Rapid Induction of apoptosis against viral infection

描述（由申请人提供）：节肢动物传播的病原体每年导致数百万人死亡。了解媒介对病原体易感性的遗传基础对于制定新的疾病控制策略至关重要。诱导细胞凋亡是一种基本的先天免疫反应的假设已经得到病毒学研究的支持，这些研究表明许多病毒基因的抗凋亡活性对其感染性和/或毒性至关重要。然而，介导病毒感染后诱导细胞凋亡的细胞机制仍然是谜。此外，对培养昆虫细胞的研究表明，要么缺乏凋亡，要么促凋亡反应发生相对较晚，这让人怀疑凋亡作为一种先天免疫的功能意义。利用模拟原生病毒感染途径的蚊子体内模型，我们发现在暴露于DNA/RNA病毒后的几个小时内，促凋亡基因（RIPAG）快速诱导。更重要的是，利用果蝇的遗传工具，我们发现RIPAG和随后的细胞凋亡，负责拒绝病毒基因的表达和阻断/限制感染。RIAPG受损的动物比野生型更容易受到病毒感染。在本提案中，我们试图利用果蝇遗传学和比较基因组学的结合来揭示控制RIPAG的转录机制和调控途径。此外，利用机制分析获得的信息，我们将验证通过增强RIPAG对病毒感染的反应来增强抗病毒感染的先天免疫的假设。最后，我们评估了我们在果蝇中创建的抗病毒构建体的适应性，然后将最强大和最适合的构建体转化为两种蚊子载体，并对转基因抗登革热的有效性进行了初步评估。

DESCRIPTION (provided by applicant): Arthropod-borne pathogens account for millions of death each year. Understanding the genetic basis of vector susceptibility to pathogens is pivotal to novel disease control strategies. The hypothesis that induction of apoptosis is a fundamental innate immune response has been supported by virology studies, which demonstrated that the anti- apoptotic activities of many viral genes are essential for their infectivity and/or virulence However, the cellular mechanism mediating the induction of apoptosis following virus infection remained enigmatic. In addition, studies with cultured insect cells showed that either there is a lack of apoptosis, or the pro-apoptotic response happens relatively late, casting doubt on the functional significance of apoptosis as an innate immunity. Using in vivo mosquito models mimicking native routes of viral infection, we found that there is a rapid induction of pro-apoptotic genes (RIPAG) within a few hours following exposure to DNA/RNA viruses. More importantly, using genetic tools in Drosophila, we showed that the RIPAG, and the ensuing apoptosis, is responsible for denying the expression of viral genes and blocking/limiting the infection. Animals with compromised RIAPG are much more susceptible to viral infection than wild type. In this proposal, we seek to unravel the transcriptional mechanisms and the regulatory pathway(s) controlling RIPAG using a combination of Drosophila genetics and comparative genomics. In addition, utilizing the information obtained through the mechanistic analysis, we will test the hypothesis that increased innate immunity against viral infection may be achieved by enhancing the RIPAG response to viral infection. Finally, we assess the fitness of the antiviral constructs we create in Drosophila, then translate the most powerful and most fit constructs to two mosquito vectors and perform preliminary evaluations of transgene effectiveness against Dengue.