Refinement of DNA-Launched NanoParticles decorated with Apex and CD4bs B cell lineage targeting Envs (DLNP-ACEs)

用 Apex 和 CD4bs B 细胞谱系靶向 Envs (DLNP-ACE) 修饰的 DNA 启动纳米粒子的精制

• 批准号: 10589588
• 负责人: Daniel Kulp
• 金额: $ 116.34万
• 依托单位: WISTAR INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-12-08 至 2027-11-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10589588
• 关键词: Address; Adjuvant; Adoptive Transfer; Affinity; Animal Model; Animals; Antibodies; Antibody Repertoire; Antigens; Apical; Artificial nanoparticles; B-Lymphocytes; Benchmarking; Binding; Binding Sites; Cell Lineage; Clinic; Complex; Cryoelectron Microscopy; Cyclic GMP; DNA; DNA Structure; Development; Directed Molecular Evolution; Electron Microscopy; Engineering; Epitope Mapping; Epitopes; Formulation; Frequencies; Genes; Genetic; Glycoproteins; Goals; HIV; HIV vaccine; HIV-1; HIV-1 vaccine; Human; Immune; Immunity; Immunization; In Vitro; Individual; Investigation; Knock-in Mouse; Laboratories; Maps; Measures; Methods; Modality; Modeling; Modification; Molecular; Monitor; Mus; Mutation; Pathway interactions; Pattern; Plasma; Primates; Process; Production; Protein Subunits; Protocols documentation; Publications; Recombinants; Recurrence; Reporting; Rhesus; Somatic Mutation; Structure; Structure of germinal center of lymph node; Subunit Vaccines; Surface; Techniques; Testing; Time; Vaccine Design; Vaccines; Variant; Virus; Virus Diseases; acronyms; cell type; clinical development; design; global health; immunogenic; immunogenicity; improved; in vivo; innovation; lead candidate; lead optimization; lead series; mouse model; nanoparticle; neutralizing antibody; nonhuman primate; novel; pre-clinical; programs; prophylactic; response; simian human immunodeficiency virus; synthetic construct; trafficking; vaccine candidate; vaccine strategy; vaccine trial

Project 2 - Project Summary In order to protect against HIV-1 acquisition, antibody-based vaccines must be able to consistently elicit broadly neutralizing antibodies (bnAbs). bnAb elicitation has proven difficult in humans and non-human primates (NHPs), in part because of the complex developmental pathways bnAbs require to mature, competition from off-target responses to the HIV-1 envelope glycoprotein (Env), and rare precursor B cell frequency. In this project we will develop candidate immunogens to address each of these problems through a variety of approaches. First, we will use structure-based and mammalian display techniques to further enhance the germline targeting ability of our lead candidate, Q23.17 MD39. Using these approaches, we will develop native-like trimers capable of targeting the V2-apex and CD4 binding site bnAb epitopes. These trimer immunogens will additionally be engineered to limit off-target responses by minimizing competition from highly immunodominant non-neutralizing antibody lineages. Furthermore, we will use these constructs to develop novel “germline targeting SHIVs”, and through SHIV-infection will identify boosting immunogens that can guide the development of breadth. Additionally, through structural analysis of the process of Env- Ab coevolution, we aim to identify determinants of bnAb development in infected primates, that will further inform our immunogen design process. Finally, we will complex these with genetic adjuvants developed in project 1 and platform advances in project 3, as well as with our novel DNA-Launched NanoParticle (DLNP) platform. We have shown DLNPs improve the immunogenicity while not requiring the complex cGMP processes that hamper the pace of clinical development for traditional nanoparticle vaccines. We will then down-select immunogens using a variety of innovative mouse models that harbor human B cell lineages, recapitulate human B cell competition and have tunable bnAb precursor frequencies. These mice will provide a benchmark for our lead candidates to be advanced into GMP production. We will test them in a NHP model for heterologous challenge, and through these combined approaches, we aim to demonstrate, for the first time, protection from heterologous tier-2 challenge in NHPs.

项目2-项目摘要 为了防止HIV-1采集，基于抗体的疫苗必须能够始终如一地引起 广泛中和抗体（BNAB）。事实证明，BNAB启发很难在人类和非人类中 Primes（NHP），部分原因是BNAB需要成熟的复杂发展途径， 竞争来自靶向脱靶反应到HIV-1信封糖蛋白（ENV）和稀有前体B细胞的竞争 频率。在这个项目中，我们将开发候选免疫原，以通过 各种方法。首先，我们将使用基于结构和哺乳动物的显示技术来进一步 增强我们的铅候选者的种系靶向能力，Q23.17 MD39。使用这些方法，我们 将开发能够靶向V2-APEX和CD4结合位点BNAB表位的天然三聚体。这些 Trimer Immunogens还将通过最小化竞争来限制脱离目标的响应 来自高度免疫主导的非中和抗体谱系。此外，我们将使用这些结构 为了开发新颖的“生殖线靶向湿因”，并通过Shiv感染将确定增强免疫原分 这可以指导广度的发展。此外，通过结构分析环境的过程 进化，我们旨在确定受感染私人中BNAB发展的决定者，这将进一步 通知我们的免疫原设计过程。最后，我们将使这些与遗传调节器复杂化 项目1和平台进展3，以及我们的新型DNA启动纳米颗粒（DLNP） 平台。我们已经显示DLNP提高了免疫原性，而不需要复杂的CGMP 妨碍传统纳米颗粒疫苗的临床发育速度的过程。然后我们会 使用各种具有人类B细胞谱系的创新小鼠模型的下选择免疫原， 概括人B细胞竞争，并具有可调的BNAB前体频率。这些老鼠会 为我们的主要候选人提供基准，以便将其推进GMP生产。我们将在 NHP用于异源挑战的模型，通过这些联合方法，我们旨在证明， 首次保护NHP中的异源2挑战。