Admin-Core-001

管理核心-001

• 批准号: 10794918
• 负责人: Richard Thomas Wyatt
• 金额: $ 37.25万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-02-04 至 2026-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10794918
• 关键词: Admin; Core; 001

The Administrative Core A, led by Dr. Richard Wyatt, will administer support for Projects 1 and 2 and Cores B and C, under the umbrella of this new HIVRAD proposal. This Core will coordinate an administrative partnership plan to maintain good communications within the HIVRAD working group and relevant external researchers, advisors and stakeholders related to design of novel cleavage-independent trimers and the analysis of B cell responses generated by vaccination of these HIV-derived envelope glycoprotein (Env) timers into non-human primates (NHPs). The Administrative Core will establish a HIVRAD website as well as cloud data storage accessible to all component PIs or/and designates. A major scientific objective, to elicit and define tier 2 cross-neutralizing B cell and antibody responses (bNAbs) in NHPs following vaccination of novel Env trimers, will be forwarded by Core A. The purpose of Core A is embodied by the successful competition of the following tasks. We will perform project management, while implementing a developed plan to ensure the success of the overall P01 program. This approach will be accomplished by the Program Director, enhanced by real-world learning processes in the accomplished previous Wyatt.Scripps HIVRAD. In Project 2, the Core A will foster studies to better define the expressed NHP heavy and light chain repertoire to determine lineages and the levels of somatic hypermutation elicited by heterologous trimer prime:boost immunization in NHPs. We will examine responses elicited by the novel uncleaved near-native NFL trimers in functional and structural detail to identify neutralizing Abs for genetic analysis. We will use NGS analysis to follow Ab lineages to define affinity maturation, as well as to investigate if the Abs are archived in long-lived compartments (bone marrow). We will apply deep sequencing computational analysis of vaccine-induced Env-specific B cell responses to define populations of epitope-specific Abs in NHPs. We will investigate the evolution of Env-specific Ab lineages following immunization using the NGS-based tracing module within IgDiscover. We will define the maturation pathways of mAbs and use this information to re-design trimer immunogens to drive the responses in broadly neutralizing directions. We will use structural analysis of Env-specific serum Abs (Core C), and isolated mAbs from vaccinated NHPs (Project 2) for iterative trimer redesign. The HIVRAD research team will leverage our previous finding to advances the field’s understanding of vaccine-elicited B cell responses to neutralizing determinants of HIV-1 in small animals and NHPs to a level not previously approachable. The continued development of novel B cell sorting probes (ordered cleavage-independent, Avi-tagged NFL trimers), advances in cloning methods, Ab expression, B cell germinal center and lymph node analytical capacities, Ab germline gene identification and Ab lineage tracing through NGS make this an extremely timely and important application. In this new application we will continue “move the bar” forward towards the elicitation of bAbs by subunit Env vaccination using novel, covalent particulate display as well as the exciting recent advances in lipid nanoparticle mRNA expression of cell-surface tethered NFL trimers.

由Richard Wyatt博士领导的行政核心A将管理对项目1和2以及核心B的支持 和C，在这个新的HIVRAD提案的保护伞下。该核心将协调行政伙伴关系 计划在HIVRAD工作组和相关外部研究人员之间保持良好的沟通， 与新型非切割依赖性三聚体设计和B细胞分析相关的顾问和利益相关者 通过将这些HIV衍生的包膜糖蛋白（Env）计时体接种到非人 灵长类动物（NHP）。行政核心将建立一个HIVRAD网站以及云数据存储 可访问所有组件PI或/和指定。一个主要的科学目的是在接种新型Env三聚体后，在NHP中引发和定义2级交叉中和B细胞和抗体应答（bNAb）， 由Core A转发。核心A的目的通过以下任务的成功竞争来体现。 我们将执行项目管理，同时实施制定的计划，以确保整体的成功 P01程序。这种方法将由项目总监完成，并通过现实世界的学习得到加强 在完成之前的Wyatt.Scripps HIVRAD。在项目2中，核心A将促进研究， 更好地定义表达的NHP重链和轻链库，以确定谱系和体细胞表达水平， 由异源三聚体引发的超突变：NHP中的加强免疫。我们将检查响应 由新的未切割的近天然NFL三聚体在功能和结构细节上引起，以鉴定中和 做基因分析。我们将使用NGS分析来跟踪Ab谱系以定义亲和力成熟，以及 研究Ab是否在长寿命隔室（骨髓）中存档。我们将应用深度测序 疫苗诱导的Env特异性B细胞应答的计算分析， NHP中的Abs。我们将研究Env特异性Ab谱系在免疫后的进化，使用免疫后的Env特异性Ab谱系。 IgDiscover中基于NGS的跟踪模块。我们将定义单克隆抗体的成熟途径， 信息来重新设计三聚体免疫原，以在广泛中和的方向上驱动应答。我们将使用 Env特异性血清Ab（核心C）和来自接种疫苗的NHP的分离mAb（项目2）的结构分析， 迭代三聚体重新设计。HIVRAD研究小组将利用我们以前的发现来推进该领域的研究。 了解疫苗诱导的B细胞对小动物中HIV-1中和决定簇的反应， NHP达到以前无法达到的水平。新型B细胞分选探针的持续开发（已订购 不依赖于切割，Avi标记的NFL三聚体），克隆方法的进展，Ab表达，B细胞增殖 中心和淋巴结分析能力，通过NGS进行Ab生殖系基因鉴定和Ab谱系追踪 这是一个非常及时和重要的应用程序。在这个新的应用程序中，我们将继续“移动酒吧” 使用新的共价颗粒展示，通过亚单位Env疫苗接种， 以及令人兴奋的最新进展，脂质纳米颗粒mRNA表达的细胞表面拴系NFL三聚体。