Rationalizing glycoengineering strategies for immunotherapeutic antibodies

免疫治疗抗体糖工程策略的合理化

基本信息

批准号：
10377400
负责人：
ERIC JOHN SUNDBERG
金额：
$ 47.12万
依托单位：
EMORY UNIVERSITY
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：
2020-04-08 至 2025-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10377400
关键词：
Active Sites Address Affect Alanine Animal Model Antibodies Autoantibodies Autoimmune Diseases Autoimmunity Bacteria Binding Carbohydrates Chemistry Clinical Complement Complex Coupled Cryoelectron Microscopy Custom Data Development Endoglycosidases Engineering Enzymes Exhibits Family Fc Receptor Future Glycobiology Glycoengineering Glycoproteins Glycoside Hydrolases Health Homologous Gene Human Hybrids IgG1 Immune response Immune system Immunity Immunoglobulin G Immunologic Receptors Immunotherapeutic agent Inflammatory Knowledge Lead Link Malignant Neoplasms Mannose Molecular Pathologic Processes Pharmaceutical Preparations Physiological Processes Polysaccharides Post-Translational Protein Processing Preparation Property Proteins Reaction Reagent Reproducibility Resolution Role Scanning Site Source Space Perception Specificity Streptococcus pyogenes Structure Substrate Specificity Tertiary Protein Structure Therapeutic Therapeutic Monoclonal Antibodies Therapeutic antibodies Variant X-Ray Crystallography antibody engineering arm base chemical synthesis clinical efficacy design experimental study falls glycosylation human disease human pathogen immunoreaction immunoregulation insight interest next generation novel novel therapeutics receptor therapeutic protein tool

项目摘要

In order to evade host immunity, many bacteria secrete immunomodulatory enzymes. Streptococcus pyogenes, one of the most common human pathogens, secretes unique endoglycosidases, including EndoS, which removes complex-type glycans in a highly specific manner from human IgG antibodies, and its homolog EndoS2, which can additionally remove IgG-linked high-mannose glycans. This renders antibodies incapable of eliciting host effector functions through either complement or Fc γ receptors (FcγRs), providing the bacteria with a survival advantage. Because antibodies are central players in many human immune responses and bridge the innate and adaptive arms of immunity, the analysis and manipulation of the enzymatic activities of EndoS and EndoS2 impact diverse fields in biomedicine. In particular, modifying antibody glycan structures can have significant impacts on their abilities to bind to FcγRs and the subsequent immune system reactions that they induce. The next generation of therapeutic antibodies is already being constructed with modified glycan chemistries to tailor their immune reactions and to increase their clinical potency. EndoS and EndoS2, as antibody-specific glycosidases, and glycosynthases derived thereof, are key enzymes in the future of antibody engineering. We propose that if the molecular mechanisms by which diverse endoglycosidases specifically recognize and hydrolyze distinct glycoprotein substrates are better understood that EndoS and EndoS2 variants can be rationally engineered to create a new class of antibody-modifying enzymes endowed with unique glycan specificities in order to modify antibodies that exhibit enhanced clinical properties. In this proposal, we will address three Specific Aims: (1) to determine the molecular basis of glycan specificity by endoglycosidases; (2) to define the role of carbohydrate binding modules – non-enzymatic protein domains with glycan binding properties – in endoglycosidase specificity and activity; and (3) to elucidate the molecular basis of protein specificity by endoglycosidases. Progress towards these complementary, yet independent, Specific Aims will significantly advance our understanding of glycan-modifying enzymes. Leveraging this knowledge in the context of EndoS and EndoS2 will enhance our ability to customize antibodies, further unleashing their vast therapeutic utility and expanding their positive impact on human health.

为了逃避宿主免疫，许多细菌分泌免疫调节酶链球菌。化脓性杆菌是最常见的人类病原体之一，分泌独特的糖苷内切酶，包括 EndoS、它以高度特异性的方式从人 IgG 抗体及其同源物中去除复合型聚糖 EndoS2，它还可以去除 IgG 连接的高甘露糖聚糖，这使得抗体无法使用。通过补体或 Fc γ 受体 (FcγR) 引发宿主效应功能，为细菌提供因为抗体是许多人类免疫反应的核心参与者连接先天免疫和适应性免疫的桥梁，分析和操纵酶活性 EndoS 和 EndoS2 影响生物医学的各个领域，特别是修饰抗体聚糖结构。可以对其结合 FcγR 的能力以及随后的免疫系统反应产生重大影响他们诱导的下一代治疗抗体已经被构建。聚糖化学来调整其免疫反应并提高其临床效力。作为抗体特异性糖苷酶及其衍生的糖合酶，是未来的关键酶我们提出了不同糖苷内切酶的分子机制。特异性识别和水解不同的糖蛋白底物可以更好地理解 EndoS 和 EndoS2 变体可以合理设计以创建一类新的抗体修饰酶具有独特的聚糖特异性，以修饰表现出增强的临床特性的抗体。提案中，我们将解决三个具体目标：（1）通过以下方式确定聚糖特异性的分子基础： (2) 定义碳水化合物结合模块的作用——非酶蛋白结构域具有聚糖结合特性——糖苷内切酶的特异性和活性；(3) 阐明分子糖苷内切酶的蛋白质特异性的基础。这些互补但独立的进展。利用这一点，具体目标将显着增进我们对聚糖修饰酶的理解。 EndoS 和 EndoS2 背景下的知识将增强我们定制抗体的能力，进一步释放其巨大的治疗效用并扩大其对人类健康的积极影响。