Project description:With the rise in immunocompromised individuals and patients with immune-related comorbidities such as COVID-19, the rate of fungal infections is growing. This increase, along with the current plateau in anti-fungal drug development, has made understanding the pathogenesis and dissemination of these organisms more pertinent than ever. The mouse model of fungal infection, while informative on a basic science level, has severe limitations in terms of translation to the human disease. Here we present data supporting the implementation of the human cerebral organoid model, which is generated from human embryonic stem cells and accurately recapitulates relevant brain cell types and structures, to study fungal infection and dissemination to the central nervous system (CNS). This approach provides direct insight into the relevant pathogenesis of specific fungal organisms in human tissues where in vivo models are impossible. With this model system we assessed the specific brain tropisms and cellular effects of fungal pathogens that are known to cross the blood brain barrier such as Cryptococcus neoformans. We determined the effects this fungal pathogen has on the overall gross morphology, cellular architecture, and cytokine release of these model organoids. Furthermore, we demonstrated that C. neoformans can penetrate and invade the organoid tissue and remain present throughout the course of infection. These results demonstrate the utility of this new model to the field and highlight the potential for this system to elucidate fungal pathogenesis to new therapeutic strategies to prevent and treat the terminal stages of fungal diseases such as cryptococcal meningitis.
Project description:We measured protein translation (by ribosome profiling) and RNA levels (by polyA-enriched RNA-seq) in Cryptococcus neoformans strain H99 and Cryptococcus neoformans strain JEC21. This is the first transcriptome-wide map of translation in this species complex.
Project description:We investigated the effects of the hypoxia-mimetic CoCl2 on the gene expression of pathogenic fungus Cryptococcus neoformans. Keywords: compound treatment design
Project description:Light is a universal environmental signal perceived by many organisms, including the fungi in which light regulates both common and unique biological processes depending on the species. We conducted a whole-genome microarray analysis on the basidiomycete fungus Cryptococcus neoformans to identify light-regulated genes.
Project description:Discovering the Roles of the Casein Kinase 2 Complex in the Growth, Differentiation, Stress Responses, and Pathogenicity of Cryptococcus neoformans
Project description:Cryptococcal meningitis (CM) is a fatal compliation. Macrophages work as Trojan horses transferring Cryptococcus neoformans (C. neoformans) into the brain. Mechanisms for C. neoformans Trojan horses are largely elusive. In this study, we performed scRNA-Seq on immune cells infiltrated into the brain in a murine model of CM. Bioinformatics analysis reveals that phosphodiesterase 4B (PDE4B) ranks top in regulating macrophage Trojan horses. Melanin, a virulence factor for Cn, decreased PDE4B in macrophages. PDE4B inhibitor promoted the Cn Trojan horse across the blood-brain-barrier (BBB) in vitro and in vivo. As similar, PDE4B knockout increased fungi burden in the brain, which is, at least partially, rescued by macrophages depletion. In contrast, PDE4B activation diminished C. neoformans brain infection. Mechanistically, PDE4B inhibition increased CXCR4 and CCR7 on C. neoformans macrophage Trojan horses, a process regulated by the cAMP/PKA signaling pathway. Dexamethasone, commonly used to treat Pneumocystis infections in AIDS patients, significantly decreased PDE4B expression in macrophages. Overall, this study reveals that PDE4B plays a crucial role in C. neoformans macrophage Trojan horses and serves as a potential therapeutic target for CM.