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感染，基于抗体的疫苗必须能够持续地引起 广泛中和抗体（bnAbs）。bnAb诱导已被证明在人类和非人类中是困难的。 灵长类动物（NHP），部分原因是bnAb需要复杂的发育途径才能成熟， 来自对HIV-1包膜糖蛋白（Env）的脱靶应答的竞争，以及罕见的前体B细胞 频率.在这个项目中，我们将开发候选免疫原，以解决这些问题，通过 各种方法。首先，我们将使用基于结构和哺乳动物展示技术， 增强我们的主要候选物Q23.17 MD 39的生殖系靶向能力。使用这些方法，我们 将开发能够靶向V2-apex和CD 4结合位点bnAb表位的天然样三聚体。这些 三聚体免疫原将另外被工程化以通过最小化竞争来限制脱靶应答 来自高度免疫显性的非中和抗体谱系。此外，我们将使用这些结构 开发新的“生殖细胞靶向SHIV”，并通过SHIV感染来识别增强免疫原 可以引导广度的发展。此外，通过对Env. 抗体共同进化，我们的目标是确定在感染的灵长类动物中bnAb发展的决定因素，这将进一步 告诉我们免疫原的设计过程最后，我们将这些基因佐剂复杂的发展， 项目1和项目3的平台进展，以及我们的新型DNA发射纳米颗粒（DLNP） 平台我们已经证明DLNP改善了免疫原性，同时不需要复杂的cGMP 这阻碍了传统纳米颗粒疫苗的临床开发步伐。然后我们将 使用多种具有人类B细胞谱系的创新小鼠模型向下选择免疫原， 重现人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进入GMP生产提供基准。我们将在一个 NHP模型的异源挑战，并通过这些组合的方法，我们的目标是证明， 首次在NHP中保护免受异源2级挑战。