Engineered Protein Nanocarriers for Intracellular Antibody Delivery

用于细胞内抗体递送的工程蛋白质纳米载体

基本信息

批准号：
9387821
负责人：
Julie Champion
金额：
$ 22.59万
依托单位：
GEORGIA INSTITUTE OF TECHNOLOGY
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：
2017-09-05 至 2019-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9387821
关键词：
Address Affinity Antibodies Apoptosis Binding Biodistribution Blood Circulation Time Breast Cancer Cell Carrier Proteins Cells Charge Complex Cytosol Data Disease ERBB2 gene Engineering Enzymes Fc domain Future Generic Drugs Goals Health Hela Cells Human Immunoglobulin G Incubated Individual Literature Malignant Neoplasms Measurement Measures Modeling Modification Outcome Peptides Pharmaceutical Preparations Property Protein Engineering Proteins Recombinant Proteins Recombinants Research Signal Pathway System Technology Testing Therapeutic Therapeutic antibodies Toxic effect Translations Work antibody engineering base clinical translation driving force extracellular human disease immunogenicity in vivo innovation malignant breast neoplasm nanocarrier nanoparticle nanoscale novel novel therapeutics protein function protein protein interaction protein structure response self assembly small molecule therapeutic protein uptake

项目摘要

Engineered Protein Nanocarriers for Intracellular Antibody Delivery There is a long list of “undruggable” targets, disease related protein-protein interactions inside cells that small molecule drugs cannot block. Antibodies, in particular, can be engineered to bind almost any protein and inhibit protein function but have not yet been used against these targets. This is primarily due to the significant, unmet challenge in delivering sufficient amounts of folded, functional antibodies inside cells. The goal of this proposal is to build an intracellular antibody carrier using protein self-assembly and assess the therapeutic activity of delivered antibodies. By utilizing a hexameric protein bundle (HEX) and generic antibody-binding peptides (SPABs), nanoscale complexes containing only protein can be formed with antibodies. Our hypothesis is that the HEX antibody nanocarrier will significantly increase intracellular antibody uptake compared to soluble antibodies, and will enable therapeutic antibody binding to cytosolic targets. Our preliminary data suggests that Hex-SPAB protein nanocarriers do deliver functional antibody into the cytosol and, therefore, could be broadly applicable to future intracellular antibody therapeutics. Three aims have been set to meet the objective and test the hypothesis. (1) Fabricate and characterize the size, stability and antibody loading of HEX antibody nanocarriers. (2) Measure cellular uptake, cytosolic localization, and therapeutic function of model anti-cancer antibodies delivered by HEX nanocarriers. (3) Determine the therapeutic potential of HEX nanocarriers by adding anti-HER2 targeting antibodies to direct carriers to breast cancer cells, and evaluate the blood circulation time and immunogenicity of HEX nanocarriers in vivo. Two main outcomes are expected. First, an innovative protein nanocarrier will be created that facilitates cytosolic delivery of functional therapeutic antibodies. It will provide proof of concept for intracellular antibody therapeutics to inhibit proteins, including combinations with targeting antibodies. Second, this work will measure key properties necessary for translation, blood circulation time and immunogenicity. This will open the door for intracellular antibody engineering comparable to current extracellular antibodies that are successfully treating human disease.

用于细胞内抗体递送的工程蛋白质纳米载体有一长串“不可成药”的靶点，细胞内与疾病相关的蛋白质-蛋白质相互作用，这些靶点很小特别是分子药物不能阻断，可以设计成结合几乎任何蛋白质并抑制。蛋白质功能，但尚未用于针对这些目标，这主要是由于显着的、未满足的。在细胞内提供足够量的折叠功能性抗体是该提案的目标。是利用蛋白质自组装构建细胞内抗体载体并评估其治疗活性通过利用六聚蛋白束 (HEX) 和通用抗体结合肽来递送抗体。（SPAB），仅含有蛋白质的纳米级复合物可以与抗体形成。与可溶性抗体纳米载体相比，HEX 抗体纳米载体将显着增加细胞内抗体的摄取我们的初步数据表明，这将使治疗性抗体能够与胞质靶标结合。 Hex-SPAB 蛋白纳米载体确实将功能性抗体递送到胞质溶胶中，因此可以广泛应用适用于未来的细胞内抗体疗法已经设定了三个目标来满足目标和测试。 (1) 构建并表征 HEX 抗体的大小、稳定性和抗体负载量。 (2) 测量模型抗癌的细胞摄取、细胞质定位和治疗功能 HEX 纳米载体递送的抗体 (3) 通过以下方式确定 HEX 纳米载体的治疗潜力。添加抗 HER2 靶向抗体，将携带者引导至乳腺癌细胞，并评估血液 HEX 纳米载体在体内的循环时间和免疫原性预计有两个主要结果。将创建创新的蛋白质纳米载体，促进功能性治疗药物的胞质传递它将为抑制蛋白质（包括蛋白质）的细胞内抗体疗法提供概念证明。其次，这项工作将测量必要的关键特性。翻译、血液循环时间和免疫原性这将为细胞内抗体打开大门。工程可与目前成功治疗人类疾病的细胞外抗体相媲美。