The dynamics of Cryptococcus neoformans cell and transcriptional remodeling during infection

Purpose: Investigate the dynamics of cryptococcal cell and transcriptional remodeling during pulmonary infection in a murine model Methods: Dual-RNA sequencing Sample preparation - C57BL/6 mice were infected intratracheally with the H99 strain to analyze the transcriptome. After 6h or 240h of infection, BALF obtained from 6 mice of each condition were pooled together, generating two replicates of 6h and 240h (10d). BALF was centrifuged at 1200 x g and resuspended in 1mL TRIzol Reagent (Invitrogen Life Technologies) for RNA isolation, according to the manufactures' protocol. RNA from H99 yeasts grown in YPD was used as control. Results: Dual RNA-Seq analysis of C. neoformans and mice revealed molecular signatures of early and late events of infection We analyzed the up and downregulated genes after 6h and 10 days of infection, compared to the inoculated yeast (YPD x 6h and YPD x 10d) and the differentially expressed genes (DEGs) between 6h and 10 days of infection (designated BAL6h x BAL10d). These analyses revealed molecular signatures unique to early and late stages of infection. Conclusions: Our findings support a scenario in which the fungus initially deploys mechanisms aimed at survival, proliferation and tissue invasion. Later, the fungus may focus on mechanisms aimed at resistance to the immune response and colonization, that are crucial for disease development. Overall, our study provides new insights into understanding the biology of C. neoformans during infection and generating perspectives for the survey for new therapeutic and diagnostic targets for cryptococcosis. Overall design: C57BL/6 mice were infected intratracheally with the H99 strain to analyze the transcriptome. After 6h or 240h of infection, BALF obtained from 6 mice of each condition were pooled together, generating two replicates of 6h and 240h (10d).