Genetic approaches to discover host factors critical to dengue virus infection

描述（由申请人提供） 摘要：登革热是一种迅速出现的疾病，目前在亚洲、太平洋、美洲、非洲和加勒比地区至少100个国家流行。它每年造成估计1亿例登革热病例，其中症状从轻度发烧到严重的登革出血热约250.000例。登革热是一个主要的全球性健康问题，因为病毒及其蚊子媒介的地理分布不断扩大，流行病的频率增加，多种血清型的共同传播以及在新地区出现严重衰弱的登革出血热。目前还没有批准的疫苗或抗病毒治疗可用于登革热病毒感染。对决定登革热病毒复制和传播的宿主功能的详细了解可能会加速抗病毒药物的开发。我们最近在人类细胞中开发了单倍体遗传筛选，作为进行功能丧失研究的有效方法。使用插入诱变在一个单倍体人类细胞系，我们已经创建了细胞库中的突变，在大多数基因。我们已经证明，这种方法可用于识别不同病原体（包括埃博拉病毒和艰难梭菌）所需的宿主基因。在这份提案中，我们的目标是找到关键的 登革病毒用来建立成功感染的宿主因子。我们将系统地询问参与宿主抗病毒防御的基因的功能，我们将探索这些遗传因素是否可以预测感染个体症状严重程度的显着多样性。我们希望这些研究将揭示登革病毒如何操纵宿主细胞功能以进行复制和传播以及如何逃避先天免疫反应的分子机制。 公共卫生相关性：登革热是一种蚊媒传染病，近几十年来，由于其症状的严重性和病毒的地理传播范围的扩大，登革热已成为一个主要的国际公共卫生问题。深入了解病毒-宿主相互作用的机制将有助于设计特异性抗病毒治疗，并将提高我们对登革病毒发病机制的认识。

DESCRIPTION (Provided by the applicant) Abstract: Dengue is a rapidly emerging disease that is now endemic in at least 100 countries in Asia, the Pacific, the Americas, Africa, and the Caribbean. Annually it causes an estimated 100 million cases of dengue fever, in which symptoms range from a mild fever to severe dengue hemorrhagic fever in approximately 250.000 cases. Dengue is a major global health problem because of the expanding geographical distribution of both the virus and its mosquito vector, the increased frequency of epidemics, the co-circulation of multiple serotypes and the emergence of severely debilitating dengue hemorrhagic fever in new areas. Currently there is no approved vaccine or antiviral treatment available for dengue viral infection. A detailed understanding of the host functions that determine dengue virus replication and spread may accelerate the development of antiviral drugs. We have recently developed haploid genetic screens in human cells as an efficient approach to perform loss-of-function studies. Using insertional mutagenesis in a haploid human cell line, we have created cell libraries with mutations in most genes. We have shown that this approach can be used to identify host genes required for different pathogens including Ebola virus and Clostridium difficile. In this proposal we aim to find critical host factors that dengue virus uses to establish a successful infection. We will systematically interrogate the function of the genes involved in host antiviral defenses, and we will explore whether these genetic factors can predict the remarkable diversity in symptom severity of infected individuals. We expect these studies will uncover molecular mechanisms of how dengue virus manipulates host cell functions for its replication and spread and how it escapes innate immune responses. Public Health Relevance: Dengue is a mosquito-borne infection that in recent decades has become a major international public health concern due to the severity of its symptoms and the expanding geographical spread of the virus. Detailed mechanistic insight in virus-host interactions will enable the design of specific antiviral therapies and will advance our knowledge of dengue virus pathogenesis.