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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的“不可治疗”的目标，疾病相关的蛋白质-蛋白质相互作用的细胞内，小分子药物无法阻断。特别是抗体，可以被设计成几乎结合任何蛋白质并抑制蛋白质功能，但尚未用于针对这些目标。这主要是因为，在细胞内递送足够量的折叠的功能性抗体方面的挑战。这项提案的目的是是利用蛋白质自组装构建细胞内抗体载体，并评估递送抗体。通过利用六聚体蛋白束（HEX）和通用抗体结合肽在SPAB（SPABs）中，仅含有蛋白质的纳米级复合物可以与抗体形成。我们的假设是 HEX抗体纳米载体将显著增加细胞内抗体摄取抗体，并且将使得治疗性抗体能够结合至胞质靶标。我们的初步数据显示， 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，可以广泛地应用于细胞内。适用于未来的细胞内抗体治疗。为达到这一目标，假设。(1)制备并表征HEX抗体的大小、稳定性和抗体载量纳米载体(2)测量模型抗癌的细胞摄取、胞质定位和治疗功能通过HEX纳米载体递送的抗体。(3)确定HEX纳米载体的治疗潜力，加入抗HER 2靶向抗体，将载体导向乳腺癌细胞，并评估血液 HEX纳米载体在体内的循环时间和免疫原性。预计会有两个主要成果。一是将产生创新蛋白质纳米载体，其促进功能性治疗剂的胞质递送抗体的它将为细胞内抗体治疗提供概念证明，以抑制蛋白质，包括与靶向抗体的组合。其次，这项工作将测量必要的关键属性，翻译、血液循环时间和免疫原性。这将为细胞内抗体与目前成功治疗人类疾病的细胞外抗体相当的工程。