Cargo, biogenesis and functions of extracellular vesicles released during HSV-1 infection

HSV-1感染期间释放的细胞外囊泡的货物、生物发生和功能

• 批准号: 10439839
• 负责人: Maria Kalamvoki
• 金额: $ 48.57万
• 依托单位: UNIVERSITY OF KANSAS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10439839
• 关键词: Afferent Neurons; Alzheimer' s Disease; Benign; Biogenesis; Capsid Proteins; Cell physiology; Cells; Complex; Cytosol; DNA; Data; Disease; Encephalitis; Epithelial Cells; Gene Expression; Gene Silencing; Glycoproteins; HIV-1; Herpes Labialis; Herpes Simplex Infections; Herpesvirus 1; Human; Immune Evasion; Immune response; Infection; Innate Immune Response; Integration Host Factors; Light; Lytic Phase; Mass Spectrum Analysis; Mediating; Mucous Membrane; Names; Natural Immunity; Neurodegenerative Disorders; Neurons; Pathogenesis; Pathway interactions; Population; Primary Infection; Production; Proteins; Reporting; Role; Sensory Ganglia; Severities; Sorting - Cell Movement; Stimulator of Interferon Genes; Stress; Testing; VP 16; Viral; Viral Genes; Viral Genome; Viral Proteins; Virion; Virus; Virus Replication; cell type; design; env Gene Products; extracellular vesicles; in vivo; insight; intercellular communication; mutant; particle; pathogen; response; sensor; vesicular release

Following productive, lytic infection in mucosal epithelial cells located at the portal of entry in the body, herpes simplex virus-1 (HSV-1) establishes a lifelong, silent infection in sensory neurons. The virus is occasionally reactivated due to weakened immune response or stress causing diseases that range in severity from benign cold sores to encephalitis. HSV-1 contributes to exacerbation of neurodegenerative diseases such as Alzheimer's and facilitates infection by other pathogens such as HIV-1. Extracellular vesicles (EVs) are released by all types of cells, as they constitute a major mechanism of intercellular communication, and they can influence recipient cell functions. During HSV-1 infection two populations of EVs are readily detectable. The first population is produced through a tetraspanins biogenesis pathway. These are the most abundant EVs released from HSV-1 infected cells, which are enriched in CD63 tetraspanin and carry host factors such as the STimulator of INterferon Genes (STING), a sensor of DNA in the cytosol. These CD63+ EVs can activate innate immune responses in uninfected recipient cells and suppress a subsequent HSV-1 infection. The second population of EVs is produced through the endosomal sorting complex required for the transport (ESCRT) pathway, thus we named them ESCRT+ EVs. These EVs carry ESCRT components along with selected viral tegument and envelope protein and they appear to have a proviral role. In addition to these two population of EVs, earlier studies have reported the production of capsidless particles during HSV-1 infection composed of tegument and envelope proteins but lacking viral genome. These particles are lighter than HSV-1 virions and are known as L-particles. L-particles resemble the ESCRT+ EVs in that they carry viral proteins and have a proviral effect but are denser than ESCRT+ EVs and enriched in viral proteins. We hypothesize that distinct populations of EVs are released during HSV- 1 infection with different effects on the infection. To test our hypothesis, we have formulated three Aims. In Aim 1, we propose to compare the cargo of ESCRT+ EVs and CD63+ EVs with L-particles produced during HSV-1 infection using a mass spectrometry approach. The results of this analysis will generate important information about messages communicated by infected cells. In Aim 2, we propose to determine EV biogenesis pathways involved in the production of ESCRT+ EVs, CD63+ EVs and L-particles during HSV-1 infection. For this, we will use a gene silencing approach to compromise selected EV biogenesis pathways or we will use mutant viruses to determine the impact of selected viral genes on EV biogenesis. In Aim 3, we propose to determine the type of responses that CD63+ EVs, ESCRT+ EVs and L-particles trigger in recipient cells and determine the consequences to the infection. These studies will provide important information on the biogenesis, cargo, and functions of EVs released by HSV-1 infected cells, which are essential to determine their impact on HSV-1 pathogenesis.

单纯疱疹病毒1型（HSV-1）在位于体内入口处的粘膜上皮细胞中发生生产性溶解性感染后，在感觉神经元中建立终身沉默感染。由于免疫反应减弱或压力，病毒偶尔会重新激活，导致从良性唇疱疹到脑炎的严重疾病。HSV-1导致神经退行性疾病如阿尔茨海默病的恶化，并促进其他病原体如HIV-1的感染。细胞外囊泡（EV）由所有类型的细胞释放，因为它们构成细胞间通讯的主要机制，并且它们可以影响受体细胞功能。在HSV-1感染期间，两个EV群体容易检测。第一个种群是通过四跨膜蛋白生物合成途径产生的。这些是从HSV-1感染的细胞释放的最丰富的EV，其富含CD 63四跨膜蛋白并携带宿主因子，例如干扰素基因的STimulator（STING），细胞溶质中的DNA传感器。这些CD 63 + EV可以激活未感染的受体细胞中的先天免疫应答，并抑制随后的HSV-1感染。第二群EV通过转运所需的内体分选复合物（ESCRT）途径产生，因此我们将其命名为ESCRT+ EV。这些EV携带ESCRT组分沿着选择的病毒被膜和包膜蛋白，并且它们似乎具有前病毒作用。除了这两种EV群体之外，早期的研究已经报道了在HSV-1感染期间产生由被膜和包膜蛋白组成但缺乏病毒基因组的无衣壳颗粒。这些粒子比HSV-1病毒粒子轻，被称为L粒子。L-颗粒类似于ESCRT+ EV，因为它们携带病毒蛋白并具有前病毒效应，但比ESCRT+ EV更致密并富含病毒蛋白。我们假设不同的EV群体在HSV- 1感染期间释放，对感染具有不同的影响。为了验证我们的假设，我们制定了三个目标。在目标1中，我们建议使用质谱方法比较ESCRT+ EV和CD 63 + EV的货物与HSV-1感染期间产生的L-颗粒。这种分析的结果将产生有关受感染细胞所传达的信息的重要信息。在目标2中，我们提出确定在HSV-1感染期间参与ESCRT+ EV、CD 63 + EV和L-颗粒产生的EV生物发生途径。为此，我们将使用基因沉默方法来折衷选定的EV生物发生途径，或者我们将使用突变病毒来确定选定的病毒基因对EV生物发生的影响。在目标3中，我们提出确定CD 63 + EV、ESCRT+ EV和L-颗粒在受体细胞中触发的应答类型，并确定感染的后果。这些研究将提供关于HSV-1感染细胞释放的EV的生物起源、货物和功能的重要信息，这对于确定它们对HSV-1发病机制的影响至关重要。