Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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Caspofungin enhances antifungal immunity in human blood through transcriptional reprogramming

ABSTRACT: This study investigates how the echinocandin antifungal drug caspofungin (CAS) modulates early immune responses during Candida albicans bloodstream infection. Echinocandin-susceptible (ECHS/FKSwt) and echinocandin-resistant (ECHR/FKSmut) C. albicans candidemia strains were used to determine whether CAS-induced host transcriptional changes depend on fungal susceptibility. Because immune modulation by CAS appears highly context dependent, we used an ex vivo human whole-blood system to capture physiologically relevant immune-pathogen interactions. Fresh human whole blood from healthy donors was infected ex vivo with either an echinocandin-susceptible (ECHS/FKSwt) or echinocandin-resistant (ECHR/FKSmut) C. albicans isolate. Samples were treated with CAS or left untreated as controls. After a defined incubation period, leukocyte RNA was isolated. Human mRNA sequencing was performed by Novogene (Novogene GmbH, Munich). RNA quality was assessed prior to library preparation, and globin mRNA was removed using GlobinClear. Sequencing was performed on an Illumina platform using 150 bp paired-end reads. Raw sequencing data underwent standard quality control, including removal of adapter sequences and low-quality reads. Clean reads were aligned to the human reference genome, and gene expression was quantified. Differential expression analyses were performed to identify host transcriptional responses to CAS during infections with susceptible versus resistant C. albicans strains.

INSTRUMENT(S): Illumina NovaSeq 6000

ORGANISM(S): Homo sapiens

SUBMITTER: Sascha Schäuble

PROVIDER: E-MTAB-16334 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Aspergillus fumigatus can cause Invasive Pulmonary Aspergillosis (IPA), the most severe form of Aspergillus-related infections. Fungicidal azoles and fungistatic caspofungin (CAS) are the first- and second-line therapies, respectively, used to treat IPA. However, azole-resistance is on the rise and new therapeutic alternatives need to be implemented. Recently, we showed that treatment of A. fumigatus with CAS or micafungin, but not voriconazole, induces production of the oxylipin 5,8-diHODE by the fungal oxygenase PpoA. Here we investigated the influence of ppo genes, that encode fatty acid oxygenases responsible for oxylipin biosynthesis, on CAS tolerance. The ppo mutants showed a very complex pattern of CAS susceptibility. PpoA and PpoC influence on CAS tolerance is mediated by MpkA phosphorylation and protein kinase A (PKA) activity. RNAseq transcriptional profiling and label-free quantitative proteomics (spectral counts) of the ppoA, and ppoC mutants showed that differentially expressed genes and proteins are related to secondary metabolites and carbohydrate metabolism. We tested several mutants of these differentially expressed genes encoding transcription factors and signal transduction proteins and some of them are more susceptible to CAS. We also characterized two clinical isolates, CNM7555 and IFM41607, that have decreased and increased susceptibility to CAS. CNM7555 does not have increased oxylipin production in the presence of CAS while the oxylipin induction upon CAS exposure is increased in the IFM41607, suggesting oxylipins are not the single mechanism involved in CAS tolerance in these isolates. Upon CAS exposure, CNM7555 has increased MpkA phosphorylation and PKA activity than IFM41607. Modulation of secondary metabolite production and carbohydrate metabolism is also essential in these clinical isolates for acquiring CAS tolerance. Our results reveal different aspects and genetic determinants involved in A. fumigatus CAS tolerance.

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Caspofungin enhances antifungal immunity in human blood through transcriptional reprogramming

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