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Identification of Lamprey Antibodies Capable of Noninvasive Brain Drug Delivery

能够无创脑部药物输送的七鳃鳗抗体的鉴定

基本信息

批准号：
10186832
负责人：
ERIC V SHUSTA
金额：
$ 32.92万
依托单位：
UNIVERSITY OF WISCONSIN-MADISON
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-06-15 至 2023-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10186832
关键词：
Acquired Immunodeficiency Syndrome Affinity Alzheimer&apos s Disease American Antibodies Antibody Specificity Antibody-drug conjugates Antigen Receptors Binding Biodistribution Biological Products Blood Blood - brain barrier anatomy Blood Circulation Brain Brain-Derived Neurotrophic Factor Carbohydrates Cell membrane Cerebrum Characteristics Clinical Complementary DNA Coupling Data Development Diffuse Disease model Dose Drug Delivery Systems Drug Kinetics Drug Transport Endothelium Exhibits Family Fc Receptor Gene Proteins Generations Genes Goals Hagfish Human Immune Immune system Immunization Immunize Immunoglobulin G Immunoprecipitation Insulin Lampreys Lead Leucine-Rich Repeat Libraries Link Liposomes Low Density Lipoprotein Receptor Low-Density Lipoproteins Lymphocyte Malignant neoplasm of brain Mammals Mediating Medicine Mining Mus Oryctolagus cuniculus Parkinson Disease Patients Pharmaceutical Preparations Pharmacologic Substance Pharmacology Pharmacotherapy Polysaccharides Preparation Proteins Reagent Research Route Side Specificity Stains Stroke Structure Surface System Techniques Technology Therapeutic Transferrin Translating Translations Validation Vertebrates Yeasts base blood-brain barrier permeabilization brain tissue cerebral microvasculature dalton drug candidate drug development high-throughput drug screening immunogenic improved in vivo innovation intravenous administration intravenous injection lipophilicity member nanoparticle nervous system disorder novel novel therapeutics pharmacokinetic model programs receptor response screening small molecule small molecule therapeutics stroke model tandem mass spectrometry targeted delivery therapeutic DNA trafficking transcytosis uptake vector

项目摘要

ABSTRACT Millions of people worldwide suffer from neurological diseases such as Alzheimer's disease, stroke, and brain cancer. Advances in protein/gene profiling techniques and high throughput drug screening technologies have spawned many new drug candidates. However, the blood-brain barrier (BBB) has impeded the development and clinical realization of this new generation of neurotherapeutics by restricting the brain uptake of most small molecule therapeutics, and prohibiting brain uptake of protein- and gene-based medicines. A promising noninvasive brain delivery strategy takes advantage of endogenous BBB transport mechanisms as a means to shuttle drug cargo from the blood to the brain. Such receptor-mediated transport systems can be targeted using the exquisite specificity of antibodies that are in turn linked to a drug payload that can include small molecules, proteins, or DNA therapeutics. After binding to the receptor on the blood side, the antibody- drug conjugate acts as an artificial substrate for the transporter and is transcytosed from the blood, across the BBB, and into the brain. Current approaches have yielded limited brain uptake because the targeted transporters are ubiquitously expressed, and the antibody targeting reagents have a low BBB permeability. Therefore, this proposal is focused on the identification and validation of novel delivery vectors and their cognate BBB transporters that can mediate improved transport efficiency. Rather than deploying traditional mammalian antibody technology, we describe a new strategy that employs lamprey antibodies known as Variable Lymphocyte Receptors (VLRs) to target the BBB. Lampreys and humans last shared a common ancestor >500 million years ago, and due to this tremendous evolutionary divergence, even highly conserved mammalian proteins and carbohydrates are immunogenic in lampreys. By leveraging these unique aspects of the lamprey immune system with innovative screening technologies, we anticipate that the proposed research will provide new BBB-targeting VLRs capable of trafficking into the brain. To achieve these goals, lampreys were immunized with mouse brain microvessel plasma membrane preparations, and staining with the resultant polyclonal antiserum demonstrated that VLRs clearly recognize the in vivo BBB and bind to multiple unique glycan structures. The lymphocyte cDNA of immunized lampreys was then used to create a yeast display library consisting of millions of VLRs that will be screened using an innovative screening approach to select BBB-binding and trafficking monoclonal VLRs. These brain-targeting VLRs will be validated both by pharmacokinetic profiling and their capacity to elicit a pharmacologically- relevant response in a murine stroke model. Finally, the BBB-targeting VLRs will be employed to identify the cognate BBB transporter and any associated glycoforms. Those VLRs exhibiting significant and specific brain uptake would represent new, noninvasive brain drug delivery vectors that could be powerful in the treatment of debilitating neurological disease.

抽象的全世界有数百万人患有阿尔茨海默病、中风等神经系统疾病脑癌。蛋白质/基因分析技术和高通量药物筛选技术的进展催生了许多新的候选药物。然而，血脑屏障（BBB）阻碍了通过限制大脑摄取来开发和临床实现新一代神经治疗药物大多数小分子疗法，并禁止大脑摄取基于蛋白质和基因的药物。一个有前途的非侵入性脑输送策略利用内源性 BBB 运输机制作为意味着将药物从血液运送到大脑。这种受体介导的转运系统可以是使用抗体的精确特异性进行靶向，这些抗体又与药物有效负载相关联，其中可以包括小分子、蛋白质或 DNA 疗法。抗体与血液侧的受体结合后，药物缀合物作为转运蛋白的人工底物，从血液中转胞吞，穿过 BBB，进入大脑。目前的方法对大脑的吸收有限，因为目标转运蛋白普遍表达，抗体靶向试剂的血脑屏障通透性较低。因此，该提案的重点是新型递送载体的识别和验证以及它们的同源 BBB 转运蛋白可以提高运输效率。而不是部署传统的哺乳动物抗体技术，我们描述了一种采用七鳃鳗抗体的新策略称为可变淋巴细胞受体 (VLR)，以 BBB 为目标。七鳃鳗和人类最后共享一个超过5亿年前的共同祖先，由于这种巨大的进化分歧，甚至高度七鳃鳗中保守的哺乳动物蛋白质和碳水化合物具有免疫原性。通过利用这些独特的通过创新的筛选技术，我们预计七鳃鳗免疫系统的各个方面拟议的研究将提供能够运输到大脑中的新的针对 BBB 的 VLR。为了实现这些目标，用小鼠脑微血管质膜对七鳃鳗进行了免疫接种制剂，并用所得的多克隆抗血清染色证明 VLR 可以清楚地识别体内 BBB 并结合多个独特的聚糖结构。免疫七鳃鳗淋巴细胞cDNA 然后用于创建由数百万个 VLR 组成的酵母展示文库，这些 VLR 将使用选择 BBB 结合和运输单克隆 VLR 的创新筛选方法。这些针对大脑的 VLR 将通过药代动力学分析及其引发药理学的能力进行验证小鼠中风模型中的相关反应。最后，以 BBB 为目标的 VLR 将用于识别同源 BBB 转运蛋白和任何相关的糖型。那些 VLR 表现出重要且特定的大脑摄取将代表新的、非侵入性脑部药物输送载体，可以有效治疗以下疾病：使人衰弱的神经系统疾病。

项目成果

期刊论文数量（8）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Antibody-Targeted Liposomes for Enhanced Targeting of the Blood-Brain Barrier.

DOI：
10.1007/s11095-022-03186-1
发表时间：
2022-07
期刊：
PHARMACEUTICAL RESEARCH
影响因子：
3.7
作者：
Ye, Zhou;Gastfriend, Benjamin D.;Umlauf, Benjamin J.;Lynn, David M.;Shusta, Eric, V
通讯作者：
Shusta, Eric, V

Yeast display biopanning identifies human antibodies targeting glioblastoma stem-like cells.