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 C，在新的 HIVRAD 提案的框架下，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， 与新型切割独立三聚体设计和 B 细胞分析相关的顾问和利益相关者 将这些 HIV 衍生的包膜糖蛋白 (Env) 计时器接种到非人类体内所产生的反应 管理核心将建立一个 HIVRAD 网站以及云数据存储。 所有组成部分 PI 或/和指定人员均可获得一个主要科学目标，即在接种新型 Env 三聚体后引发和定义 NHP 中的 2 级交叉中和 B 细胞和抗体反应 (bNAb)。 由Core A转发。Core A的目的通过以下任务的成功竞争来体现。 我们将进行项目管理，同时实施制定的计划，以确保整体的成功 P01 项目将由项目总监完成，并通过现实世界的学习得到加强。 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水平 由异源三聚体引发的超突变：NHP 中的加强免疫接种我们将检查反应。 由新型未切割的近天然 NFL 三聚体在功能和结构细节上引起，以识别中和 我们将使用 NGS 分析来追踪 Ab 谱系，以定义亲和力成熟度以及亲和力成熟度。 为了调查抗体是否存档在长寿区室（骨髓）中，我们将应用深度测序。 对疫苗诱导的 Env 特异性 B 细胞反应进行计算分析，以定义表位特异性群体 我们将使用 NHP 来研究免疫后 Env 特异性抗体谱系的进化。 IgDiscover 中基于 NGS 的追踪模块我们将定义 mAb 的成熟途径并使用它。 我们将使用重新设计三聚体免疫原以驱动广泛中和方向的反应的信息。 Env 特异性血清抗体（核心 C）的结构分析，以及从细菌 NHP 中分离出的单克隆抗体（项目 2） HIVRAD 研究团队将利用我们之前的发现来推进该领域的迭代三聚体重新设计。 了解疫苗引发的 B 细胞对小动物中 HIV-1 中和决定因素的反应，以及 NHP 达到以前无法达到的水平 持续开发新型 B 细胞分选探针（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 细胞生发 中心和淋巴结分析能力、抗体种系基因鉴定和通过 NGS 进行抗体谱系追踪 使其成为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”。 使用新型共价颗粒显示，通过亚基 Env 疫苗接种来诱导 bAb 以及细胞表面束缚的 NFL 三聚体的脂质纳米颗粒 mRNA 表达方面令人兴奋的最新进展。