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How Do Arboviruses Escape the Mosquito Midgut?- Analysis of a Novel Mechanism

虫媒病毒如何逃离蚊子中肠？-新机制的分析

基本信息

批准号：
10289718
负责人：
Alexander W E Franz
金额：
$ 38.27万
依托单位：
UNIVERSITY OF MISSOURI-COLUMBIA
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-11-03 至 2024-10-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10289718
关键词：
ADAMTS Aedes Affect Applications Grants Arboviruses Basal lamina Biochemical CRISPR/Cas technology Cathepsin L Chikungunya virus Collagen Type IV Communities Complex Culicidae Data Dengue Dengue Virus Development Digestion Enzymes Epithelial Cells Eukaryota Exclusion Extracellular Matrix Gene Expression Genes Genetic Genetic Markers Human Imaging technology In Situ Individual Infection Ingestion Interstitial Collagenase Ions Laminin Matrix Metalloproteinases Mayaro virus Metalloproteases Methodology Midgut Modification Molecular Nature Oral Organ Organism Outcome Peptide Hydrolases Peptidoglycan Phenotype Population Post-Translational Protein Processing Process Proteins Proteomics RNA interference screen Research Research Personnel Resolution Saline Salivary Glands Scanning Electron Microscopy Secondary to Serine Stretching Structure Testing Thick Time Tissue Inhibitor of Metalloproteinases Tissues Transgenic Organisms Variant Viral Virion Virus Virus Diseases ZIKA Zika Virus base chikungunya collagenase detection assay experimental study interest knockout gene molecular marker monolayer mosquito-borne novel novel strategies novel virus overexpression polyclonal antibody proteinase In three dimensional structure tool transcriptome sequencing vector vector competence vector mosquito viral transmission

项目摘要

Title: How do arboviruses escape the mosquito midgut? - Analysis of a novel mechanism Project Summary. Following ingestion of a viremic bloodmeal from a vertebrate host, an arbovirus enters the midgut lumen of a mosquito vector (such as Aedes aegypti) along with the bloodmeal. The virus then needs to enter and infect the midgut epithelial cells of the midgut before the virus disseminates from the midgut to secondary tissues including the salivary glands. Once these are infected, the mosquito transmits the virus to another vertebrate host during probing. The molecular nature of the exit mechanism of the virus from the midgut, has been unresolved so far. In this grant application, we will reveal this mechanism and its genetic background using a multifaceted portfolio of state-of-the-art methodologies. Our previous studies demonstrated that during bloodmeal digestion, chikungunya virus (CHIKV) exits the mosquito midgut via the basal lamina (BL) surrounding the organ. The BL predominantly consists of collagen IV and laminin and its typical pore size exclusion limit appears to be too small for virions to pass through. However, during bloodmeal digestion, the BL alters its structure as midgut-associated collagen IV becomes greatly diminished. We hypothesized that it is this structural change in the BL that allows virions to exit the midgut during bloodmeal digestion. In eukaryotes, known proteinases that modify the extracellular matrix including the BL are matrix metalloproteinases, ADAM/ADAMTS, and serine collagenases. We hypothesize that these classes of proteinases, which are also present in mosquitoes, are responsible for midgut BL modification during bloodmeal digestion. To test these two hypotheses, we propose the following three Specific Aims for this application: 1.) Analyze BL degradation/remodeling and CHIKV dissemination in Ae. aegypti by ultrastructural studies and proteomic analysis; 2.) Identify proteinases that are involved in the BL modification process during bloodmeal digestion and CHIKV dissemination; 3.) Assess whether other viruses (such as Zika, dengue, and Mayaro viruses) utilize the same midgut escape mechanism as observed for CHIKV and how transgenic manipulation of proteinase-of-interest expression affects viral midgut escape in mosquitoes. Completion of these Specific Aims will provide a comprehensive picture explaining the mechanism of the midgut escape barrier, which key enzymes are involved in the mechanism, and whether the mechanism elucidated for CHIKV is the paradigm for other arboviruses. Our results will be critical for developing novel control strategies aimed at manipulating arbovirus midgut escape in mosquitoes. Our findings will also provide the research community with the possibility to develop novel genetic markers for vector competence based on midgut escape.

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