A Genomic and Proteomic Approach for the Evaluation of Exosomes from Toxocara Excretory Secretory (TES) Products from Toxocara canis and Toxocara cati Larval Cultures
Principal Investigator: Manigandan Lejeune
DESCRIPTION (provided by applicant):
Toxocara is a genus of nematodes, which infects a variety of hosts, principally dogs and cats, with potential zoonotic risks to humans. Toxocara spp. larvae are capable of migrating throughout the host tissues, eliciting eosinophilic and granulomatous reactions, while surviving for extended periods of time, unchanged, in the host. It is postulated that larvae are capable of altering the host’s immune response through the elaboration of excretory-secretory products, containing both proteins and extracellular vesicles. The study of extracellular vesicles, particularly exosomes, has increased exponentially in recent years, largely due to their potential use as a diagnostic tool, and in molecular therapy. Here, we will isolate exosomes acquired from larval cultures of Toxocara canis and Toxocara cati containing Toxocara excretory-secretory products (TES). We will analyze isolated exosomes and remaining TES proteins and nucleic acids through RNA sequencing, mass spectrometry, western blotting, and confocal microscopic visualization of fluorescently tagged exosomes. We will also analyze the effects of TES on RNA expression of human and mouse cells by performing RNA sequencing following exposure to exosomes and TES proteins. The RNA sequences with significant changes in expression will be then annotated to allow for interpretation of effects on cellular functions following exposure to TES.