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In vivo inactivation of latent HSV by endonuclease-mediated mutagenesis

通过核酸内切酶介导的诱变体内潜伏 HSV 灭活

基本信息

批准号：
9035463
负责人：
KEITH R JEROME
金额：
$ 26.4万
依托单位：
FRED HUTCHINSON CANCER RESEARCH CENTER
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-01-01 至 2017-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9035463
关键词：
Acute Adverse effects Afferent Neurons Animals Antiviral Therapy Biology Cell Death Cells Chronic Cleaved cell Clinic Clinical DNA DNA Sequence Data Dependovirus Development Disease Doxycycline Enzymes Genes Genome Goals HIV Hepatitis B Virus Herpesviridae Infections Herpesvirus 1 Herpesvirus Type 3 Homing Human Papillomavirus Immunohistochemistry Individual Induced Mutation Infection Infiltration Inflammatory Inflammatory Response Injection of therapeutic agent Laboratories Latent Virus Life Measures Mediating Methods Modeling Modification Morbidity - disease rate Mus Mutagenesis Mutate Mutation Neurons Pathway interactions Phase Positioning Attribute Primary Infection Production Recurrence Recurrent disease Safety Sampling Sequence Analysis Simplexvirus Site Site-Directed Mutagenesis Source Structure of trigeminal ganglion Technology Testing Tetracyclines Therapeutic Time Toxic effect Vaccines Vibrissae Viral Viral Genes Viral Genome Viral load measurement Viral reservoir Virus Virus Diseases Virus Replication Work base clinically relevant curative treatments digital endonuclease experience in vivo innovation knowledge base latent infection mouse genome mouse model next generation sequencing novel therapeutic intervention pre-clinical promoter public health relevance reactivation from latency research clinical testing spinal nerve posterior root tool transgene expression transmission process vector viral DNA virology virus pathogenesis

项目摘要

DESCRIPTION (provided by applicant): Herpes simplex virus (HSV) infections remain a common, serious problem associated with significant morbidity. After primary infection HSV establishes latency, which is not eliminated by current antiviral therapy, and latent virus is the source for viral reactivation and the recurrence of clinical disease. Despite much effort, a vaccine remains elusive. Therefore, there is a need for a new therapeutic approach that would cure latent HSV infection. Our hypothesis is that the emergence of designer rare-cutting endonucleases as powerful tools for directed genome modification offers the unique ability to selectively target, cleave, and disrupt essential viral DNA sequences within living cells. Here, we propose to use homing endonucleases (HEs) to disable HSV in latently infected neurons by targeted mutagenesis of essential viral genes, eliminating the source of viral reactivation and replication. The goal of this project is to optimize and evaluate our approach to eliminate latent HSV infection in vivo using a murine model of HSV latent infection. Moreover, our results will be directly applicable in efforts to cure varicella zoster virus, another alphaherpesvirus that like HSV establishes latency in sensory neurons. Furthermore, the data generated will be highly relevant to the development of a cure of other chronic or latent viral infections such as hepatitis B virus, HIV, or human papillomavirus. In SA1: Evaluate the efficacy in vivo of HE mutagenesis of HSV DNA in a mouse model of latent HSV infection, we will use our mouse model of latent HSV infection to test the ability of HEs to mutagenize HSV and disrupt its ability to reactivate. I SA2: Evaluate and optimize the safety in vivo of HE mutagenesis of HSV DNA in a mouse model of latent HSV infection, we will use our mouse model of latent HSV infection to evaluate and optimize the in vivo tolerability and safety of HE exposure. This project is expected to demonstrate the feasibility of our therapeutic approach directed towards the elimination of HSV pathogenesis in vivo, and to provide critical information for the development of a larger scale animal study necessary to bring this new therapeutic approach to the clinic.

描述（由申请人提供）：单纯疱疹病毒（HSV）感染仍然是一种常见的严重问题，与显著的发病率相关。单纯疱疹病毒初次感染后会出现潜伏期，目前的抗病毒治疗无法消除，潜伏期病毒是病毒再激活和临床疾病复发的根源。尽管做了很多努力，但疫苗仍然难以捉摸。因此，需要一种新的治疗方法来治愈潜伏的HSV感染。我们的假设是，作为定向基因组修饰的强大工具的设计师稀有切割核酸内切酶的出现提供了选择性靶向、切割和破坏活细胞内的必需病毒DNA序列的独特能力。这里我们提出使用归巢核酸内切酶（HE）通过必需病毒基因的靶向诱变来使潜伏感染的神经元中的HSV失活，从而消除病毒再活化和复制的来源。本项目的目标是优化和评估我们的方法，以消除潜伏的HSV感染在体内使用HSV潜伏感染的小鼠模型。此外，我们的研究结果将直接适用于治疗水痘带状疱疹病毒，另一种α疱疹病毒，如HSV在感觉神经元中建立潜伏期。此外，生成的数据将与肝炎等其他慢性或潜伏病毒感染的治疗方法的开发高度相关 B病毒、HIV或人乳头瘤病毒。在SA 1中：在潜伏性HSV感染的小鼠模型中评价HSV DNA的HE诱变的体内效力，我们将使用我们的潜伏性HSV感染的小鼠模型来测试HE诱变HSV并破坏其再活化能力的能力。I SA2：在潜伏性HSV感染的小鼠模型中评价和优化HSV DNA的HE诱变的体内安全性，我们将使用我们的潜伏性HSV感染的小鼠模型来评价和优化HE暴露的体内耐受性和安全性。该项目预计将证明我们的治疗方法的可行性，旨在消除体内HSV发病机制，并为开发更大规模的动物研究提供关键信息，这些研究是将这种新的治疗方法推向临床所必需的。