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: 10663373
• 负责人: WALTHER H MOTHES
• 金额: $ 165.23万
• 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/10663373
• 关键词: Animal Model; Animals; Anti-Retroviral Agents; Antiviral Agents; Architecture; Binding; Binding Sites; Biological; Biological Products; Biological Response Modifier Therapy; Biology; Biology of HIV Infection; Capsid; Cell Line; Cells; Collaborations; 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; Modeling; Molecular; Molecular Structure; Mouse Cell Line; Mus; Pathway interactions; Patients; Protein Engineering; Proteins; Research; Resistance; Resolution; SETDB1 gene; SIV; Scaffolding Protein; Shapes; Site; Structure; System; Testing; Therapeutic; Therapeutic Intervention; Transgenic Mice; Viral; Virus; Virus Diseases; Visualization; cell behavior; cell type; delivery vehicle; design; flexibility; frontier; gene therapy; humanized mouse; imaging approach; macromolecule; mouse model; nanocage; nanomaterials; nanoparticle; new technology; next generation; nonhuman primate; particle; receptor; recruit; self assembly; structural biology; success; technology platform; viral rebound

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.

项目摘要 开展本项目的研究，目的是推进解决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，广泛的抗病毒治疗，以及交付方法 生物治疗剂进入靶细胞。为此，项目3中的研究，病毒的多尺度分析和调节， 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尺度，从生活 动物的高分辨率结构研究，并将开发新的方法，旨在保护动物从包膜 病毒感染和设计新的生物制剂输送系统。 目标1（病毒再活化的原位结构）的研究将在多个尺寸和分辨率尺度上研究， 在动物模型中，HIV/SIV部位的病毒和相关细胞反弹。具体来说，我们将：1）定位和图像 重新激活病毒和相关细胞，以及2）确定感染细胞和邻近细胞在反弹时的转录谱 网站. 目标2（前病毒沉默和再激活）的研究旨在从分子和结构上了解HIV-1如何 沉默被建立和维持。为此，我们将确定重要的结构，相互作用和机制， 对于潜伏期，重点是H3 K9特异性甲基化酶SETDB 1和HUSH，其定位于H3 K9 me 3和HUSH。 与SETDB 1合作促进H3 K9 me 3和异染色质的扩散。 Aim 3（RetroCHMP 3 Blocks to Viral Dissemination）的研究将建立在我们最近发现的retroCHMP 3蛋白的基础上， 其是天然存在的因子，其有效地抑制使用ESCRT途径的包膜病毒的释放， 包括HIV-1。我们的目标是通过增加蛋白质表达来优化小鼠细胞系中的retroCHMP 3效力。 稳定性、表达和限制活性，然后产生表达最佳构建体的转基因小鼠，并测试 对包膜病毒的广泛抗性。 Aim 4（病毒启发设计的输送系统）的研究将产生新的病毒启发技术平台， 细胞间递送是简单、稳健和可控的，其利用了：1）最近的发展， 计算蛋白质设计，2）我们以前在利用HIV-1组装原理开发 纳米粒子，可以指导自己的自我组装和芽从细胞，和3）我们的新进展， 将跨膜蛋白导入这些颗粒中。