Selectively Replicating Trojan Virus Vectors as Programmable CRISPR-Based Antiviral Therapies

选择性复制特洛伊病毒载体作为基于 CRISPR 的可编程抗病毒疗法

• 批准号: 10196367
• 负责人: Michael Paul Phelps
• 金额: $ 61.2万
• 依托单位: WASHINGTON STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-10 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10196367
• 关键词: 2019-nCoV; Active Sites; Animal Model; Animals; Antiviral Agents; Antiviral Therapy; Area; Biological Models; Biology; COVID-19; COVID-19 pandemic; COVID-19 therapeutics; COVID-19 treatment; CRISPR therapeutics; Cell model; Cells; Clustered Regularly Interspaced Short Palindromic Repeats; Communicable Diseases; Coronavirus; Development; Disease; Disease Outbreaks; Ebola; Economics; Effectiveness; Engineering; Foundations; Future; Genome; Genome Components; Genome engineering; Goals; Hepatitis; Human; Immune; Immunity; Individual; Influenza; Intervention; Invaded; Laboratories; Methods; Mus; Nature; Nucleic Acids; Pathogenicity; Patients; Population; Pre-Clinical Model; Protein Engineering; Proteins; RNA Interference; Regulation; Research; Resistance; Resources; Ribonucleases; SARS coronavirus; SARS-CoV-2 antiviral; SARS-CoV-2 infection; Series; Societies; Structure; System; Technology; Testing; Therapeutic; Time; Tissues; Vaccine Therapy; Vaccines; Viral; Viral Genome; Viral Proteins; Viral Vector; Virus; Virus Diseases; Virus Replication; acquired immunity; base; combat; coronavirus therapeutics; critical period; design; economic impact; effective therapy; fighting; gene therapy; genome editing; humanized mouse; in vivo; in vivo evaluation; insight; interest; mouse model; novel; novel coronavirus; novel strategies; particle; pathogen; pathogenic virus; prevent; programs; protein complex; protein structure; research clinical testing; small molecule; social; success; technology development; therapy development; tool; transmission process; vector; viral transmission

PROJECT SUMMARY The COVID-19 pandemic has highlighted our long-standing vulnerability to new viral infectious diseases with which there is no acquired immunity. While vaccines and antiviral therapies can eventually be developed to treat many viral infectious diseases, these interventions require significant time and resources to acquire. This results in a critical period of time where there are no therapeutic options to slow the spread of the virus, besides physical countermeasures that have dramatic economic and social consequences. The truly alarming insight is that all of the vaccines and therapies we develop now against SARS-CoV-2 will be useless against the next cycle of viral outbreaks (e.g., influenza, ebola, etc.). There is a paramount need to fundamentally transform our approach to combating emerging viral diseases by developing antiviral strategies that can be rapidly deployed at the onset of a new viral outbreak. We propose a revolutionary new viral-antiviral technology that has the potential to target emerging viral pathogens at all stages of the disease outbreak cycle. This includes targeting viral pathogens in animal hosts prior to human transmission, preventing viral infections in healthy individuals, and treating ongoing viral infections. This new “Trojan virus” technology uses engineered viral vectors that imitate viral pathogens, yet contain potent CRISPR antiviral machinery that degrades pathogenic viral particles. These Trojan virus vectors have an incomplete viral genome that can selectively replicate only in previously infected cells by hijacking viral derived proteins, which it uses to multiply and spread throughout the infected areas of the body. The spread of the Trojan virus acts to prevent viral infection in healthy tissue by targeting invading viral particles, while at the same time suppressing active sites of viral infection. The integration of CRISPR antiviral technology into Trojan virus vectors allows the system to be reprogrammed to target new viral strains without extensive protein engineering or clinical testing, facilitating the rapid mobilization of the technology during viral disease outbreaks. The proposed research will focus on developing SARS-CoV-2 Trojan virus technology as a therapeutic option for active viral infections. The research will use engineered non-infectious cellular model systems to evaluate SARS-CoV-2 Trojan virus genome designs that can selectively replicate only in previously infected cells. We will determine the optimal strategy for incorporating CRISPR antiviral technology into the SARS-CoV-2 Trojan virus vectors, while identifying key CRISPR vulnerabilities in the SARS-CoV-2 virus. To validate that the genome engineering principles developed for the SARS-CoV-2 Trojan virus therapy can mitigate active viral infections, we will duplicate the approach to target the mouse hepatitis coronavirus (MHV-A59), tracking the effectiveness of the Trojan virus in murine model systems. If successful, our research will generate a functional SARS-CoV-2 Trojan virus therapy, validating the genome design in preclinical model systems, to establish the foundation for commercial development of the technology for use in the current and future SARS- CoV outbreaks.

项目摘要 COVID-19大流行强调了我们长期以来对新病毒感染疾病的脆弱性 没有获得的免疫力。虽然有时可以开发疫苗和抗病毒疗法来治疗 这些病毒感染疾病，这些干预措施需要大量的时间和资源才能获得。这结果 在关键时期，除了物理 具有巨大经济和社会后果的对策。真正令人震惊的见解是 我们现在针对SARS-COV-2开发的疫苗和疗法将无用 病毒爆发（例如，影响力，埃博拉病毒等）。从根本上改变我们的方法有至高无上的需求 通过制定可以在开始时快速部署的抗病毒药策略来打击新兴病毒疾病 新病毒爆发。我们提出了一种革命性的新病毒抗病毒技术，有可能针对 在疾病暴发周期的所有阶段，新兴的病毒病原体。这包括针对病毒病原体 动物宿主在人类传播之前，预防健康个体的病毒感染，并治疗正在进行的 病毒感染。这项新的“特洛伊木马病毒”技术使用了模仿病毒病原体的工程病毒载体， 然而，包含潜在的CRIS抗病毒机制，可降解致病病毒颗粒。这些木马病毒 向量具有不完全的病毒基因组，可以选择性地在先前感染的细胞中选择性地复制 劫持病毒衍生的蛋白质，它用于繁殖并散布在身体的整个感染区域。 木马病毒的扩散可通过靶向入侵病毒颗粒来防止健康组织中的病毒感染， 同时抑制病毒感染的活跃部位。 CRISPR抗病毒技术的整合 进入特洛伊木马病毒载体允许对系统进行重新编程以靶向新的病毒株而无需广泛 蛋白质工程或临床测试，支持病毒疾病期间技术的快速动员 爆发。拟议的研究将着重于开发SARS-COV-2 Trojan病毒技术作为一种 主动病毒感染的治疗选择。该研究将使用工程的非感染性蜂窝模型 评估SARS-COV-2 Trojan病毒基因组设计的系统，这些设计只能在先前选择性地复制 感染细胞。我们将确定将CRISPR抗病毒技术纳入纳入的最佳策略 SARS-COV-2 Trojan病毒载体，同时识别SARS-COV-2病毒中的关键CRISPR脆弱性。到 验证为SARS-COV-2 Trojan病毒疗法开发的基因组工程原理可以减轻 主动病毒感染，我们将复制针对小鼠肝炎冠状病毒（MHV-A59）的方法， 跟踪特洛伊木就在鼠模型系统中的有效性。如果成功，我们的研究将产生 一种功能性SARS-COV-2 Trojan病毒疗法，验证临床前模型系统中的基因组设计 建立该技术的商业开发基础，用于当前和未来的SARS- COV爆发。