Functional Genomic Analysis of Candida glabrata-Macrophage Interaction: Role of Chromatin Remodeling in Virulence

Abstract

Fungal septicemia is an increasingly common complication of immunocompromised patients worldwide. Candida species are the leading cause of invasive mycoses with Candida glabrata being the second most frequently isolated Candida species from Intensive Care Unit patients. Despite its clinical importance, very little is known about the mechanisms that C. glabrata employs to survive the antimicrobial and immune response of the mammalian host. Here, to decipher the interaction of C. glabrata with the host immune cells, we have screened a library of 18,350 C. glabrata Tn 7 insertion mutants for reduced survival in human THP-1 macrophages via signature-tagged mutagenesis approach. A total of 56 genes, belonging to diverse biological processes including chromatin organization and golgi vesicle transport, were identified which are required for survival and/or replication of C. glabrata in macrophages. We report for the first time that C. glabrata wild-type cells respond to the intracellular milieu of macrophage by modifying their chromatin structure and chromatin resistance to micrococcal nuclease digestion, altered epigenetic signature, decreased protein acetylation and increased cellular lysine deacetylase activity are the hall-marks of macrophage-internalized C. glabrata cells. Consistent with this, mutants defective in chromatin organization (Cgrsc3-aΔ, Cgrsc3-bΔ, Cgrsc3-aΔbΔ, Cgrtt109Δ) and DNA damage repair ( Cgrtt107Δ, Cgsgs1Δ) showed attenuated virulence in the murine model of disseminated candidiasis. Further, genome-wide transcriptional profiling analysis on THP-1 macrophage-internalized yeasts revealed deregulation of energy metabolism in Cgrsc3-aΔ and Cgrtt109Δ mutants. Collectively, our findings establish chromatin remodeling as a central regulator of survival strategies which facilitates a reprogramming of cellular energy metabolism in macrophage-internalized C. glabrata cells and provide protection against DNA damage.

Author Summary

Hospital-acquired fungal infections pose a colossal health and economic challenge. Candida species are the leading cause of disseminated fungal infections and rank fourth among the most common nosocomial pathogens. C. glabrata, an emerging opportunistic fungal pathogen, is the second most frequently isolated Candida species after C. albicans from Intensive Care Unit patients world-wide. Limited information is available on the unique strategies that C. glabrata employs to evade and replicate in host phagocytic cells since it lacks the key virulence traits of C. albicans including hyphal formation and secreted proteolytic activity. In the current study, we have identified a total of 56 genes, via a functional genomics approach, which are required for survival and/or replication of C. glabrata in human macrophages. Our data demonstrates an essential role for chromatin remodeling in the intracellular survival of C. glabrata with ingested C. glabrata cells displaying transcriptionally active chromatin in early-phase, compact, closed chromatin in mid-stage, and open chromatin in the late-stage of macrophage internalization. Our findings identify novel fungal virulence determinants and potentially implicate epigenetic changes in the metabolic adaptation of fungal cells to the nutrient-poor host environment and the survival against oxidative stress-induced DNA damage.