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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要人类免疫缺陷病毒 1 型 (HIV-1) 的包膜糖蛋白 (Env) 含有广泛的表位中和抗体（bNAb）是HIV-1疫苗开发的主要目标。高分辨率结构 BG505 SOSIP.664三聚体的研究为疫苗设计提供了合理的基础。虽然目前的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亚稳定性的根本原因仍然未知，适用于所有 HIV-1 毒株和亚型的通用设计平台仍未满足挑战。此外，尽管人们越来越认识到病毒样颗粒（VLP）在 bNAb 诱导、类似天然三聚体的纳米颗粒展示尚未在 HIV-1 疫苗中进行严格探索发展。在这个 R01 中，我们将通过结合三聚体设计、晶体学、电子显微镜 (EM)、纳米粒子显示、免疫、血清学测定、抗体分离、曲目测序和抗体谱系分析。在目标 1 中，我们将调查 HIV-1 Env 的根本原因亚稳定性并开发未切割的预融合优化 (UFO) 三聚体平台。我们假设 gp41，其中包含结构紧张的七肽重复序列 1 (HR1) 和无序的 N 末端弯曲 (N-HR1)，是亚稳态的来源。我们设计了带有改进的 N-HR1 的第一代 UFO 三聚体，并解决了它们的问题结构。我们将开发一个适用于各种环境的优化UFO平台，具有高纯度、高产量和高稳定性并选择有代表性的 UFO 三聚体用于疫苗开发。我们将使用以下方法评估三聚体抗原性一组 bNAb 和非 NAB 并确定它们的原子结构。在目标 2 中，我们将设计稳定的 gp140 三聚体呈递的纳米颗粒作为候选疫苗。我们假设自组装纳米粒子底层 3 倍轴可用于显示类似原生的 gp140 三聚体。我们已经报道了 gp140 纳米颗粒基于 24 聚体铁蛋白 (12 nm) 和 60 聚体 E2p (23 nm)。我们将基于 UFO gp140 纳米颗粒进行优化铁蛋白、E2p 和超稳定 60 聚体 I3-01 (25nm)，用于 Aim 3 中的体内测试。我们将验证纳米颗粒通过 EM 组装并使用一组抗体评估纳米颗粒的抗原性。在目标 3 中，我们将评估优化的 UFO 三聚体和纳米粒子子集在 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 2 级 NAb，而显示 8-20 个 UFO 三聚体的 gp140 纳米颗粒可以比单独的三聚体更有效地引发 2 级 NAb。我们将首先在 WT 小鼠中筛选 UFO 三聚体和纳米颗粒，并使用纯化的 IgG。我们将选择一小组用于兔子免疫的免疫原，并推进最有希望的免疫原给 NHP。在我们的初步研究中，gp140 纳米粒子在小鼠中诱导 BG505.T332N 自体 2 级 NAb 和兔子在 6-8 周内，支持我们的方法。我们将分离疫苗引发的 2 级 NAb，以实现功能性和结构研究并进行下一代测序 (NGS) 以研究 B 细胞库并追踪 NAb 谱系发展。我们的目标是为未来的临床研究确定最有效的候选疫苗。