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.

项目总结/摘要 用于HIV疫苗开发的基于结构的合理免疫原设计已经开始提供有希望的 在临床前非人灵长类动物（NHP）和临床研究（如IAVI G 001 I期试验）中均获得了结果 [NCT 03547245]。此外，单细胞RNAseq和基于电子显微镜的多克隆抗体的进展 EMPEM（EMPEM）已经能够分析疫苗和病原体诱导的抗体反应， 前所未有的分辨率，进一步实现合理的疫苗设计。EMPEM和单细胞B细胞分析提供 根本不同的数据格式，如果适当整合，可以阐明免疫反应， 前所未有的细节。我们最近扩展了原来的负染色EMPEM方法，能够 区分多克隆抗体表位特异性和接近角度，以高分辨率cryoEM为基础 EMPEM产生表位-互补位界面的分子细节。在最好的情况下，我们可以得到一些 从结构数据中获得互补决定区（CDR）的序列信息，鉴定Vh/Vl 基因使用，查询抗体BCR序列数据库，直接生成单克隆抗体。作为 因此，我们可以缩短产生表位特异性单克隆抗体的艰苦努力， 常规方法。这些分子细节对于鉴定真正的人类生殖系bnAb 前体与生殖系靶向免疫原如V2顶点靶向Q23免疫原接合。EMPEM是 这对于评估BCR基因敲入小鼠中的多克隆抗体应答也是至关重要的， 生殖系靶向免疫原，其被设计用于引发和成熟在CDR中具有特异性特征的抗体 类似于已知的广泛中和抗体（bnAb）。结构蛋白质组学核心将高度 集成到所有项目1和2以及本P01应用程序中的动物和数据核心中， 提供独特的结构数据以评估、改进和优化多表位种系靶向免疫原 作为HIV疫苗的候选人。