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 卡波西肉瘤(KS)是世界范围内HIV/AIDS患者最常见的恶性肿瘤之一。 卡波西肉瘤疱疹病毒(KSHV)又称人类疱疹病毒8型(HHV-8)，是引起KS的病原体。这个 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对粘膜的作用过程从未被研究过，这主要是由于缺乏一个可以概括的体外模型。 体内病毒感染和不存在KSHV感染的动物模型。我们已经创造了3个- 三维(3-D)器官型培养，作为口腔粘膜的模拟物，类似于分层的口腔粘膜，具有 组织学评估KSHV感染/传播过程的可能性。我们的研究表明，KSHV 唾液细胞外小泡(EVS)，特别是外切体，会增强3D口腔粘膜模型中的感染。 从艾滋病患者那里。从HIV患者唾液或HIV感染T细胞培养上清液中纯化的外切体 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 在原代和永生化的人类口腔上皮细胞中，HIV阳性的唾液外切体增加了。此外， 在三维器官培养中，KSHV的感染和向超基底细胞的传播被HIV-1增强。 阳性外显子。HIV阳性外体增加KSHV的感染性归因于其中的HIV TAR RNA 宿主细胞的囊泡和表皮生长因子受体(EGFR)。西妥昔单抗对EGFR的抑制作用 抗表皮生长因子受体的单抗可阻断HIV阳性外切体增强的KSHV感染。加在一起， 这些数据使我们假设HIV阳性的外切体通过 KSHV感染是通过口服途径传播的，是艾滋病毒携带者感染KSHV增加的原因。为了测试 在这一假设下，我们计划1)评估KSHV通过口腔途径的3-D传播和感染 口腔粘膜和扁桃体组织与外周血单个核细胞(PBMC)复合培养 以及上皮-内皮模型。KSHV感染并通过上皮屏障传播到 将评估免疫和内皮细胞对HIV+唾液外切体的反应；2)描述其机制 HIV阳性的外切体通过3D组织模型促进KSHV的口服传播。我们会申请 单细胞RNAseq用于鉴定口腔粘膜和扁桃体三维组织转录组的细胞特异性变化 KSHV感染后对HIV+外切体反应的模型；3)阐明EGFR依赖的作用 丝裂原活化蛋白激酶激活在HIV阳性外切体促进KSHV传播中的作用。成功 这项拟议的研究将在分子水平上促进我们对KSHV口头传播的理解，并 开发潜在的新治疗策略的潜在机制。