Role of HIV in KSHV oral transmission

HIV 在 KSHV 口腔传播中的作用

基本信息

批准号：
10491098
负责人：
Ge Jin
金额：
$ 61.04万
依托单位：
CASE WESTERN RESERVE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-09-20 至 2026-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10491098
关键词：
3-Dimensional AIDS/HIV problem Aging Animal Model Body Fluids Cells Cetuximab Culture Media Data Development Endothelial Cells Endothelium Epidermal Growth Factor Receptor Epithelial Epithelial Cells Extracellular Space Gene Expression Goals HIV HIV Infections HIV Seronegativity HIV Seropositivity Herpesviridae Infections Histologic Human Human Herpesvirus 8 Immune Incidence Infection Inflammatory Kaposi Sarcoma Lead Link Malignant Neoplasms Mediating Methods Mitogen-Activated Protein Kinases Modeling Molecular Monoclonal Antibodies Morbidity - disease rate Mucous Membrane Multicentric Angiofollicular Lymphoid Hyperplasia Oncogenic Oral Oral cavity Oral mucous membrane structure Outcome Pathway interactions Patients Peripheral Blood Mononuclear Cell Persons Play Population Predisposition Process Publishing RNA Receptor Signaling Research Response Elements Role Route Saliva Stratum Basale Syndrome System T-Lymphocyte TLR3 gene Testing Tissue Model Tissues Tonsil Transactivation Vascular Endothelium Vesicle Viral Viral Genes Virus Virus Diseases cell type cytokine exosome extracellular vesicles gammaherpesvirus in vitro Model in vivo infection risk lytic replication mortality nanoparticle novel therapeutic intervention oral cavity epithelium oral infection prevent primary effusion lymphoma response single-cell RNA sequencing success transcriptome transmission process

项目摘要

Project Summary/Abstract Kaposi sarcoma (KS) remains one of the most common malignancies in people living with HIV/AIDS worldwide. Kaposi sarcoma herpesvirus (KSHV), also called human herpesvirus-8 (HHV-8), is the causal agent for KS. The oral mucosa is the first target of KSHV infection once the virus is in the oral cavity. However, the initial infection process of the mucosa by KSHV has never been studied, mainly due to lack of an in vitro model to recapitulate the viral infection in vivo and nonexistence of an animal model for KSHV infection. We have created the 3- dimentional (3-D) organotypic culture as an oral mucosal mimic resembling a stratified oral mucosa with the potential for histological assessment of the KSHV infection/transmission process. Our studies show that KSHV infection in the 3-D oral mucosa model is enhanced by saliva extracellular vesicles (EVs), particularly exosomes, from HIV patients. Exosomes purified from the saliva of HIV patients or from culture media of HIV-infected T cells contain similar HIV-specific cargos, including HIV TAR RNA. KSHV entry into target cells and its infectivity are increased by HIV-positive saliva exosomes in primary and immortalized human oral epithelial cells. Further, KSHV infection and transmission to the suprabasal cells in the 3-D organotypic culture are enhanced by HIV- positive exosomes. Increased KSHV infectivity by HIV-positive exosomes is attributed to the HIV TAR RNA within the vesicles and epidermal growth factor receptor (EGFR) of host cells. Inhibition of EGFR by Cetuximab, a monoclonal antibody to EGFR, blocks KSHV infection enhanced by HIV-positive exosomes. Taken together, these data lead us to hypothesize that HIV-positive exosomes promote KSHV infection and transmission through the oral route and are responsible for increased incidence of KSHV infection in people living with HIV. To test this hypothesis, we plan to 1) assess KSHV transmission and infection through the oral route with the 3-D cultures of oral mucosal and tonsil tissues incorporated with peripheral blood mononuclear cells (PBMCs) as well as the epithelium-endothelium model. KSHV infection and transmission through the epithelial barrier to immune and endothelial cells in response to HIV+ saliva exosomes will be assessed; 2) delineate the mechanism by which HIV-positive exosomes promote KSHV oral transmission using the 3-D tissue models. We will apply single cell RNAseq to identify cell-specific changes in transcriptome of the oral mucosal and tonsil 3-D tissue models following KSHV infection in response to HIV+ exosomes; and 3) elucidate the role of EGFR-dependent mitogen-activated protein kinase activation in enhanced KSHV transmission by HIV-positive exosomes. Success of the proposed research will advance our understanding of KSHV oral transmission at the molecular level and the underlying mechanisms for developing potential novel therapeutic strategies.

项目概要/摘要卡波西肉瘤（KS）仍然是全世界艾滋病毒/艾滋病患者最常见的恶性肿瘤之一。卡波西肉瘤疱疹病毒 (KSHV)，也称为人类疱疹病毒 8 (HHV-8)，是 KS 的病原体。这一旦病毒进入口腔，口腔粘膜就是KSHV感染的第一个目标。然而，最初的感染 KSHV 对粘膜的作用从未被研究过，主要是由于缺乏体外模型来概括体内病毒感染且不存在 KSHV 感染动物模型。我们创建了 3- 三维（3-D）器官型培养物作为口腔粘膜模拟物，类似于分层口腔粘膜，对 KSHV 感染/传播过程进行组织学评估的潜力。我们的研究表明 KSHV 唾液细胞外囊泡（EV），特别是外泌体，增强了 3D 口腔粘膜模型中的感染，来自艾滋病毒患者。从 HIV 患者唾液或 HIV 感染者的培养基中纯化的外泌体细胞含有类似的 HIV 特异性货物，包括 HIV TAR RNA。 KSHV进入靶细胞及其感染性原代和永生化人类口腔上皮细胞中的 HIV 阳性唾液外泌体增加了这种水平。更远， HIV 增强了 KSHV 感染并传播至 3D 器官型培养物中的基底上细胞阳性外泌体。 HIV 阳性外泌体增加 KSHV 感染性归因于体内的 HIV TAR RNA 宿主细胞的囊泡和表皮生长因子受体（EGFR）。西妥昔单抗 (Cetuximab) 对 EGFR 的抑制针对 EGFR 的单克隆抗体，可阻断 HIV 阳性外泌体增强的 KSHV 感染。综合起来，这些数据使我们推测 HIV 阳性外泌体通过以下方式促进 KSHV 感染和传播：口服途径，是 HIV 感染者 KSHV 感染发病率增加的原因。测试根据这一假设，我们计划 1) 使用 3-D 评估 KSHV 通过口腔途径的传播和感染口腔粘膜和扁桃体组织与外周血单核细胞（PBMC）的培养物以及上皮-内皮模型。 KSHV 感染并通过上皮屏障传播将评估免疫细胞和内皮细胞对 HIV+ 唾液外泌体的反应； 2）描绘机制 HIV 阳性外泌体利用 3D 组织模型促进 KSHV 口腔传播。我们将申请单细胞 RNAseq 用于识别口腔粘膜和扁桃体 3-D 组织转录组的细胞特异性变化 KSHV 感染后响应 HIV+ 外泌体的模型； 3) 阐明 EGFR 依赖性的作用 HIV 阳性外泌体增强 KSHV 传播过程中丝裂原激活蛋白激酶的激活。成功拟议的研究将增进我们对 KSHV 口腔传播在分子水平上的理解开发潜在的新型治疗策略的潜在机制。