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：
9199202
负责人：
KEITH R JEROME
金额：
$ 22万
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 2018-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9199202
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 Herpes Simplex Infections Herpesviridae Infections Herpesvirus Type 3 Homing Human Papillomavirus Immunohistochemistry Individual Induced Mutation Infection Infiltration Inflammatory Inflammatory Response Injection of therapeutic agent Laboratories Latent Virus Latent virus infection phase 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 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 Latency 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）感染仍然是一个常见的严重问题，与原发性感染相关。临床疾病。我们提议使用霍姆底核酸酶（HES）通过靶向病毒基因的靶向诱变来禁用潜在感染神经元的HSV，而病毒性重新激活和复制的来源则是消除我们消除潜在潜伏感染的方法。。 B病毒，艾滋病毒或人乳头瘤病毒在SA1中：评估HSV DNA在Mousel Offection中的疗效，使用了我们的潜在HSV感染的小鼠模型，以使HSV诱变HSV并破坏其反应性：评估和优化hSVO DNA的SAVO在SV i nfection的小鼠模型中，我们将我们的潜在HSV感染来评估和优化HE Exposser的体内耐受性和安全性。开发更大规模动物研究的信息必不可少的，这是对诊所的某种作恋方法。