Genetic approaches to discover host factors critical to dengue virus infection

描述(由申请人提供) 摘要：登革热是一种迅速流行的疾病，目前在亚洲、太平洋、美洲、非洲和加勒比海地区至少有100个国家流行。每年它造成的登革热病例估计有1亿例，症状从轻度发烧到严重登革热出血热约有250.000例。登革热是一个主要的全球健康问题，因为病毒及其蚊媒的地理分布不断扩大，流行频率增加，多种血清型共同循环，以及在新地区出现严重削弱人身体健康的登革热出血热。目前还没有获得批准的登革热病毒感染疫苗或抗病毒治疗。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基因筛查，作为进行功能丧失研究的有效方法。在单倍体人类细胞系中使用插入突变，我们已经创建了具有大多数基因突变的细胞库。我们已经证明，这种方法可以用来识别不同病原体所需的宿主基因，包括埃博拉病毒和艰难梭菌。在本提案中，我们的目标是找到关键 登革热病毒用来确定成功感染的宿主因素。我们将系统地询问参与宿主抗病毒防御的基因的功能，并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著差异。我们预计这些研究将揭示登革热病毒如何操纵宿主细胞功能进行复制和传播，以及它如何逃避先天免疫反应的分子机制。 公共卫生相关性：登革热是一种由蚊子传播的传染病，由于其症状的严重性和病毒在地理上的传播范围扩大，近几十年来已成为一个主要的国际公共卫生问题。对病毒-宿主相互作用的详细机制洞察将使特定抗病毒疗法的设计成为可能，并将促进我们对登革病毒发病机制的了解。

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.