Role of HIV in KSHV oral transmission

HIV 在 KSHV 口腔传播中的作用

• 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
• 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

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.

项目摘要/摘要 Kaposi Sarcoma（KS）仍然是全世界享有艾滋病毒/艾滋病的人最常见的恶性肿瘤之一。 Kaposi肉瘤疱疹病毒（KSHV），也称为人类疱疹病毒8（HHV-8），是KS的因果剂。这 一旦病毒在口腔中，口服粘膜是KSHV感染的第一个靶标。但是，最初的感染 KSHV的粘膜过程从未被研究过，主要是由于缺乏体外模型来概括 KSHV感染动物模型的体内病毒感染和不存在。我们创建了3- 尺寸（3-d）器官培养物作为一种口服粘膜模仿，类似于分层的口服粘膜 对KSHV感染/传播过程的组织学评估的潜力。我们的研究表明KSHV 3-D口腔粘膜模型中的感染通过细胞外囊泡（EV），尤其是外泌体，尤其是外泌体增强 来自HIV患者。从HIV患者的唾液或HIV感染T的培养基中纯化的外泌体 细胞中包含类似的HIV特异性陈述，包括HIV TAR RNA。 KSHV进入靶细胞及其感染力 在原发性和永生的人口腔上皮细胞中，HIV阳性唾液外泌体增加。更远， HIV-增强了3-D器官培养物中的KSHV感染和向3D器官培养的上prapabasal细胞的传播 阳性外泌体。 HIV阳性外泌体增加的KSHV感染性归因于HIV TAR RNA 宿主细胞的囊泡和表皮生长因子受体（EGFR）。 cetuximab抑制EGFR，A 与EGFR的单克隆抗体，阻断了HIV阳性外泌体增强的KSHV感染。在一起， 这些数据导致我们假设HIV阳性外泌体促进KSHV感染和通过 口腔途径，并负责增加艾滋病毒患者KSHV感染的发病率。测试 该假设，我们计划1）通过3-D评估通过口服途径的KSHV传播和感染 与外周血单核细胞（PBMC）结合的口服粘膜和扁桃体组织的培养物作为 以及上皮 - 内皮模型。 KSHV感染和通过上皮屏障传播到 将评估对HIV+唾液外泌体的免疫和内皮细胞； 2）描述机制 HIV阳性外泌体使用3-D组织模型促进KSHV口腔传播。我们将申请 单细胞RNASEQ鉴定口腔粘膜和扁桃体3-D组织转录组的细胞特异性变化 响应HIV+外泌体的KSHV感染后的模型； 3）阐明依赖EGFR的作用 HIV阳性外泌体增强KSHV传播中有丝分裂原激活的蛋白激酶激活。成功 拟议的研究将提高我们对分子水平的KSHV口腔传播的理解，并 开发潜在的新型治疗策略的基本机制。