Multi-omics to probe Acanthamoeba extracellular vesicles

Free-living amoebae of the genus Acanthamoeba are protists widely distributed in the environment, including water, soil, and air, as well as in human-made habitats such as swimming pools and air conditioning systems. These amoebae can act as opportunistic pathogens when they come into accidental contact with humans, leading to various severe diseases. The most common one is Acanthamoeba keratitis (AK), a painful, sight-threatening, and difficult-to-treat ocular infection, particularly common in contact lens wearers. This is because these amoebae often contaminate contact lens cases, proliferate to high densities, attach to contact lenses, and infect the cornea upon contact with the eye. The process of infection depends on the number and virulence of the respective amoeba, the host`s susceptibility, and environmental conditions. The parasite`s ability to initiate cell-to-cell contact and bind to epithelial cells is considered the basis of infection. However, as established for many pathogens, cell-to-cell communication without direct contact, by the release of lipid- enclosed bilayers termed extracellular vesicles (EVs), may also play a significant role during infection. Despite considerable progress in recent years, the pathogenesis of Acanthamoeba infection is still not fully understood. In recent years, comparative and integrative omics have become important tools for detailed molecular analyses. In this project, we aim to comprehensively explore the proteomic, genomic, and transcriptomic profiles of EVs secreted by Acanthamoeba upon contact with human corneal epithelial cells and to enable their molecular characterization. We will further complement these approaches by imaging techniques on the light and electron microscopical level. The data generated will provide fundamental new insights at the molecular level into the secretory mechanisms used by Acanthamoeba during AK infection, including their effects and structural integrity. The findings will significantly improve our understanding of the pathogenic cascade in AK and may also aid in the development of future intervention strategies.