Novel Single Domain Antibodies with Multivalency and Multispecificity

具有多价性和多特异性的新型单域抗体

基本信息

批准号：
8399006
负责人：
Rihe Liu
金额：
$ 28.74万
依托单位：
UNIV OF NORTH CAROLINA CHAPEL HILL
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-12-09 至 2016-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8399006
关键词：
Address Affinity Amino Acid Sequence Amino Acids Animal Model Animals Antibodies Apoptosis Avidity Bacteria Binding Biological Products Breast Calmodulin Cell Proliferation Chemicals Collaborations Colorectal Complex Cytoplasm Development Diagnosis Diagnostic Disadvantaged Drug Delivery Systems ERBB2 gene ERBB3 gene Engineering Epidermal Growth Factor Receptor Epitopes Evolution Extracellular Domain Family Family member Gene Delivery Generations Genes Head and Neck Cancer Human Image In Vitro Interdisciplinary Study Investigation Lead Length Libraries Ligands Lung Malignant Neoplasms Malignant neoplasm of pancreas Messenger RNA Methods Mono-S Nanotechnology Neoplasm Metastasis Pancreatic Ductal Adenocarcinoma Patients Penetration Peptide Sequence Determination Peptides Plant Leaves Play Post-Translational Protein Processing Proteins Receptor Protein-Tyrosine Kinases Regulation Research Role SHFM1 gene Signal Transduction Site Small Interfering RNA Specificity Structure Surgical Oncologist System Technology Tertiary Protein Structure Therapeutic Agents Time Tissues Xenograft procedure base calcium phosphate cancer cell cancer diagnosis cancer therapy cancer type cell motility cost cost effective directed evolution humanized monoclonal antibodies immunogenicity in vivo interest malignant stomach neoplasm member mouse model nanoparticle neoplastic cell next generation novel receptor scaffold self assembly success targeted delivery tumor tumorigenesis

项目摘要

DESCRIPTION (provided by applicant): ErbB/HER receptors play essential roles in propagating signals that regulate cell proliferation, differentiation, motility, and apoptosis. Significantly, aberrant regulation of the ErbB/HER family of receptor tyrosine kinases and their ligands is a common occurrence in many human tumors. Indeed, EGFR, HER2, and HER3 have been diagnostically and therapeutically validated as targets in a wide variety of human malignancies such as lung, colorectal, breast, head and neck, gastric and pancreatic cancers. Therefore, novel protein biopharmaceuticals that comprehensively target ErbB/HER family members could have significant applications in targeted diagnosis and therapy of different types of cancer. This project addresses a critical need for developing cost-effective targeting ligands that recognize and bind HER family members with desired avidity and multispecificity. The project is directed at using a powerful in vitro protein selection method called mRNA display to evolve single domain antibodies (SDAs) with human origin that tightly and specifically bind to the extracellular domain of EFRG, HER2 or HER3, respectively, from a human SDA domain library with an unusually high diversity. The resulting monomeric SDAs will be used as the basis for the generation of heterodimeric SDAs that bind to a HER member of interest at two nonoverlapping epitopes with synergistic avidity effect. In addition, we will develop an efficient and universal self-assembly system that allows for reversible and controllable loading of HER-binding SDAs to nanoparticles to acquire desired multispecificity. The resulting SDAs would have a number of advantages, including high target-binding affinity, ease to achieve desired avidity, multivalency and multispecificity, minimal immunogenicity due to human origin, small size for better tissue penetration, in addition to significantly reduced manufacturing costs due to high expression levels in bacteria. The simplicity and stability of these SDAs also greatly facilitate their conjugation with various nanoparticles for translational applications. The resulting SDAs will be examined in vivo in pancreatic ductal adenocarcinoma (PDAC) xenograft animal models by targeted delivery of nanoparticles containing siRNAs against several metastasis signature genes that are important for PDAC tumorigenesis and metastasis. The success of the project will lead to the development of cost-effective, targeted biopharmaceuticals for cancer diagnosis and therapy.

描述（由申请人提供）：ErbB/HER 受体在调节细胞增殖、分化、运动和凋亡的信号传播中发挥重要作用。值得注意的是，受体酪氨酸激酶 ErbB/HER 家族及其配体的异常调节在许多人类肿瘤中很常见。事实上，EGFR、HER2 和 HER3 已被诊断和治疗验证为多种人类恶性肿瘤的靶标，例如肺癌、结直肠癌、乳腺癌、头颈癌、胃癌和胰腺癌。因此，全面靶向ErbB/HER家族成员的新型蛋白质生物药物可能在不同类型癌症的靶向诊断和治疗中具有重要应用。该项目解决了开发具有成本效益的靶向配体的迫切需求，这些配体能够以所需的亲合力和多特异性识别并结合 HER 家族成员。该项目旨在使用一种称为 mRNA 展示的强大体外蛋白质选择方法，从人类 SDA 结构域进化出人类来源的单域抗体 (SDA)，分别紧密且特异性地结合 EFRG、HER2 或 HER3 的胞外结构域图书馆具有异常高的多样性。由此产生的单体 SDA 将用作生成异二聚体 SDA 的基础，异二聚体 SDA 与感兴趣的 HER 成员在两个不重叠的表位处结合，具有协同亲合力效应。此外，我们将开发一种高效且通用的自组装系统，允许将 HER 结合 SDA 可逆且可控地装载到纳米颗粒上，以获得所需的多特异性。由此产生的 SDA 将具有许多优点，包括高靶点结合亲和力、易于实现所需的亲和力、多价性和多特异性、由于人类来源而具有最小的免疫原性、尺寸小以实现更好的组织渗透，此外由于在细菌中高表达。这些 SDA 的简单性和稳定性也极大地促进了它们与各种纳米粒子的缀合以用于转化应用。由此产生的 SDA 将在胰腺导管腺癌 (PDAC) 异种移植动物模型中进行体内检查，方法是靶向递送含有 siRNA 的纳米颗粒，针对几种对 PDAC 肿瘤发生和转移很重要的转移特征基因。该项目的成功将有助于开发用于癌症诊断和治疗的具有成本效益的靶向生物制药。