴露后治疗活性。尽管如此，我们的研究表明，hE16 的外周递送在啮齿类动物中的功效窗口有限：单次给药在感染后第 5 天或更早通过静脉内或腹膜内途径注射 hE16 剂量可提高存活率。相比之下，在感染后第 6 天将 hE16 直接输送到大脑中可以改善仓鼠对致命性 WNV 感染的保护。我们的初步数据表明，基因工程化的 hE16 双功能 MAb 变体 (TfR-Bif) 具有增强穿过血脑屏障 (BBB) 的潜力，可以在 CHO 细胞或植物中表达。 TfR-Bif 保留了结合和中和 WNV 的能力，但重要的是获得了结合转铁蛋白受体 (TfR) 和内吞至小鼠脑内皮细胞的能力。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。我们将验证 TfR-Bif 可以在中枢神经系统中达到更高水平并延长西尼罗河病毒脑炎治疗窗口的假设。为了解决大脑中的关键安全问题，我们还将鉴定 TfR-Bif 糖型，这些糖型可提供完整的双功能，与有限或特定的 Fc 受体 (Fc?R) 或 C1q 结合，以消除病毒感染或致病性的潜在抗体依赖性增强 (ADE)炎。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MAb，并测试植物是否在安全性、生产成本和cGMP下大规模生产的可扩展性方面具有独特的优势。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向中枢神经系统输送单克隆抗体的平台，该平台应适用于治疗中枢神经系统中的其他感染、炎症或肿瘤性疾病。