Investigating the Roles of Protein Receptors GPR4 and GPR5 in the Hypervirulence in the Clinical Isolate UgCl223 of Cryptococcus Neoformans
The pathogenic yeast, Cryptococcus neoformans, is the most common cause of fungal meningitis. C. neoformans is commonly encountered via inhalation of the saprophyte in the environment. Infection typically spreads from the lung to the brain by way of the bloodstream, once in the lungs the latent infection reactivates then disseminates, eventually crossing the blood brain barrier and resulting in cryptococcal meningitis. Cryptococcal meningitis is especially dangerous to immunocompromised populations and is associated with HIV/AIDS. Previous studies showed the protein receptors Gpr4 and Gpr5 are required for titan cell formation, a stress response that is hypothesized to be closely associated with the latent infection. To further study the role of GPR4 and GPR5 in a latent infection model, we simultaneously deleted both genes in the latent clinical isolate UgCl223 using the Crispr/Cas9 system. This was the first time simultaneous deletion was used to create a C. neoformans mutant strain. The gpr4Δ gpr5Δ deletion mutant is being tested for titan cell production in vitro and in vivo and will be compared to the wild type strain, as well as the double deletion in the lab reference strain KN99?. The data obtained will provide additional evidence of the relationship between the receptors Gpr4 and Gpr5 in C. neoformans and the formation of titan cells during latent infection.