In recent years, dengue fever has emerged as one of the most serious vector-borne viral diseases. The molecular biology of dengue virus propagation in the mosquito vector is largely unknown, but the genome sequence, reverse genetic and functional genomics tools as well as high throughput screening methodologies that are now available provide a means of dissecting the mosquito's interactions with the virus in greater detail. In this proposal, we will apply a highly innovative interdisciplinary approach to discover and develop small molecules (compounds) that can render the Aedes aegypti vector mosquito resistant to the virus and thereby incapable of transmitting disease. A previous collaboration between the groups of Drs. Garcia-Blanco and Dimopoulos identified over 300 potential dengue virus host and restriction factors through a high throughput RNAi screen in a Drosophila cell line. We will now expand on these findings and identify homologues of these factors in the Aedes mosquito in specific Aim 1.
In Aim 2 we will assess the anti-dengue potential of selected factors in the Aedes cell line and mosquito to select those that are most suitable for small molecule discovery. We will then use a powerful established yeast-based platform to identify compounds that can repress dengue virus host factors, and an established insect cell line -based system to identify compounds that can activate expression of dengue virus restriction factors in Aim 3.
In Aim 4 we will test a selected set of these compounds for anti-dengue activity in Aedes cell lines and adult female mosquitoes through different routes of exposure. We will finally investigate the potential influence of selected compounds on mosquito fitness parameters and assess their anti-dengue activity in field -derived mosquito strains in Aim 5. The completion of this highly innovative work will lead to a transition into translational studis on the delivery of potent anti-dengue compounds to natural dengue vector populations, as a prelude towards the development of a novel dengue control strategy.

Public Health Relevance

Despite the dramatic surge in dengue prevalence, surprisingly little is known about the molecular biology and genetic basis of the virus interactions with the mosquito vectors that enable transmission. This research proposal aims at the discovery and development of small molecules (compounds) that can target dengue virus host and restriction factors of the Aedes aegypti mosquito, and thereby renders it incapable of transmitting the disease. The knowledge generated from this study will furthermore contribute to the development of control methods for dengue and other viruses such as yellow fever and West Nile viruses.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI101431-02
Application #
8461517
Study Section
Vector Biology Study Section (VB)
Program Officer
Costero, Adriana
Project Start
2012-05-01
Project End
2017-04-30
Budget Start
2013-05-01
Budget End
2014-04-30
Support Year
2
Fiscal Year
2013
Total Cost
$723,643
Indirect Cost
$106,290
Name
Johns Hopkins University
Department
Microbiology/Immun/Virology
Type
Schools of Public Health
DUNS #
001910777
City
Baltimore
State
MD
Country
United States
Zip Code
21218
Krishnan, Manoj N; Garcia-Blanco, Mariano A (2014) Targeting host factors to treat West Nile and dengue viral infections. Viruses 6:683-708
Jupatanakul, Natapong; Sim, Shuzhen; Dimopoulos, George (2014) Aedes aegypti ML and Niemann-Pick type C family members are agonists of dengue virus infection. Dev Comp Immunol 43:1-9
Sim, Shuzhen; Jupatanakul, Natapong; Dimopoulos, George (2014) Mosquito immunity against arboviruses. Viruses 6:4479-504
Kang, Seokyoung; Shields, Alicia R; Jupatanakul, Natapong et al. (2014) Suppressing dengue-2 infection by chemical inhibition of Aedes aegypti host factors. PLoS Negl Trop Dis 8:e3084
Barrows, Nicholas J; Jamison, Sharon F; Bradrick, Shelton S et al. (2014) Functional genomics approach for the identification of human host factors supporting dengue viral propagation. Methods Mol Biol 1138:285-99
Jupatanakul, Natapong; Sim, Shuzhen; Dimopoulos, George (2014) The insect microbiome modulates vector competence for arboviruses. Viruses 6:4294-313
Sim, Shuzhen; Jupatanakul, Natapong; Ramirez, Jose L et al. (2013) Transcriptomic profiling of diverse Aedes aegypti strains reveals increased basal-level immune activation in dengue virus-refractory populations and identifies novel virus-vector molecular interactions. PLoS Negl Trop Dis 7:e2295