Cleaved, stabilized HIV-1 Env trimers for structural and vaccine studies

 描述(申请人提供)：项目1的目标是基于一系列HIV-1基因设计和评估新版本的SOSIP gp140三聚体，并结合旨在改善其结构、抗原性和免疫原性的各种序列修改。项目1中设计的三聚体将由核心A生产，提供给项目2进行结构研究，评估项目1内的抗原性和其他性质，还将发送到多个外部 其他研究的合作者，包括动物的免疫原性实验(项目1也将参与分析)。因此，项目1将密切参与整个HIVRAD的科学计划。HIVRAD的PI约翰·P·摩尔博士将执导总部设在纽约威尔康奈尔医学院(WCMC)的项目1。摩尔博士将与罗吉尔·W·桑德斯博士密切合作，后者将领导阿姆斯特丹学术医学中心的一个小组。项目1的具体目标是：目标1：改进目前最先进的SOSIP三聚体BG505 SOSIP.664的设计，用于结构和免疫原性研究。我们将利用高分辨率X射线和冷冻-EM结构和动物免疫原性研究的信息设计新的SOSIP三聚体。我们将为bNAbs创建针对种系祖先的变异三聚体，并基于婴儿-BG505内的序列进化创建序列三聚体。其他目标是减少三聚体的异质性，从而提高结晶潜力，掩盖可能作为免疫诱骗的非NAB表位，更好地将bNAb表位呈现给免疫系统，并推动抗体进化向bNAbs的发展。目的2：从不同遗传亚型中鉴定出与BG505 SOSIP.664特性相似的多个SOSIP三聚体。我们将结合我们积累的三聚体设计经验、我们对基于BG505的三聚体(A亚型)性质的知识以及生物信息学方法，根据其他亚型，特别是B和C亚型来识别和/或设计新的变种。目标是创造一种配方，使我们能够从许多/大多数基因型制造高质量的SOSIP三聚体，从而扩大两个项目中可用于疫苗和基于结构的研究的三聚体的范围。目的3：设计并帮助评估使用SOSIP三聚体的免疫原性研究。HIVRAD团队已经与包括但不限于CHAVI-ID和IAVI在内的几个组织建立了合作，为动物免疫研究提供基于三聚体的免疫原，作为人类临床试验的一部分。项目1团队将在HIVRAD内部指导委员会的参与下，与相关合作者共同设计具体的实验，并将通过进行中和和相关血清学分析来为研究分析做出贡献。 相关性：这种多部位HIVRAD应用的总体目标是帮助开发一种预防性HIV-1疫苗，其基础是通过合理设计、结构相关的包膜糖蛋白三聚体诱导病毒中和抗体。核心A将在项目1和2中设计SOSIP三聚体，并将其提供给项目团队和外部合作者。申请的目标与公共卫生和人类福利高度相关。

 DESCRIPTION (provided by applicant): The goals of Project 1 are to design and evaluate new versions of SOSIP gp140 trimers based on a range of HIV- 1 genes and incorporating various sequence modifications intended to improve their structural, antigenicity and immunogenicity properties. The trimers designed in Project 1, with essential structure-based input from Project 2, will be produced by Core A, provided to Project 2 for structural studies, assessed for antigenicity and other properties within Project 1, and also sent to multiple external collaborators for additional research, including immunogenicity experiments in animals (which Project 1 will also be involved in analyzing). Project 1 will, therefore, be intimately involved inthe scientific program of the entire HIVRAD. Dr. John P. Moore, the PI of the HIVRAD, will direct Project 1, which will be based at the Weill Cornell Medical College (WCMC), New York. Dr. Moore will work closely with Dr. Rogier W. Sanders, who will lead a component at the Academic Medical Center, Amsterdam. The Specific Aims of Project 1 are: Aim 1: To improve the design of the currently most advanced SOSIP trimers, BG505 SOSIP.664, for structural and immunogenicity studies. We will design new SOSIP trimers using information from high-resolution x-ray and cryo-EM structures and animal immunogenicity studies. We will create variant trimers that target germline ancestors for bNAbs, and sequential trimers based on sequence evolution within infant-BG505. Other goals are to reduce trimer heterogeneity and thereby improve crystallization potential, mask non-NAb epitopes that may act as immunological decoys, better present bNAb epitopes to the immune system, and drive Ab evolution toward the development of bNAbs. Aim 2: To identify multiple SOSIP trimers from different genetic subtypes with properties comparable to BG505 SOSIP.664. We will combine our accumulated experience of trimer design, our knowledge of the properties of the BG505-based trimers (subtype A), and a bioinformatics approach, to identify and/or design new variants based on other subtypes, particularly B and C. The goals are to create a recipe that will enable us to make high quality SOSIP trimers from many/most genotypes and thereby expand the range of trimers available for vaccine- and structure-based studies in both Projects. Aim 3: To design and help evaluate immunogenicity studies using SOSIP trimers. The HIVRAD team has established collaborations with several groups, including but not limited to CHAVI-ID and IAVI, to provide trimer-based immunogens for animal immunization studies as part of the pathway towards human clinical trials. The Project 1 team will jointly design specific experiments with the relevan collaborators, with the involvement of the HIVRAD's Internal Steering Committee, and will contribute to the analysis of the studies by performing neutralization and related serology assays. RELEVANCE: The overall objective of this multi-site HIVRAD application is to help develop a preventative HIV-1 vaccine based on the induction of virus neutralizing antibodies by rationally designed, structurally relevant envelope glycoprotein trimers. Core A will make the SOSIP trimers designed in Projects 1 and 2, and provide them to both project teams and external collaborators. The goals of the application are highly relevant to public health and human welfare.