A functional and comparative approach for the study of Wolbachia-mediated virus blocking in Aedes aegypti and Aedes albopictus

研究沃尔巴克氏体介导的埃及伊蚊和白纹伊蚊病毒阻断的功能和比较方法

• 批准号: 10455257
• 负责人: Elizabeth Ann McGraw
• 金额: $ 49.18万
• 依托单位: PENNSYLVANIA STATE UNIVERSITY, THE
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-04 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10455257
• 关键词: Aedes; Antibodies; Antiviral Agents; Asian Americans; Australia; Bacteria; Biocontrols; Brazil; Candidate Disease Gene; Cell Adhesion; Cellular Stress; Chikungunya virus; Colombia; Communities; Culicidae; Data; Dengue; Dengue Fever; Dengue Infection; Dengue Virus; Development; Drug or chemical Tissue Distribution; Effectiveness; Future; Gardenal; Genes; Genetic; Genetic Transcription; Genetic Variation; Genome; Genotype; Geography; Goals; Government; Health; Human; Immune; Incidence; Indonesia; Infection; Inherited; Insecta; Knowledge; Laboratories; Location; Malaysia; Mediating; Mexico; Modeling; Nature; Neurons; Outcome; Population; Population Heterogeneity; Privatization; Process; Proteins; RNA Interference; Reactive Oxygen Species; Reporter; Research; Resistance; Resources; Risk; Role; Singapore; Sister; Site; Testing; Time; Tissues; Urbanization; Vaccines; Variant; Vietnam; Viral; Virus; Virus Replication; Wolbachia; Yellow Fever; Yellow fever virus; ZIKA; Zika Virus; chikungunya; climate change; comparative; density; design; endosymbiont; experience; experimental study; geographically distant; human disease; improved; mosquito reproduction; novel; programs; regional difference; response; symbiont; theories; trait; transmission process; vector; vector competence; vector control; vector mosquito; viral transmission; virus genetics; virus host interaction; weapons

PROJECT SUMMARY/ABSTRACT The global range of the mosquito, Aedes aegypti continues to expand due to a changing climate and increasing urbanization. As Ae. aegypti is the primary vector of dengue, Zika, yellow fever and chikungunya viruses, these range shifts also increase the risk of human disease. With few effective vaccines or antiviral drugs, vector control remains the best weapon against mosquitoes. Recently, the release of an insect endosymbiont, Wolbachia, into mosquito populations has emerged as a novel biocontrol strategy. Maternally inherited and with inbuilt genetic drive, Wolbachia tends to spread through insect populations. Inside the mosquito, the bacterium limits the replication of viruses, a trait called ‘viral blocking’. Following release of Wolbachia into wild populations, the viral blocking effect is leading to substantial reductions in the incidence of dengue fever in humans. One concern for the long- term efficacy of Wolbachia is the emergence of resistance in mosquitoes or viruses. To design strategies to counter this likely scenario, we need to understand the genetic basis of viral blocking. While many theories have been proposed, none have been entirely satisfying. Recently, the McGraw lab used a selection experiment to create Ae. aegypti lines carrying Wolbachia with differences in blocking. In sequencing those lines, her group identified a set of novel candidate loci and a new set of hypotheses for the basis of blocking. This study also demonstrated that the strength of blocking is likely to vary across diverse mosquito genetic backgrounds. Here we propose to provide functional testing for the candidate blocking genes involved with cell adhesion, transcriptional pausing and neuronal function, for both dengue and chikungunya viruses using RNA silencing. In these experiments we will also contrast Aedes albopictus, that has long harbored a natural infection of Wolbachia, against Ae. aegypti with a recently artificially introduced Wolbachia. The nature of blocking in Ae. albopictus is a likely harbinger of the future of blocking in the artificially infected species. We will also carry out large common garden experiments to dissect the relative contribution of regional differences in mosquito and virus diversity to the strength of blocking. This project will produce novel resources for the community including a new Wolbachia infected Ae. aegypti line, a set of antibodies against key blocking association proteins and data on the efficacy of improved fluorescent reporter viruses for the study of vector competence. More broadly, this research will help to test a revised model for Wolbachia-mediated viral blocking – knowledge fundamental for designing strategies to prolong Wolbachia’s efficacy in the field. It will also reveal the robustness of blocking to mosquito and viral genetic diversity, that will inform strategies for future Wolbachia strain deployment globally.

项目概要/摘要 由于气候变化和气候变化，埃及伊蚊的全球分布范围持续扩大。 由于埃及伊蚊是登革热、寨卡病毒、黄热病的主要传播媒介。 与基孔肯雅病毒相比，这些范围的变化也增加了人类患病的风险，但几乎没有什么有效的方法。 疫苗或抗病毒药物，媒介控制仍然是对抗蚊子的最佳武器。 将昆虫内共生体沃尔巴克氏体释放到蚊子种群中已成为一种新颖的方法 生物防治策略是母系遗传且具有内在的基因驱动，沃尔巴克氏体倾向于传播。 通过昆虫种群，细菌限制了病毒的复制，这是一种特征。 沃尔巴克氏体释放到野生种群后，病毒阻断效果被称为“病毒阻断”。 导致人类登革热发病率大幅下降，这是长期关注的问题之一。 沃尔巴克氏体的术语功效是对蚊子或病毒产生抗药性。 为了应对这种可能的情况，我们需要了解病毒阻断的遗传基础。 尽管已经提出了许多理论，但最近没有一个理论完全令人满意。 实验室通过选择实验创建了携带沃尔巴克氏体的埃及伊蚊品系，其差异在于： 在对这些细胞系进行测序时，她的团队确定了一组新的候选基因座和一组新的基因座。 阻塞基础的假设本研究还证明了阻塞的强度是可能的。 在此我们建议提供功能测试。 候选阻断基因涉及细胞粘附、转录暂停和神经 在这些实验中，我们将使用 RNA 沉默来针对登革热和基孔肯雅病毒发挥作用。 还对比了长期携带沃尔巴克氏体自然感染的白纹伊蚊对抗伊蚊的情况。 埃及伊蚊与最近人工引入的沃尔巴克氏体的阻断性质是白纹伊蚊。 可能预示着未来我们还将开展大规模的人工感染物种封锁。 常见的花园实验来剖析蚊子和蚊子的区域差异的相对贡献 该项目将为社区提供新颖的资源。 包括一种新的沃尔巴克氏体感染的埃及伊蚊系，一组针对关键阻断关联的抗体 蛋白质和改进的荧光报告病毒在载体研究中的功效数据 更广泛地说，这项研究将有助于测试沃尔巴克氏体介导的病毒的修订模型。 阻断——设计策略以延长沃尔巴克氏体在该领域的功效的基础知识。 它还将揭示对蚊子和病毒遗传多样性的阻断的稳健性，这将告知 未来沃尔巴克氏菌菌株在全球部署的策略。