Antagonizing miRNAs in a strategy to cure HSV latency

拮抗 miRNA 来治愈 HSV 潜伏期

基本信息

批准号：
8510128
负责人：
DONALD M COEN
金额：
$ 26.51万
依托单位：
HARVARD MEDICAL SCHOOL
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-03-01 至 2015-02-28
项目状态：
已结题

项目摘要

DESCRIPTION (provided by applicant): The long-term objective of this project is to develop a novel intervention strategy to clear latent infections by herpes simplex virus (HSV). Latency is the most fascinating aspect of HSV biology and the most vexing aspect of HSV disease -- latency is why HSV infections have yet to be cured. The intervention strategy proposed here is: 1) Disrupt latency by antagonizing viral microRNAs (miRNAs) that we hypothesize repress "lytic" viral gene expression in latently infected neurons, using oligonucleotides that are complementary to these miRNAs (anti-miRs); 2) If necessary, further disrupt latency by antagonizing host functions that help maintain latency; and 3) Irreversibly inactivate the resultin replicating HSV using the anti-HSV drug acyclovir. The R21 phase of this project is a collaborative effort of three labs. The first aim is to determine whether selected anti-miRs or selected viral mutations affecting miRNAs or their target sites (Coen lab) derepress lytic gene expression in mouse and rat neuron in vitro models of HSV latency (Leib and Wilson labs). Gene expression will be measured using PCR-based methods (Coen and Wilson labs). Selected mutations will also be tested for their effects on lytic gene expression in an in vivo mouse latency model (Coen lab). The second aim is to test whether these anti-miRs, alone or in at least additive combination with small molecule antagonists of host functions, induce reactivation in the mouse and rat in vitro models, and whether acyclovir can cure latency in such models. The R33 phase of this project (Coen lab) focuses on efforts to cure latency in vivo in a mouse model. The third aim investigates anti-miRs from Aim 2, and comparable HSV mutations for effects on derepression of gene expression in this model. The fourth aim is to test whether these anti-miRs and comparable mutations, alone or in at least additive combination with small molecules from Aim 2 induce reactivation in the mouse model. The fifth aim is to test whether combinations identified in Aim 4 together with acyclovir can cure latency in vivo in the mouse model. These studies will be accompanied by pharmacokinetic assays of anti-miR and compound concentrations in plasma and tissues, and assays of toxicities. The goal is to achieve cure of HSV with minimal toxicity.

描述（由申请人提供）：该项目的长期目标是制定一种新颖的干预策略，以清除单纯疱疹病毒（HSV）的潜在感染。潜伏期是HSV生物学最迷人的方面，也是HSV疾病最令人沮丧的方面 - 潜伏期是HSV感染尚未治愈的原因。此处提出的干预策略是：1）通过对抑制与这些miRNA（抗MIRS）互补的寡核苷酸（抗MIRNA）相辅相成的抑制病毒microRNA（miRNA）来破坏潜伏期的潜伏期； 2）如有必要，通过拮抗有助于保持潜伏期的宿主功能来进一步破坏潜伏期； 3）不可逆转地使结果使用抗HSV药物acyclovir复制HSV。该项目的R21阶段是三个实验室的协作工作。第一个目的是确定在HSV潜伏期的体外模型（Leib和Wilson Labs）的小鼠和大鼠神经元中的选择影响miRNA或其目标位点（Coen Lab）的选定病毒突变（Coen Lab）。基因表达将使用基于PCR的方法（Coen和Wilson Labs）测量。所选突变还将测试其对体内小鼠潜伏期模型（Coen Lab）中裂解基因表达的影响。第二个目的是测试这些抗MIR是否单独或至少与宿主功能的小分子拮抗剂，诱导小鼠和大鼠体外模型中的重新激活以及Acyclovir是否可以在此类模型中治愈潜伏期。该项目的R33阶段（COEN LAB）着重于在小鼠模型中治愈体内潜伏期的努力。第三个目标研究了来自AIM 2的抗MIR，以及在该模型中对基因表达不抑制的影响的可比HSV突变。第四个目的是测试这些抗MIR和可比较的突变是单独的还是至少与AIM 2中的小分子相结合，在小鼠模型中诱导重新激活。第五目的是测试AIM 4与Acyclovir中鉴定的组合是否可以在小鼠模型中治愈体内潜伏期。这些研究将伴随着血浆和组织中抗MIR和复合浓度的药代动力学测定，以及毒性测定。目的是通过最小的毒性来治愈HSV。