Analysis of molecular factors influencing variability of mosquito gut microbiota

 描述(申请人提供)：由蚊子传播的人类疾病是一个重大的公共卫生问题，迫切需要新的和创新的方法来防止媒介传播的疾病的传播。由于这些病原体必须感染蚊子才能传播给人类，因此有可能通过预防来中断传播周期 蚊子的感染。最近的研究表明，蚊子肠道中的微生物可以影响导致疟疾和登革热等疾病的病原体的传染性。此外，某些种类的细菌已被证明对这些和其他媒介传播的疾病具有直接的抗病原体活性。之前的研究表明，蚊子肠道微生物区系可能是高度可变的。本提案旨在确定关键的遗传因素，并预测埃及伊蚊肠道微生物区系大小和组成的差异背后的机制。这将通过比较不同肠道微生物区系大小和组成的伊蚊菌株的基因表达模式来实现。通过使用全基因组基因表达微阵列分析，将以无偏见的方式识别感兴趣的基因，并使用基因特异性基因表达研究和RNAi敲除功能分析确认它们在塑造肠道微生物区系中的作用。还将测试候选基因对波多黎各登革热流行地区采集的蚊子肠道微生物区系的影响。这项工作将提供对塑造蚊子肠道微生物区系的分子因素和机制的重要洞察，并有助于其变异性，这已被证明影响媒介能力。这些信息将促进我们对蚊子肠道免疫的了解，并将对开发基于蚊子肠道微生物区系的新型疾病控制策略具有重要意义。

 DESCRIPTION (provided by applicant): Human diseases vectored by mosquitoes are a significant public health concern, and novel and innovative methods to prevent the spread of vector borne diseases are in high demand. Because these pathogens must infect a mosquito in order to be transmitted to humans, it is possible to interrupt the transmission cycle by preventing infections in mosquitoes. Recent research has demonstrated that the microbes in the mosquito gut can influence the infectivity of pathogens causing diseases such as malaria and dengue. Moreover, certain species of bacteria have been shown to have direct anti-pathogen activity on these and other vector borne diseases. Previous work has revealed that the mosquito gut microbiota can be highly variable. The present proposal aims to determine key genetic factors and predict mechanisms that underlie variability in the size and composition of the gut microbiota of Aedes aegypti mosquitoes. This will be accomplished by comparing the gene expression patterns across strains of Aedes that vary in gut microbiota size and composition. Genes of interest will be identified in an unbiased fashion, through the use of whole-genome gene expression microarray analysis, and confirmed for their role in shaping the gut microbiota using gene-specific gene expression studies and RNAi knockdown functional analyses. Candidate genes will also be tested for their implications in influencing the gut microbiota in fied mosquitoes collected in dengue endemic areas of Puerto Rico. This work will provide important insight into the molecular factors and mechanisms that shape the mosquito gut microbiota and contribute to its variability, which has been shown to influence vector competence. This information will advance our knowledge of mosquito gut immunity and will have important implications for the development of novel disease control strategies based on manipulation of the mosquito gut microbiota.