Microbial interplay between ZIKA virus and the native microbiome in mosquitoes

ZIKA 病毒与蚊子体内微生物组之间的微生物相互作用

• 批准号: 9726225
• 负责人: Grant Leslie Hughes
• 金额: $ 13.5万
• 依托单位: TMLIVERPOOL SCHOOL OF TROPICAL MEDICINE
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-01 至 2020-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9726225
• 关键词: Aedes; Affect; African; Alphavirus; Antibiotics; Antiviral Agents; Arbovirus Infections; Arboviruses; Area; Arthropods; Asians; Attention; Bacteria; Biological Assay; Biology; Chikungunya virus; Competence; Complex; Culicidae; Data; Dengue Virus; Disease; Flavivirus; Foundations; Future; Generations; Gnotobiotic; High-Throughput Nucleotide Sequencing; Individual; Inherited; Invaded; Investigation; Investigative Reports; Knowledge; Laboratories; Mediating; Medical; Microbe; Modeling; Mosquito-borne infectious disease; Oral; Phenotype; Property; Role; Seminal; Shapes; Supplementation; Techniques; Technology; Variant; Viral; Viral Physiology; Virus Diseases; West Nile virus; Work; Yellow fever virus; ZIKV infection; Zika Virus; arbovirus disease; bacteriome; base; experimental study; fungus; gut microbes; insight; microbial; microbiome; microbiome composition; microbiota; microorganism; microorganism interaction; mycobiome; novel; offspring; pathogen; symbiont; vector; viral transmission

Project summary. The native microbiota of mosquitoes profoundly influences many aspects of mosquito biology, including the ability of mosquitoes to transmit arboviral pathogens. Perturbation of the mosquito microbiome, either by supplementation of antibiotics or by oral inoculation of bacterial isolates into the mosquito gut can interfere with flaviruses such as Dengue virus (DENV) and West Nile Virus as well as alphaviruses like Chikungyunya (CHIKV). Most studies indicate that gut microbes are antagonistic to pathogens, but particular bacteria facilitate viral infection in mosquitoes. Additionally, we are beginning to appreciate that arbovirus infection in mosquitoes can alter the bacterial microbiome. Taken together, it is clear that there is complex interplay, either by direct or indirect mechanisms, between native gut microbes of mosquitoes and invading arbovirus, and these interactions have a profound effect on vector competence. Here we will characterize the interplay between gut microbes of Aedes mosquitoes and Zika virus to determine how these interactions influence ZIKV vector competence in mosquitoes. Our work will focus on both bacterial and fungal taxa of Aedes aegypti and Aedes albopictus. While it is emerging that bacterial symbionts alter vector competence, we have a poor understanding of the role of fungal microbes on mosquito biology and vector competence. Our work will exploit high throughput sequencing technologies to obtain a culture-independent quantitative characterization of the bacterial and fungal microbiome. Importantly, experiments will use both lab-reared and field-collected mosquitoes as our appreciation of the role of the microbiome of on vector competence of mosquitoes in the field is poorly understood. In specific aim 1, we will identify bacterial and fungal taxa that alter vector competence by high throughput sequencing comparing perturbed and conventionally reared mosquitoes. Using field-collected mosquitoes, ZIKV vector competence will be correlated to microbial abundance and composition. As the composition of the microbiome is variable, the presence or abundance of specific symbionts may account for variation in arbovirus vector competence. ZIKV vector competence assays will be completed on gnotobiotic mosquito lines that have had isolates reinfected, functionally validating candidate taxa identified from our sequencing experiments. In specific aim 2, we will examine how ZIKV infection alters the bacterial and fungal microbiome using high throughput sequence. We will determine if microbes that ZIKV inhibits have anti-viral properties and examine if ZIKV-mediated modulation of the microbiome is transferred between generations. Lastly, we will determine if such transgeneration alteration of the micobiome affects vector competence of subsequent generations to ZIKV. This work will provide insights in microbial interactions within mosquito, a poorly understudied but important area of vector biology that has promising applications to control ZIKV and other mosquito-borne viral disease. This project will lay the foundation for future work in the Hughes laboratory, which is to develop applied novel microbial-based strategies suitable for simultaneously controlling multiple arboviruses such as ZIKV, DENV, CHIKV, and yellow fever virus.

项目概要。 蚊子的原生微生物群深刻影响蚊子生物学的许多方面，包括 蚊子传播虫媒病毒病原体。通过补充以下物质来扰乱蚊子微生物组 抗生素或将细菌分离株口服接种到蚊子肠道中可以干扰黄病毒，例如登革热 病毒（DENV）和西尼罗河病毒以及基孔肯亚病毒（CHIKV）等甲病毒。大多数研究表明肠道 微生物对病原体具有拮抗作用，但某些细菌会促进蚊子的病毒感染。此外，我们还 开始认识到蚊子中的虫媒病毒感染可以改变细菌微生物组。综合起来就是 清楚地表明，蚊子的原生肠道微生物之间存在着复杂的相互作用，无论是通过直接还是间接的机制 和入侵的虫媒病毒，这些相互作用对载体的能力产生深远的影响。在这里我们将描述 伊蚊和寨卡病毒肠道微生物之间的相互作用，以确定这些相互作用如何影响寨卡病毒 蚊子的媒介能力。我们的工作将重点关注埃及伊蚊和伊蚊的细菌和真菌类群 白纹伊蚊。虽然细菌共生体改变载体能力的现象正在出现，但我们对细菌共生体改变载体能力的了解还很有限。 真菌微生物对蚊子生物学和媒介能力的作用。我们的工作将利用高通量测序 获得细菌和真菌微生物组的独立于培养物的定量表征的技术。 重要的是，实验将使用实验室饲养的和现场收集的蚊子作为我们对蚊子作用的认识。 人们对野外蚊子媒介能力的微生物组知之甚少。在具体目标 1 中，我们将确定 通过高通量测序改变载体能力的细菌和真菌类群，比较扰动和 传统饲养的蚊子。使用现场采集的蚊子，ZIKV 载体能力将与 微生物丰度和组成。由于微生物组的组成是可变的，微生物组的存在或丰度 特定的共生体可能是虫媒病毒载体能力差异的原因。 ZIKV 载体能力检测将 在已分离出再感染的无菌蚊子品系上完成，对已识别的候选类群进行功能验证 来自我们的测序实验。在具体目标 2 中，我们将研究 ZIKV 感染如何改变细菌和真菌 使用高通量序列的微生物组。我们将确定 ZIKV 抑制的微生物是否具有抗病毒特性 并检查 ZIKV 介导的微生物组调节是否在代际间转移。最后，我们将 确定微生物组的这种转基因改变是否会影响后代的载体能力 寨卡病毒。这项工作将为蚊子体内的微生物相互作用提供见解，这是一个尚未得到充分研究但很重要的领域 载体生物学在控制 ZIKV 和其他蚊媒病毒性疾病方面具有广阔的应用前景。这个项目 将为休斯实验室未来的工作奠定基础，即开发基于微生物的新型应用 适用于同时控制多种虫媒病毒（例如 ZIKV、DENV、CHIKV 和黄热病）的策略 病毒。