Activity-based discovery and optimization of agonist antibodies

基于活性的激动剂抗体的发现和优化

基本信息

批准号：
10159090
负责人：
John Samuel Schardt
金额：
$ 6.97万
依托单位：
UNIVERSITY OF MICHIGAN AT ANN ARBOR
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-04-01 至 2022-04-15
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10159090
关键词：
Active Sites Address Affect Affinity Agonist Antibodies Autoimmune Diseases Biological Biological Assay Biological Models Biological Process Biomedical Technology Biotechnology Cell Separation Cell physiology Cell surface Cells Clinical Trials Computer Models Detection Development Disease Enzymes Evaluation Event Feedback Foundations Frequencies Genetic Engineering Goals Gold Health Human Human Pathology Immunoglobulin Fragments In Vitro Knowledge Lead Length Libraries Ligands Link Malignant Neoplasms Methods Molecular OX40 Pathologic Pathology Pathway interactions Reporter Research Signal Transduction Sorting - Cell Movement Structure-Activity Relationship Surface System T-Cell Activation T-Cell Receptor T-Lymphocyte Technology Therapeutic Therapeutic antibodies Time Validation Variant advanced disease antibody engineering base beta-Lactamase biophysical properties body system design effective therapy extracellular improved improved functioning in silico in vitro Assay innovation innovative technologies lead candidate next generation sequencing novel novel therapeutics receptor receptor-mediated signaling screening therapeutic candidate transcription factor tumor necrosis factor receptor superfamily member 4 wound healing

项目摘要

PROJECT SUMMARY Agonist antibodies, capable of initiating cellular signaling events through interaction at the cell surface, have emerged as a new paradigm in disease treatment with several promising candidates in clinical trials including T cell activating antibodies. However, the discovery of rare antibodies possessing specific biological functions remains a major biotechnological bottleneck. To address this issue, I seek to employ direct function-based antibody screening methods, an emergent biomedical technology using mammalian intracellular reporter systems for which proof-of-concept has recently been shown for the discovery of agonist antibodies. I believe that this innovative technology has the potential to be revolutionary if it can be generalized. Thus, the goal of the proposed research is to establish new fundamental methods for broad use in function-based antibody screening, which could ultimately lay the foundation for advancing disease treatment. I have recently developed novel model systems that enable robust detection of intracellular signaling via the NF-κB pathway initiated from extracellular activation of the T-cell receptors Ox40 and CD137. These receptors were chosen for their availability of well-defined control agonist antibodies that are invaluable for system validation. I propose to use these model systems to gain new understanding of the fundamental mechanisms and principles involved in function-based antibody screening and employ this knowledge toward the development of new biomedical technology and novel therapeutics via three distinct aims. First, I propose to critically evaluate the relationship between biological activity, antibody display level, receptor display level, and antibody molecular format to identify screening methods that ultimately improve the efficiency of agonist antibody discovery. Second, I propose to develop new Ox40 and CD137 reporter cell systems for simplified and improved function-based antibody discovery via genetic engineering of autocatalytic agonist antibody expression that depends on receptor-specific intracellular signaling in order to improve agonist antibody discovery by both simplifying signal detection and amplifying true positive signals. Thirdly, I seek to identify new Ox40 and CD137 agonist antibodies with improved activity by direct function-based screening. Toward this goal, I will employ computational approaches to inform library design in order to increase the likelihood that library variants effectively engage T cell receptors near the active site. I will employ next-generation sequencing to identify lead candidates, which will then be rigorously evaluated via classic T cell activation assays. Finally, I will critically analyze lead sequences in relationship to agonist activity in order to uncover potential molecular determinants of agonist activity and structure-function relationships. Overall, the proposed research has the potential for broad therapeutic impact given that insufficient cellular signaling is involved in a wide range of pathologies including insufficient wound healing, autoimmune disease and cancer.

项目摘要激动剂抗体，能够通过细胞表面的相互作用引发细胞信号事件，具有作为疾病治疗的新范式出现，在包括T 细胞激活抗体。但是，发现具有特定生物学功能的稀有抗体仍然是主要的生物技术瓶颈。为了解决这个问题，我寻求雇员直接基于功能抗体筛选方法，一种使用哺乳动物细胞内报告基因的新兴生物医学技术最近已经显示了概念验证证明的系统以发现激动剂抗体。我相信如果可以将这种创新技术推广，则具有革命性的潜力。那是拟议的研究是建立新的基本方法，以广泛使用基于功能的抗体筛查最终可能为疾病治疗奠定基础。我最近有开发的新型模型系统，可以通过NF-κB途径强耐用细胞内信号传导从T细胞受体OX40和CD137的细胞外激活引发。选择这些受体它们对系统验证的明确控制激动剂抗体的可用性。我建议使用这些模型系统来获得对所涉及的基本机制和原则的新理解在基于功能的抗体筛查和员工中，这些知识旨在发展新的生物医学技术和新型疗法通过三个不同的目标。首先，我建议批判性评估关系在生物活性，抗体显示水平，接收器显示水平和抗体分子格式之间确定最终提高激动剂抗体发现效率的筛选方法。第二，我为开发新的OX40和CD137报告细胞系统的建议，以简化和改进的基于功能通过自催化激动剂抗体表达的基因工程发现抗体，取决于接收器特异性的细胞内信号传导，以通过简化信号来改善激动剂抗体发现检测并放大真正的积极信号。第三，我试图识别新的OX40和CD137激动剂通过基于直接功能的筛选改善活性的抗体。朝着这个目标，我将采用为图书馆设计提供信息的计算方法，以增加图书馆变体的可能性有效地接合活跃部位附近的T细胞受体。我将使用下一代测序来识别铅候选者将通过经典的T细胞激活测定法对其进行严格评估。最后，我会的批判性地分析与激动剂活性关系中的铅序列，以发现潜在的分子激动剂活动和结构功能关系的决定因素。总体而言，拟议的研究具有鉴于细胞信号不足涉及广泛的范围，可能会产生广泛的治疗影响病理学包括逻辑愈合不足，自身免疫性疾病和癌症。