Defective Viral genomes in RSV pathogenesis

摘要 我们的实验室最近发现了缺陷病毒基因组(DVGs)的关键作用，它是在 大多数病毒感染，作为抗病毒免疫的主要诱导者，对许多呼吸道病毒感染做出反应。 重要的是，检测感染呼吸道合胞病毒的儿童呼吸道分泌物中的DVGs (RSV)与强烈的抗病毒反应的证据相关，支持积累的证据表明 DVGs在调节RSV致病中的作用。然而，对其生物学和临床影响的研究 DVG一直很慢，部分原因是缺乏适当的技术和机制的细节 调节DVG的产生以及与宿主的相互作用仍不清楚。在这里，我们建议利用 重要的技术进步，包括我们从全长病毒基因组中区分DVG的能力 单细胞水平，以显著提高我们对DVG在感染过程中的生成和活动的理解 RSV。这种病原体是婴儿住院的主要原因，可导致严重疾病，甚至 老年人和免疫受抑者的死亡。我们的主要假设是DVG是主要的 人类病毒致病的调节剂及其免疫刺激活性可被利用 尽量减少与病毒相关的疾病。在这项提案中，我们将跟进挑战 目前的范式是DVG在病毒复制期间随机启动，并将定义病毒 调节RSV DVG生成的签名，从而推进操纵DVG的策略 用于研究的感染期间的内容和潜在的治疗收益(目标1)。此外，我们还将利用 我们从细胞内的全长病毒基因组中区分DVG的能力，以了解独特的DVG宿主 调节RSV感染期间抗病毒免疫诱导的相互作用(目标2)。最后，我们将使用 我们随时可以获得独特的临床样本队列，以评估DVGs对临床结果的影响 并建立DVG水平、疾病严重程度和宿主反应之间的关系 人类(目标3)。

Summary Our laboratory recently revealed a critical role for defective viral genomes (DVGs), which are generated during most viral infections, as primary inducers of antiviral immunity in response to many respiratory viral infections. Importantly, detection of DVGs in respiratory secretions from children infected with respiratory syncytial virus (RSV) correlates with evidence of strong antiviral responses, supporting accumulating evidence of a critical role for DVGs in regulating the pathogenesis of RSV. However, study of the biology and clinical impact of DVGs has been slow due, in part, to lack of appropriate technology, and details of the mechanisms that regulate DVG generation and interaction with the host remain unknown. Here we propose to take advantage of important technical advances, including our ability to distinguish DVGs from full-length viral genomes at a single cell level, to significantly improve our understanding of DVG generation and activity during infection with RSV. This pathogen is the major cause of hospitalizations of infants and can cause severe illness and even death in the elderly and the immunosuppressed. Our overarching hypothesis is that DVGs are primary modulators of viral pathogenesis in humans and that their immunostimulatory activity can be harnessed to minimize viral-associated disease. In this proposal, we will follow up on preliminary data challenging the current paradigm that DVG generation initiates randomly during virus replication, and will define viral signatures that regulate the generation of RSV DVGs, thereby advancing strategies to manipulate the DVG content during infection for research and potential therapeutic gain (Aim 1). In addition, we will take advantage of our ability to distinguish DVGs from full-length viral genomes within a cell to understand unique DVG-host interactions that regulate the induction of antiviral immunity during RSV infection (Aim 2). Lastly, we will use unique cohorts of clinical samples readily available to us, to assess the impact of DVGs on the clinical outcome of RSV infection and establish the relationship between DVG levels, disease severity, and host response in humans (Aim 3).