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.

项目摘要 在该项目中进行研究的目的是为了解决HIV-1中的新领域所需的基础 生物学和医学，包括制定治疗策略，广泛的抗病毒疗法和交付方法 靶细胞中的生物疗法。到这些目的，项目3中的研究，多尺度分析和病毒的调节 动力学将表征HIV-1病毒的沉默和重新激活在各种分辨率范围内，从生活中 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包膜的新方法 病毒感染和设计新的生物制剂输送系统。 在AIM 1中的研究（病毒重新激活的原位结构）将跨多个大小和分辨率量表检查， 动物模型中HIV/SIV部位的病毒和相关细胞。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）定义反弹处感染和相邻细胞的转录曲线 站点。 AIM 2（前病毒沉默和重新激活）中的研究寻求分子和结构性理解HIV-1 建立和维护沉默。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特异性甲基酶SETDB1，以及Hush，它本地化在H3K9me3和 与SETDB1合作，以促进H3K9me3和异染色质的传播。 在AIM 3中的研究（恢复传播的retrochmp3块）将建立在我们最近发现的retrochmp3蛋白的基础上 这是自然存在的因素，可有效抑制使用ESCRT途径的包膜病毒的释放 萌芽，包括HIV-1。我们的目标是通过增加蛋白质来优化小鼠细胞系中的恢复效力 稳定性，表达和限制活动，然后创建转基因小鼠，表达最佳构建体并测试 对包裹病毒的广泛抗药性。 AIM 4的研究（受病毒启发设计的递送系统）将为病毒启发的新技术平台生成新的技术平台 通过利用以下优势，简单，健壮且可控制的细胞间交付：1）最近的发展 计算蛋白设计，2）我们以前在利用HIV-1组装原理的成功方面取得了成功 可以指导自己的自组装和芽的纳米颗粒，以及3）我们合并的新进步 跨膜蛋白进入这些颗粒。