HSV-1 Encoded MicroRNAs in the Pathogenesis and Treatment of Ocular Herpes

HSV-1 编码的 MicroRNA 在眼疱疹发病机制和治疗中的作用

基本信息

批准号：
10569577
负责人：
Afsar Raza Naqvi
金额：
$ 41万
依托单位：
UNIVERSITY OF ILLINOIS AT CHICAGO
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：
2022-03-01 至 2025-12-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10569577
关键词：
Acyclovir Affect Aftercare Animals Antigen Presentation Pathway Antigen-Presenting Cells Antigens Antiviral Agents Antiviral Response Attenuated Autologous B-Lymphocytes Biological Process Blindness Blood CD4 Positive T Lymphocytes CD8B1 gene Cell Line Cell physiology Cells Code Cornea Data Dendritic Cells Development Disease Disease Progression Disease model Double Stranded DNA Virus Drug resistance Exhibits Experimental Designs Eye Eye Infections Eye diseases Family Flow Cytometry Gene Expression Genes Genome Goals Herpes Labialis Herpesviridae Herpesviridae Infections Herpesvirus 1 Herpetic Keratitis Host Defense Human Immune Immune Evasion Immune response Immune system Impairment Infection Infiltration Inflammation Interferon Activation Interferons Keratitis Kinetics Knowledge Lytic Lytic Virus Macrophage Measures Mediating Messenger RNA MicroRNAs Morbidity - disease rate Mus Myeloid Cells Natural Killer Cells Oligonucleotides Outcome Pathogenesis Pathology Pathway interactions Pattern Persons Production Productivity Proteins RNA Regulation Reporting Resolution Role Severities Severity of illness Sorting Supporting Cell Suspensions Swab T cell response T-Cell Activation T-Lymphocyte Therapeutic Tissues Topical application Transcript Transfection Tropism United States Untranslated RNA Vaccines Viral Viral Antigens Viral Genes Viral Genome Viral Load result Viral Physiology Virion Virus Virus Activation Virus Replication Work analog cytokine cytotoxic defense response design functional disability immune cell infiltrate immune function immunoregulation in vitro Assay in vivo inhibitor insight latent infection lytic gene expression member mouse model neutrophil new therapeutic target novel pathogenic virus prevent response seropositive single-cell RNA sequencing therapeutic target translational study uptake virus host interaction

项目摘要

Herpes Simplex Virus-1 (HSV-1), a highly transmissible infection, is common and endemic throughout the world. Similar to other human herpesviruses (HHV), HSV-1 maintains lifelong latency inside host that requires immune evasion through various sophisticated mechanisms. Although HSV-1 is equipped with large repertoire (>80) of protein coding genes, the commonly accepted manifestations of viral gene expression during latency are the accumulation of a noncoding transcript and a set of microRNAs (miR). HSV-1 encoded viral miRNAs (v-miRs) are demonstrated to control expression of both viral and host transcripts and regulate viral tropism, lytic switching, immune subversion, etc. While multiple studies have examined HSV-1 profiles in various cell lines, key biological functions of these v-miRs remain unknown. Therefore, we propose to evaluate: (1) systematic expression dynamics of v-miRs during disease progression and reactivation, (2) comprehensive role in the pathogenesis by perturbing immune cells functions and (3) therapeutic targeting of v-miR by synthetic oligonucleotides to mitigate HSV-1-mediated ocular herpes. Using our established mouse model of ocular herpes, we will compare HSV-1 miRNA profiles in primary and reactivated mice corneal tissues and blood. Identifying positive and negative regulatory v-miRs can yield novel insights into host-virus interaction. The information gained will be used in designing therapeutic v-miR Inhibitors to silence candidate v-miRs functions. Effect of synthetic oligonucleotides (v-miR Inhibitors) targeting candidate v-miRs will be assessed on ocular disease progression in a mice model. V-miR inhibitors (alone or in combination) will be topically delivered and virus release, viral transcript/genome, and disease severity score will be measured. Next, we will evaluate how v-miRs can render host immune system dysfunctional, an integral feature required for HSV-1 persistence. Using v-miR inhibitors, we will assess whether immune infiltration and functions can be restored in vivo. Immune cell subsets will be comprehensively profiled in virus-infected animals treated with v-miR inhibitor using flow cytometry and single cell RNA sequencing. In addition, transcript expression profiles of genes related to antigen processing/presentation pathway, critical for potent antiviral response, will be quantified. This will identify the in vivo mechanisms through which v-miRs can facilitate immune evasion in ocular tissues. Next, we will dissect underlying mechanisms of v-miR-mediated dysregulation of antigen processing/presentation by macrophages and dendritic cells and activation of T cells. V-miR expressing myeloid cells will be assessed for uptake and processing of viral antigens and activation of autologous T helper (CD4+) and T cytotoxic (CD8+) cells. In addition, we will assess the impact of v-miRs on the polarization of CD4+ T cells. The data generated will provide significant information to existing knowledge gaps. Overall, the proposed translational study focuses on identifying the therapeutic and mechanistic aspect of v-miRs in ocular disease pathogenesis through modulation of immune cell responses.

高度可传播的感染是一种普遍且流行的单纯疱疹病毒-1（HSV-1）。相似的对于其他人类疱疹病毒（HHV），HSV-1维持宿主内部终生潜伏期，需要通过各种复杂的机制。尽管HSV-1配备了大量曲目（> 80）的蛋白质编码基因，但潜伏期期间病毒基因表达的通常接受的表现是非编码的积累转录本和一组microRNA（mir）。 HSV-1编码的病毒miRNA（V-MIRS）被证明可以控制病毒和宿主的成绩单以及调节病毒性疗法，裂解转换，免疫颠覆等，而多次研究已经检查了各种细胞系中的HSV-1谱，这些V-MIR的关键生物学功能仍然未知。所以，我们建议评估：（1）V-MIR在疾病进展和重新激活过程中的系统表达动力学，（2）通过扰动免疫细胞功能和（3）V-MIR的治疗靶向通过发病机理中的全面作用合成寡核苷酸可减轻HSV-1介导的眼疱疹。使用我们已建立的眼部鼠标模型疱疹，我们将比较原发性和重新激活的小鼠角膜组织和血液中的HSV-1 miRNA谱。识别正和负调控V-MIR可以对宿主病毒相互作用产生新的见解。获得的信息将用于设计治疗性V-MIR抑制剂以使候选V-MIRS功能保持沉默。合成的效果靶向候选V-MIR的寡核苷酸（V-MIR抑制剂）将在小鼠的眼部疾病进展中进行评估模型。 V-MIR抑制剂（单独或组合）将被局部传递，病毒释放，病毒转录/基因组，将测量疾病的严重程度评分。接下来，我们将评估V-MIR如何使宿主免疫系统功能失调，HSV-1持久性所需的积分功能。使用V-MIR抑制剂，我们将评估免疫可以在体内恢复渗透和功能。免疫细胞子集将在病毒感染中全面介绍用流式细胞仪和单细胞RNA测序用V-MIR抑制剂处理的动物。另外，成绩单与抗原加工/表现途径有关的基因的表达谱，对有效抗病毒反应至关重要，将被量化。这将确定V-MIR可以促进眼部免疫的体内机制组织。接下来，我们将剖析V-MIR介导的抗原失调的潜在机制巨噬细胞和树突细胞的加工/表现以及T细胞的激活。表达髓样细胞的V-MIR将评估病毒抗原的摄取和加工以及自体T辅助器（CD4+）和T细胞毒性的激活（CD8+）细胞。此外，我们将评估V-MIRS对CD4+ T细胞极化的影响。生成的数据将为现有知识差距提供重要信息。总体而言，拟议的翻译研究重点是通过调制免疫细胞反应。