Conjugate nanoparticle platform development for HIV-1 envelope immunogens

HIV-1 包膜免疫原的共轭纳米颗粒平台开发

• 批准号: 10369067
• 负责人: KEVIN O SAUNDERS
• 金额: $ 471.96万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-12-17 至 2026-11-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10369067
• 关键词: 2019-nCoV; Affinity; Amino Acids; Animal Model; Antibodies; Antibody Affinity; Antigens; Avidity; B-Cell Antigen Receptor; B-Lymphocytes; Binding Sites; Biomanufacturing; Cell Line; Cell Lineage; Chimeric Proteins; Clinical Trials; Complex; Consumption; Data; Development; Doctor of Philosophy; Documentation; Electron Microscopy; Ensure; Enzyme Activation; Enzymes; Ferritin; Frequencies; Future; Gene Fusion; Generations; Goals; HIV Antigens; HIV Vaccine Trials Network; HIV vaccine; HIV-1; HIV-1 vaccine; Helicobacter pylori; Human; Immune system; Immunization; Immunologics; Immunology; Industry; Infection; Influenza Hemagglutinin; Investigational New Drug Application; Knock-in Mouse; Lead; Link; Macaca mulatta; Molecular Conformation; Mutation; Nature; Negative Staining; Outcome; Pharmaceutical Preparations; Phase I Clinical Trials; Process; Production; Reaction; Regimen; Series; Serum; Site; Somatic Mutation; Statistical Data Interpretation; Structure; Structure of germinal center of lymph node; Testing; Time; Toxic effect; United States Food and Drug Administration; Vaccine Design; Vaccines; Viral; Virion; activation-induced cytidine deaminase; cross reactivity; design; experience; humanized mouse; immunogenic; immunogenicity; improved; innovation; nanoparticle; neutralizing antibody; nonhuman primate; pandemic disease; phase I trial; process optimization; programs; quality assurance; sortase; trafficking; transpeptidation; vaccination strategy; vaccine evaluation; vaccine platform; vaccine trial

ABSTRACT – OVERALL HIV-1 broadly neutralizing antibodies (bnAbs) are protective in animal models of HIV-1 infection, but are not elicited in humans by current vaccine regimens. To elicit bnAbs, the B cell lineage vaccine design approach aims to administer multiple immunogens in a specific sequence to shepherd bnAb maturation through immunologic roadblocks that typically halt bnAb development. One roadblock we recently identified are somatic mutations that encode key amino acids for antibody function but that are rarely made by the somatic mutation enzyme activation-induced cytidine deaminase. Our central vaccine design hypothesis is that antibodies (Abs) encoding these improbable mutations, will be rare; thus, vaccine immunogens will need to have higher affinity for Abs with these desired amino acid changes than Abs without the amino acid changes in order to select for them. The problem facing this strategy is that the only antigen for HIV-1 bnAbs is HIV-1 envelope (Env), which is poorly immunogenic and for which bnAb precursors generally have low affinity. We and others have found these two obstacles can be overcome by designing Envs with high affinity for bnAb precursors and by multimerizing these Envs on nanoparticles (NPs) to provide avidity and improved antigen trafficking to germinal centers. However, Env trimer NPs can have low expression and present misfolded Env trimers that elicit undesired non-neutralizing Abs. This application is significant because it will establish a cGMP-compliant vaccine platform that rapidly generates higher quality HIV-1 Env trimer NP vaccines without time-consuming iterative immunogen design. This platform uses the sortase A enzyme to site-specifically, covalently-link well- folded HIV-1 Env trimers to intact Helicobacter pylori ferritin NPs. The resultant HIV-1 Env trimer sortase A- conjugated NPs (scNPs) display only well-folded Env trimers, and in preliminary studies, have successfully initiated CD4 binding site bnAb lineages in human bnAb precursor knock-in mice and CD4bs nAbs in rhesus macaques. The scNP platform is universal in nature since it can incorporate diverse viral type I fusion proteins by simply adding a 6-amino acid sortase A tag to their C-terminus. In Specific Aim 1, we will compare the ability of monovalent and bivalent HIV-1 Env scNPs to guide affinity maturation of CD4 binding site bnAbs in humanized mice and rhesus macaques. In Specific Aim 2, we will produce and assemble two CD4 binding site-bnAb- targeting HIV-1 Env trimer scNPs (CH505 TF scNP and a second sequential Env trimer scNP) under cGMP conditions. This program will deliver an optimized cGMP process for making scNPs, two cGMP-produced Env trimer scNPs, and additional ferritin and sortase A components for the manufacture of future immunogens. The CH505 TF Env trimer scNPs will be used in a Phase I trial through the HIV Vaccine Trial Network. Ultimately, the impact of this platform is that it will enable multiple Env trimer scNPs to be made rapidly under cGMP, making it feasible to do iterative testing in clinical trials of complete sequential nanoparticle vaccines that target bnAbs.

摘要 - 总体 HIV-1在HIV-1感染的动物模型中受到了广泛中和抗体（BNAB）的保护，但不是 当前的疫苗方案在人类中引起。为了引起bnabs，B细胞谱系疫苗设计方法的目标 通过特定顺序给予多种免疫原，以通过免疫学为牧羊犬BNAB成熟 通常会阻止BNAB开发的障碍。我们最近确定的一个障碍是躯体突变 该编码关键氨基酸以用于抗体功能，但很少由体细胞突变酶制成 激活诱导的胞苷脱氨酶。我们的中央疫苗设计假设是抗体（ABS） 编码这些不可思议的突变将是罕见的；因此，疫苗免疫原子将需要具有较高的亲和力 对于这些所需的氨基酸变化而不是ABS，而没有氨基酸变化，以选择 他们。这种策略面临的问题是，HIV-1 BNABS的唯一抗原是HIV-1信封（ENV），它 免疫原性较差，而BNAB前体通常具有低亲和力。我们和其他人发现 可以通过设计具有对BNAB前体的高亲和力以及通过 对纳米颗粒（NP）上的这些ENV进行多重化，以提供亲戚并改善对生发的抗原运输 中心。但是，Env Trimer NP可以具有低表达，并表现出引起的错误折叠的ENV三聚体 不需要的非中和腹肌。该应用程序很重要，因为它将建立符合CGMP 疫苗平台迅速生成更高质量的HIV-1 ENV触发NP疫苗，而无需耗时 迭代免疫原设计。该平台使用分子酶A酶来特定于站点，共价链接良好 折叠的HIV-1 ENV三聚体，可完整的幽门螺杆菌NP。由此产生的HIV-1 env三聚体分类酶A- 共轭的NP（SCNP）仅显示折叠式的ENV三聚体，并且在初步研究中已成功 在人BNAB前体敲入小鼠中引发的CD4结合位点BNAB谱系和恒河猴的CD4BS NABS 猕猴。 SCNP平台本质上是普遍的 通过简单地将6-氨基酸排序酶添加到其C末端。在特定目标1中，我们将比较能力 人源化的CD4结合位点的亲和力成熟的单价和二价HIV-1 env SCNP 老鼠和恒河猕猴。在特定的目标2中，我们将产生并组装两个CD4结合位点-BNAB- 针对CGMP下的HIV-1 Env Trigger触发SCNP（CH505 TF SCNP和第二个顺序ENV触发SCNP） 状况。该程序将提供一个优化的CGMP流程，用于制造SCNP，两个CGMP生产的Env Trimer SCNP，以及用于生产未来免疫原的成分的其他铁蛋白和分类酶。这 CH505 TF ENV Trimer SCNP将通过HIV疫苗试验网络在I期试验中使用。最终， 该平台的影响是，它将使多个ENV触发SCNP在CGMP下迅速制造，从而使 在针对BNABS的完整顺序纳米颗粒疫苗的临床试验中进行迭代测试。