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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：
10391770
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/10391770
关键词：
Acyclovir Address 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 CD8-Positive T-Lymphocytes Cell Line Cell physiology Cells Code Cornea Data Dendritic Cells Dendritic cell activation Development Disease Disease Progression Disease model Double Stranded DNA Virus Drug resistance Exhibits Experimental Designs Eye Eye Infections 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 Measures Mediating Messenger RNA MicroRNAs Morbidity - disease rate Mus Myeloid Cells Natural Killer Cells Oligonucleotides Outcome Pathogenesis Pathology Pathway interactions Pattern Persons Production Proteins RNA Regulation Reporting Resolution Role Severities Severity of illness Sorting - Cell Movement Supporting Cell Suspensions Swab T cell response T-Cell Activation T-Lymphocyte Therapeutic Tissues Topical application Transcript Tropism Tumor-infiltrating immune cells United States Untranslated RNA Vaccines Viral Viral Antigens Viral Genes Viral Genome Viral Load result Virion Virus Virus Activation Virus Replication Work analog cytokine cytotoxic defense response design functional disability immune function immunoregulation in vitro Assay in vivo inhibitor insight latent infection lytic gene expression macrophage 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.

1 型单纯疱疹病毒 (HSV-1) 是一种高度传播的感染，在全世界都很常见和流行。相似的与其他人类疱疹病毒 (HHV) 不同，HSV-1 在宿主体内维持终生潜伏期，需要通过以下方式进行免疫逃避：各种复杂的机制。尽管 HSV-1 配备了大量 (>80) 的蛋白质编码基因，但潜伏期病毒基因表达的普遍接受的表现是非编码基因的积累转录本和一组 microRNA (miR)。 HSV-1 编码的病毒 miRNA (v-miR) 被证明可以控制病毒和宿主转录本并调节病毒向性、裂解转换、免疫颠覆等。多项研究虽然已经检查了各种细胞系中的 HSV-1 谱，但这些 v-miR 的关键生物学功能仍然未知。所以，我们建议评估：（1）疾病进展和重新激活期间 v-miR 的系统表达动态，（2）通过扰乱免疫细胞功能在发病机制中发挥综合作用，以及（3）通过 v-miR 进行治疗靶向合成寡核苷酸可减轻 HSV-1 介导的眼部疱疹。使用我们建立的小鼠眼模型疱疹，我们将比较原代小鼠和再激活小鼠角膜组织和血液中的 HSV-1 miRNA 谱。识别正向和负向调节 v-miR 可以对宿主与病毒的相互作用产生新的见解。获得的信息将可用于设计治疗性 v-miR 抑制剂以沉默候选 v-miR 功能。合成效果将评估针对候选 v-miR 的寡核苷酸（v-miR 抑制剂）对小鼠眼部疾病进展的影响模型。 V-miR抑制剂（单独或组合）将被局部递送并释放病毒、病毒转录物/基因组、并将测量疾病严重程度评分。接下来，我们将评估v-miRs如何渲染宿主免疫系统功能失调，这是 HSV-1 持续存在所需的一个不可或缺的特征。使用 v-miR 抑制剂，我们将评估是否免疫在体内可恢复浸润和功能。将全面分析病毒感染的免疫细胞亚群使用流式细胞术和单细胞 RNA 测序使用 v-miR 抑制剂治疗动物。另外，转录与抗原加工/呈递途径相关的基因的表达谱，对于有效的抗病毒反应至关重要，将被量化。这将确定 v-miRs 促进眼部免疫逃避的体内机制。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状细胞的处理/呈递以及 T 细胞的激活。表达 V-miR 的骨髓细胞将评估病毒抗原的摄取和加工以及自体 T 辅助细胞 (CD4+) 和 T 细胞毒性的激活 (CD8+) 细胞。此外，我们将评估 v-miR 对 CD4+ T 细胞极化的影响。生成的数据将为现有的知识差距提供重要信息。总体而言，拟议的转化研究侧重于通过调节 v-miRs 在眼部疾病发病机制中的治疗和机制方面免疫细胞反应。