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

基本信息

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
项目类别：
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

项目摘要

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.

项目概要本项目的研究旨在推进解决 HIV-1 新领域所需的基础问题生物学和医学，包括制定治疗策略、广泛的抗病毒疗法和递送方法将生物疗法转化为靶细胞。为此，项目 3“病毒的多尺度分析和调节”中的研究动力学，将在广泛的分辨率范围内表征 HIV-1 原病毒沉默和重新激活，从活体动物进行高分辨率结构研究，并将开发旨在保护动物免受包膜侵害的新方法病毒感染和设计新的生物制剂输送系统。目标 1（病毒再激活的原位结构）的研究将在多个尺寸和分辨率范围内进行检查，在动物模型中，HIV/SIV 反弹部位的病毒和相关细胞。具体来说，我们将：1）定位和成像重新激活病毒和相关细胞，2) 定义反弹时受感染细胞和邻近细胞的转录谱网站。目标 2（前病毒沉默和重新激活）的研究寻求从分子和结构上了解 HIV-1 如何作用建立并维持沉默。为此，我们将确定重要的结构、相互作用和机制对于延迟，重点关注 H3K9 特异性甲基化酶 SETDB1 和 HUSH，后者定位于 H3K9me3 和与SETDB1合作促进H3K9me3和异染色质的传播。目标 3（RetroCHMP3 阻止病毒传播）的研究将建立在我们最近发现的 RetroCHMP3 蛋白的基础上，它们是天然存在的因素，可有效抑制使用 ESCRT 途径的包膜病毒的释放正在萌芽，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retroCHMP3效力稳定性、表达和限制活性，然后创建表达最佳构建体的转基因小鼠并测试对包膜病毒具有广泛的抵抗力。目标 4（受病毒启发设计的交付系统）的研究将产生新的受病毒启发的技术平台通过利用以下优势，实现简单、稳健且可控的细胞间传递：1）最近的发展计算蛋白质设计，2) 我们之前在利用 HIV-1 组装原理来开发纳米粒子可以指导它们自己的自组装并从细胞中萌芽，3）我们在整合方面的新进展跨膜蛋白进入这些颗粒。