Uncleaved prefusion-optimized trimers on nanoparticles as HIV vaccines

纳米颗粒上未切割的预融合优化三聚体作为 HIV 疫苗

• 批准号: 10307527
• 负责人: Jiang Zhu
• 金额: $ 96.91万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-12-01 至 2023-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10307527
• 关键词: Animal Model; Animals; Antibodies; Antibody titer measurement; Antigens; Artificial nanoparticles; Autologous; B cell repertoire; B-Lymphocytes; Binding; Biological Assay; Clinical Research; Crystallization; Crystallography; Data; Databases; Development; Disease; Electron Microscopy; Engineering; Enzyme-Linked Immunosorbent Assay; Epitope Mapping; Epitopes; Evaluation; Ferritin; Formulation; Future; Generations; Glycoproteins; Goals; HIV Envelope Protein gp120; HIV vaccine; HIV-1; Human; Immune response; Immunization; Immunoglobulin G; In Vitro; Inbred BALB C Mice; Knock-in Mouse; Methods; Monitor; Mus; Mutation; N-terminal; Negative Staining; Oryctolagus cuniculus; Particulate; Primates; Production; Protocols documentation; Reporting; Research; Resolution; Serology test; Serum; Signal Transduction; Source; Structure; Subunit Vaccines; Surface Antigens; T cell response; T-Lymphocyte; Testing; Vaccine Design; Vaccine Research; Vaccines; Virus; Virus-like particle; base; design; glycoprotein structure; immunogenicity; improved; in vivo; in vivo evaluation; nanoparticle; nanoparticulate; neutralizing antibody; next generation sequencing; nonhuman primate; research clinical testing; response; scale up; vaccine candidate; vaccine development; vaccine platform; vaccine strategy

Project Summary The envelope glycoprotein (Env) of human immunodeficiency virus type 1 (HIV-1) harbors the epitopes of broadly neutralizing antibodies (bNAbs) and is the main target of HIV-1 vaccine development. High-resolution structures of BG505 SOSIP.664 trimers have provided a rational basis for vaccine design. Although current gp140 designs can produce native-like trimers for selected strains after optimization, the underlying causes of Env metastability are still unknown and a general design platform applicable to all HIV-1 strains and subtypes remains an unmet challenge. Furthermore, despite the increasing appreciation for the advantages of virus-like particles (VLPs) in bNAb elicitation, nanoparticulate display of native-like trimers has not been rigorously explored in HIV-1 vaccine development. In this R01, we will develop HIV-1 vaccine candidates by combining trimer design, crystallography, electron microscopy (EM), nanoparticle display, immunization, serological assays, antibody isolation, repertoire sequencing, and antibody lineage analysis. In Aim 1, we will investigate the fundamental causes of HIV-1 Env metastability and develop the uncleaved, prefusion-optimized (UFO) trimer platform. We hypothesize that gp41, which contains a structurally strained heptad repeat 1 (HR1) with a disordered N-terminal bend (N-HR1), is the source of metastability. We have designed 1st-generation UFO trimers with a modified N-HR1 and solved their structures. We will develop an optimized UFO platform applicable to diverse Envs with high purity, yield, and stability and select representative UFO trimers for vaccine development. We will assess trimer antigenicity using a panel of bNAbs and non-NAbs and determine their atomic structures. In Aim 2, we will engineer stable gp140 trimer-presenting nanoparticles as vaccine candidates. We hypothesize that self-assembling nanoparticles with underlying 3-fold axes can be used to display native-like gp140 trimers. We have reported gp140 nanoparticles based on the 24-mer ferritin (12 nm) and 60-mer E2p (23nm). We will optimize UFO gp140 nanoparticles based on ferritin, E2p, and a hyperstable 60-mer, I3-01 (25nm) for in vivo testing in Aim 3. We will validate nanoparticle assembly by EM and evaluate nanoparticle antigenicity using a panel of antibodies. In Aim 3, we will assess the immunogenicity and B-cell response for a subset of optimized UFO trimers and nanoparticles in WT mice, rabbits, and nonhuman primates (NHPs). We hypothesize that UFO trimers can induce tier-2 NAbs in all animal models, while gp140 nanoparticles displaying 8-20 UFO trimers can elicit tier-2 NAbs more effectively than trimers alone. We will first screen UFO trimers and nanoparticles in WT mice and assess serum neutralization using purified IgGs. We will select a small set of immunogens for rabbit immunization and advance the most promising ones to NHPs. In our preliminary study, gp140 nanoparticles induced autologous tier-2 NAbs to BG505.T332N in mice and rabbits within 6-8 weeks, supporting our approach. We will isolate vaccine-elicited tier-2 NAbs for functional and structural studies and perform next-generation sequencing (NGS) to study B-cell repertoires and trace NAb lineage development. Our goal is to identify the most effective vaccine candidates for future clinical studies.

项目摘要 人类免疫缺陷病毒1型（HIV-1）的包膜糖蛋白（ENV）具有广泛的表位 中和抗体（BNAB）是HIV-1疫苗发育的主要靶标。高分辨率结构 BG505 SOSIP.664三聚体为疫苗设计提供了合理的基础。尽管当前的GP140设计 优化后可以为选定菌株产生类似天然的三聚体，这是ENV稳定性的根本原因 仍然未知，并且适用于所有HIV-1菌株和亚型的一般设计平台仍然未满足 挑战。此外，尽管人们对病毒样颗粒（VLP）的优势越来越多。 bnab启发，纳米刻度显示本机样三聚体的显示，在HIV-1疫苗中尚未进行严格探索 发展。在此R01中，我们将通过结合三聚体设计，晶体学，， 电子显微镜（EM），纳米颗粒显示，免疫，血清学测定，抗体隔离，曲目 测序和抗体谱系分析。在AIM 1中，我们将调查HIV-1 Env的基本原因 亚竞争性并发展未泄漏的，预融化的（UFO）三聚体平台。我们假设GP41， 包含结构紧张的七肠重复1（HR1），具有无序的N末端弯曲（N-HR1），是 亚竞争能力的来源。我们已经使用修改后的N-HR1设计了第一代的UFO三聚机 结构。我们将开发一个适用于高纯度，产量和 用于疫苗开发的稳定性和选择代表性的UFO三聚体。我们将使用使用 一组BNAB和非NABS，并确定其原子结构。在AIM 2中，我们将设计稳定的GP140 Trimer Extrimented纳米颗粒作为疫苗候选物。我们假设与 下面的3倍轴可用于显示本机样的GP140夹子。我们报道了GP140纳米颗粒 基于24-mer铁蛋白（12 nm）和60-mer E2P（23nm）。我们将优化基于UFO GP140纳米颗粒 在铁蛋白，E2P和AIM 3中的体内测试的I3-01（25nm）上，i3-01（25nm）。我们将验证纳米颗粒 通过EM组装，并使用一系列抗体评估纳米颗粒抗原性。在AIM 3中，我们将评估 在WT小鼠，兔子， 和非人类灵长类动物（NHP）。我们假设UFO三聚体可以在所有动物模型中诱导二级NAB， 而显示8-20 UFO三聚体的GP140纳米颗粒比单独的三聚体更有效地引起层nabs。 我们将首先筛选WT小鼠中的UFO三聚体和纳米颗粒，并使用纯化评估血清中和 Iggs。我们将选择一小群免疫原以进行兔子免疫，并提高最有前途的免疫原分 到NHP。在我们的初步研究中，gp140纳米颗粒在小鼠中诱导自体层Nabs为BG505.T332N 和兔子在6-8周内支持我们的方法。我们将隔离疫苗吸收的层-2 NABS的功能 以及结构研究并执行下一代测序（NGS），以研究B细胞曲目和跟踪NAB 血统的发展。我们的目标是确定未来临床研究的最有效疫苗。