Scientific Core: Structural Proteomics

科学核心：结构蛋白质组学

• 批准号: 10725051
• 负责人: Andrew Barrett Ward
• 金额: $ 18.1万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-06-01 至 2028-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10725051
• 关键词: Affinity; Animal Model; Animals; Antibodies; Antibody Binding Sites; Antibody Response; Antigens; Apical; B cell repertoire; B-Lymphocytes; B-cell receptor repertoire sequencing; Cells; Clinical Research; Complementarity Determining Regions; Cryoelectron Microscopy; Data; Development; Electron Microscopy; Epitopes; Genes; HIV; HIV Antibodies; HIV vaccine; Human; Immune Sera; Immune response; Immunize; Immunoglobulin Somatic Hypermutation; Individual; Knock-in; Knock-in Mouse; Maps; Methods; Molecular; Monoclonal Antibodies; Mus; Negative Staining; Polysaccharides; Proteomics; Resolution; Specificity; Structure; Subunit Vaccines; Techniques; Vaccine Design; Vaccines; Work; animal data; data format; database query; design; iterative design; mouse model; multiple omics; neutralizing antibody; nonhuman primate; pathogen; phase I trial; polyclonal antibody; pre-clinical; single-cell RNA sequencing; vaccine development; vaccine evaluation

Project Summary/Abstract Rational, structure-based immunogen design for HIV vaccine development has begun to deliver promising results in both, pre-clinical non-human primate (NHP) and clinical studies such as the IAVI G001 phase I trial [NCT03547245]. Moreover, advances in single cell RNAseq and electron microscopy-based polyclonal antibody mapping (EMPEM) have enabled analysis of both vaccine and pathogen-induced antibody responses at unprecedented resolution, further enabling rational vaccine design. EMPEM and single cell B cell analysis deliver fundamentally different data formats that if properly integrated, can illuminate immune responses at unprecedented detail. We recently extended the original negative stain EMPEM method, capable of distinguishing polyclonal antibody epitope specificities and angle of approach, to high resolution cryoEM-based EMPEM to yield molecular details of epitope-paratope interfaces. In the best cases we can derive some sequence information for the complementarity determining regions (CDRs) from structural data, identify Vh/Vl gene usage, and query databases of antibody BCR sequences to directly generate monoclonal antibodies. As such, we can short cut the laborious efforts to generate epitope specific monoclonal antibodies using conventional methods. These molecular details are critical for identifying how bona fide human germline bnAb precursors engage germline targeting immunogens such as the V2 apex targeting Q23 immunogen. EMPEM is also critical for evaluating polyclonal antibody responses in BCR knock-in mice that have been immunized with germline targeting immunogens that are designed to prime and mature antibodies with specific features in CDRs that resemble known broadly neutralizing antibodies (bnAbs). The Structural Proteomics Core will be highly integrated into all both Project 1 and 2 as well as the Animals and Data Cores within this P01 application, providing unique structural data to evaluate, refine, and optimize multi-epitope germline targeting immunogens for development as candidate HIV vaccines.

项目摘要/摘要 艾滋病毒疫苗开发的理性，基于结构的免疫原设计已开始提供有希望的 结果是临床前非人类灵长类动物（NHP）和临床研究，例如IAVI G001 I期试验 [NCT03547245]。此外，单细胞RNASEQ和基于电子显微镜的多克隆抗体的进展 映射（EMPEM）可以分析疫苗和病原体诱导的抗体反应 空前的分辨率，进一步实现了理性疫苗设计。 EMPEM和单细胞B细胞分析传递 根本不同的数据格式，如果正确整合，可以照亮免疫反应 前所未有的细节。我们最近扩展了原始的负污渍empem方法，能够 区分多克隆抗体表位特异性和接近角度，与基于冷冻的高分辨率 empem产生表位 - 参数界面的分子细节。在最好的情况下，我们可以得出一些 互补性确定区域（CDR）的序列信息，确定VH/VL 基因用法和抗体BCR序列的查询数据库直接生成单克隆抗体。作为 这样，我们可以缩短使用艰苦的努力，以生成特定表位的单克隆抗体 常规方法。这些分子细节对于确定真正的人类种系BNAB至关重要 前体接合靶向免疫原的种系，例如靶向Q23免疫原的V2顶点。 empem是 对于评估已对BCR敲击小鼠的多克隆抗体反应至关重要的 生殖线靶向免疫原，这些免疫原旨在具有CDR中具有特定特征的抗体和成熟抗体 该抗体（BNAB）类似于已知的广泛中和抗体。结构蛋白质组学核心将高度 集成到所有项目1和2中，以及该P01应用程序中的动物和数据核心， 提供独特的结构数据来评估，完善和优化靶向免疫原的多诊断 作为候选HIV疫苗的发育。