Project name: Characterization of Entamoeba histolytica adapted to auranofin by combined transcriptomics and redox proteomics

Description: Purpose: Transcriptome characterization of Entamoeba histolytica trophozoites adapted to AuranofinTotal RNA was extracted from control trophozoites (WT) and AFAT using the TRI reagent kit, according to the manufacturer instructions (Sigma-Aldrich USA). 6 RNAseq libraries were produced according to manufacturer protocol (NEBNext UltraII Directional RNA Library Prep Kit for Illumina, cat no. E7760) using 800ng total RNA. mRNAs pull-up was performed using Magnetic Isolation Module (NEB, cat no. E7490). All libraries were mixed into a single tube with equal molarity. The RNAseq data was generated on Illumina NextSeq500, 75 single-end read, high output mode (Illumina, cat no. 200249) The number of reads per gene was counted using Htseq-count (v0.9.1)Results: Transcriptomics of Entamoeba histolytica trophozoites that were adapted to 2 uM Auranofin revealed an upregulation of genes encoding cytoskeletal proteins, dehydrogenases and guanyl-nucleotide exchange factors.Conclusions: Adaptation to Auranofin comes with a fitness cost for E.histolytica that includes a decreased growth rate and virulence and sensitivity to Oxidative stress, Nitrosative stress and to Metronidazole. Overexpression of genes whose products are sensitive to Auranofin-mediated oxidation may represent an important step in the adaptation process to Auranofin and EhTrxR does not seem to be central for this process.Overall design: Entamoeba histolytica mRNA profiles of WT and Auranofin adapted (2uM) trophozoites

Data type: Transcriptome or Gene expression

Sample scope: Multiisolate

Relevance: Medical

Last updated: 2021-06-20