Effects of Wolbachia and arboviruses on lipid metabolism in mosquitoes

The major aim will be to further understanding of lipid metabolism in response to arbovirus infection and presence of Wolbachia symbionts in Aedes mosquitoes. Arboviruses from three different families will be used to initially infect Aedes cell lines both with and without Wolbachia: Semliki forest virus (Alphavirus), Zika virus (Flavivirus) and bunyamwera virus (Orthobunyavirus). Lipidomics will be carried out to determine the overall global change in lipid metabolism in response to infection with Wolbachia. Cyclodextrins, a group of compounds used to study lipid turnover and dynamics, will be utilized alongside labelled cholesterol to understand spatially and temporally how different conditions effect lipid metabolism. A more fundamental understanding of spatial dynamics of lipids will be determined using Raman scattering spectroscopy. Raman is an information-rich vibrational technique that gives label-free biochemical information at the sub-cellular level. However, high-resolution imaging at a single-cell level can be a relatively slow process, making it challenging for imaging of larger cell populations and live-cell imaging. Stimulated Raman scattering (SRS) is a more sensitive and faster technique which can provide high resolution information on larger cell populations compared with normal Raman. Both Raman and SRS provide detailed biochemical information with sub-cellular resolution including determination of protein, lipids and nucleic acid location. In this project we will use a combination of Raman and SRS to determine lipid content & composition, spatial location and changes in response to infection. Chemometric analysis techniques will be applied to hyperspectral SRS imaging datasets to provide meaningful analysis on the changes in the lipid metabolism profile during and following infection. Once key lipid metabolites are determined the student will manipulate these metabolites using targeted drugs and knockdown / knockout systems to determine how they affect Wolbachia and arboviral infection.