CHEETAH Center for the Structural Biology of HIV Infection, Restriction, and Viral Dynamics

CHEETAH HIV 感染、限制和病毒动力学结构生物学中心

• 批准号: 10508319
• 负责人: WALTHER H MOTHES
• 金额: $ 166.1万
• 依托单位: UNIVERSITY OF UTAH
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-11 至 2027-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10508319
• 关键词: Animal Model; Animals; Anti-Retroviral Agents; Antiviral Agents; Architecture; Biological; Biological Products; Biological Response Modifier Therapy; Biology; Biology of HIV Infection; Capsid; Cell Line; Cells; Complex; Development; Engineering; Foundations; Gene Expression Profile; Generations; Genes; Goals; HIV; HIV-1; Heterochromatin; Image; In Situ; Infection; Integral Membrane Protein; Knock-in; Knock-in Mouse; Knockout Mice; Medicine; Membrane; Membrane Fusion; Messenger RNA; Methods; Methyltransferase; Molecular; Molecular Structure; Mouse Cell Line; Pathway interactions; Patients; Protein Engineering; Proteins; Research; Resistance; Resolution; SETDB1 gene; SIV; Scaffolding Protein; Shapes; Site; Structure; System; Technology; Testing; Therapeutic; Therapeutic Intervention; Transgenic Mice; Viral; Virus; Virus Diseases; base; cell behavior; cell type; delivery vehicle; design; env Gene Products; flexibility; frontier; gene therapy; humanized mouse; imaging approach; macromolecule; model design; mouse model; nanocage; nanomaterials; nanoparticle; new technology; next generation; nonhuman primate; particle; receptor; recruit; self assembly; structural biology; success; viral rebound; virus envelope

PROJECT SUMMARY Studies in this Project are undertaken with the goal of advancing foundations required to tackle new frontiers in HIV-1 biology and medicine, including the development of cure strategies, broad antiviral therapeutics, and methods for delivery of biologic therapeutics into target cells. To these ends, studies in Project 3, Multiscale Analysis and Modulation of Viral Dynamics, will characterize HIV-1 proviral silencing and reactivation over a wide range of resolution scales, from living animals to high-resolution structural studies, and will develop new methods aimed at protecting animals from enveloped viral infections and designing new biologics delivery systems. Studies in Aim 1 (In Situ Architecture of Virus Reactivation) will examine, across multiple size and resolution scales, viruses and associated cells at sites of HIV/SIV rebound in animal models. Specifically, we will: 1) locate and image reactivating virus and associated cells, and 2) define the transcription profiles of infected and neighboring cells at rebound sites. Studies in Aim 2 (Proviral Silencing and Reactivation) seek a molecular and structural understanding of how HIV-1 silencing is established and maintained. To this end, we will determine structures, interactions, and mechanisms important for latency, with a focus on the H3K9-specific methylase SETDB1, and on HUSH, which localizes on H3K9me3 and collaborates with SETDB1 to promote the spread of H3K9me3 and heterochromatin. Studies in Aim 3 (RetroCHMP3 Blocks to Viral Dissemination) will build on our recent discovery of retroCHMP3 proteins, which are naturally-occurring factors that potently inhibit release of enveloped viruses that use the ESCRT pathway for budding, including HIV-1. Our goals are to optimize retroCHMP3 potency in mouse cells lines by increasing protein stability, expression, and restriction activity, and then create transgenic mice that express the optimal constructs and test for broad resistance to enveloped viruses. Studies in Aim 4 (Virus-Inspired Designed Delivery Systems) will generate new virus-inspired technology platforms for intercellular delivery that are simple, robust, and controllable by taking advantage of: 1) recent developments in computational protein design, 2) our previous successes in harnessing the principles of HIV-1 assembly to develop nanoparticles that can direct their own self-assembly and bud from cells, and 3) our new advances for incorporating transmembrane proteins into those particles.

项目总结 该项目的研究目标是推进应对HIV-1新领域所需的基础 生物学和医学，包括治疗策略的发展、广泛的抗病毒疗法和递送方法 将生物治疗药物注入靶细胞。为此，项目3中的研究，病毒的多尺度分析和调制 动力学，将表征HIV-1前病毒沉默和重新激活在广泛的分辨率范围内，从活着 对动物进行高分辨率的结构研究，并将开发旨在保护动物免受围困的新方法 病毒感染和设计新的生物制品输送系统。 在AIM 1(病毒重新激活的原位体系结构)中的研究将检查多个大小和分辨率尺度， 在动物模型中，HIV/SIV部位的病毒和相关细胞反弹。具体地说，我们将：1)定位和成像 重新激活病毒和相关细胞，以及2)定义受感染细胞和邻近细胞在反弹时的转录图谱 网站。 AIM 2(前病毒沉默和重新激活)的研究试图从分子和结构上了解HIV-1是如何 沉默是建立和保持的。为此，我们将确定重要的结构、相互作用和机制 对于延迟，重点是H3K9特定的甲基酶SETDB1，以及定位于H3K9me3和H3K9me3的HUSH 与SETDB1合作，促进H3K9me3和异染色质的传播。 AIM 3(RetroCHMP3阻断病毒传播)的研究将建立在我们最近发现的逆转录CHMP3蛋白的基础上， 它们是自然产生的因子，可以有效地抑制使用ESCRT途径的包膜病毒的释放 包括HIV-1在内的萌芽状态。我们的目标是通过增加蛋白质来优化小鼠细胞系中反转录CHMP3的效力 稳定性、表达和限制活性，然后创造表达最佳结构的转基因小鼠并测试 对包膜病毒具有广泛的抵抗力。 AIM 4(病毒启发设计的传输系统)的研究将产生新的病毒启发技术平台 通过利用以下优势实现简单、强大且可控的细胞间传递：1) 计算蛋白质设计，2)我们以前利用HIV-1组装原理开发的成功 纳米粒子可以引导自己的自我组装和细胞发芽，以及3)我们在掺入 将蛋白质跨膜进入这些颗粒。