Entamoeba histolytica
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PROVIDER: PRJNA151677 | ENA |
REPOSITORIES: ENA
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Project description:Purpose: Transcriptome characterization of Entamoeba histolytica trophozoites adapted to Auranofin Total 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.
2021-08-31 | GSE178520 | GEO
Project description:The ability of Entamoeba histolytica to phagocytose host cells correlates to observed virulence in vivo. To better understand the mechanism of phagocytosis we used paramagnetic beads coated with host ligand and sorted trophozoites based on phagocytic ability. Gene expression was then measured in both the sorted phagocytic and non-phagocytic populations using a custom Affymetrix chip for E. histolytica. Feed forward regulation of phagocytosis by Entamoeba histolytica. Infection and Immunity. PMID 23045476
2012-10-16 | GSE41604 | GEO
Project description:Differential expression was used to access gene differences after Entamoeba histolytica infection. Entamoeba histolytica is an important diarrheal pathogen worldwide, and induces apoptosis of the intestinal epithelium as part of its disease process. Regenerating (REG) 1 protein is anti-apoptotic. We investigated the involvement of REG 1 in E. histolytica colitis. Colonic biopsy samples were obtained from 8 subjects with acute E. histolytica colitis, and again 60 day later during convalescence. Gene expression in the human colon during acute and convalescent E. histolytica disease was evaluated using microarray and confirmed by polymerase chain reaction (PCR). REG 1 protein expression was evaluated with immunohistochemistry. The mechanism of REG 1 involvement in E. histolytica disease was subsequently investigated with a mouse model. REG 1A and REG 1B were the most upregulated genes in the human intestine in acute versus convalescent E. histolytica disease (p=0.003 and p=0.006 respectively). PCR confirmed the microarray results (p=<0.001 and p=0.001 respectively). Increased REG 1A and REG 1B protein expression was similarly observed by immunohistochemistry. REG 1 -/-mice were found to be significantly more susceptible to E. histolytica infection than wild type mice.
2011-05-24 | GSE23750 | GEO
Project description:Entamoeba histolytica is a protozoan parasite that causes colitis and liver abscesses. Several Entamoeba species and strains with differing levels of virulence have been identified. E. histolytica HM-1:IMSS is a virulent strain, E. histolytica Rahman is a nonvirulent strain, and Entamoeba dispar is a nonvirulent species. We used an E. histolytica DNA microarray consisting of 2,110 genes to assess the transcriptional differences between these species/strains with the goal of identifying genes whose expression correlated with a virulence phenotype. We found 415 genes expressed at lower levels in E. dispar and 32 genes with lower expression in E. histolytica Rahman than in E. histolytica HM-1:IMSS. Overall, 29 genes had decreased expression in both the nonvirulent species/strains than the virulent E. histolytica HM-1:IMSS. Interestingly, a number of genes with potential roles in stress response and virulence had decreased expression in either one or both nonvirulent Entamoeba species/strains. These included genes encoding Fe hydrogenase (9.m00419), peroxiredoxin (176.m00112), type A flavoprotein (6.m00467), lysozyme (6.m00454), sphingomyelinase C (29.m00231), and a hypothetical protein with homology to both a Plasmodium sporozoite threonine-asparagine-rich protein (STARP) and a streptococcal hemagglutinin (238.m00054). The function of these genes in Entamoeba and their specific roles in parasite virulence need to be determined. We also found that a number of the non-long-terminal-repeat retrotransposons (EhLINEs and EhSINEs), which have been shown to modulate gene expression and genomic evolution, had lower expression in the nonvirulent species/strains than in E. histolytica HM-1:IMSS. Our results, identifying expression profiles and patterns indicative of a virulence phenotype, may be useful in characterizing the transcriptional framework of virulence.
2006-01-03 | E-SMDB-3589 | biostudies-arrayexpress
Project description:Up until now, the existence of Dnmt2-mediated DNA methylation has mostly been supported by focal analyses in organisms that contain Dnmt2, but no Dnmt1 or Dnmt3 DNA methyltransferase. In these organisms, several independent studies have also provided support for a biologically important function of Dnmt2-dependent DNA methylation. For example, Dnmt2-dependent methylation in Entamoeba histolytica, the causative agent of amebic dysentery, has been connected to the parasite s virulence. However, global DNA methylation levels in Entamoeba have been found to be very low. In addition, no specific features, such as CpG-specificity and specificity for certain genetic subcompartments have been described. This distinguishes Dnmt2-dependent methylation patterns from all other known methylomes and has raised questions about the validity of the underlying results. We have used whole-genome bisulfite sequencing for an unbiased characterization of the Entamoeba histolytica methylome at single-base resolution in a E.histolytica strain HM-1:IMSS devoid of significant level of EhDnmt2 (Ehmeth) expression.
2011-08-10 | GSE31295 | GEO
Project description:Entamoeba histolytica clinical isolates from NCGM
| PRJDB8679 | ENA
Project description:Previously, we have shown that in Entamoeba histolytica, methionine γ-lyase (EhMGL) downregulation results in trifluoromethionine resistance. The transcriptional response, however, of this parasite to the drug is not known. In this study, we used microarray analysis to determine if additional genes are involved. The expression profile of 9,230 genes in wild type and trifluoromethionine resistant strains was compared. Episomal overexpression of EhBspA1 was performed to verify its role in trifluoromethionine resistance. The transcriptomes of trifluoromethionine resistant strain cultured with or without trifluoromethionine, EhMGL-gene silenced strain, metronidazole resistant strain, and wild type strain under cysteine-deprived conditions were compared to determine the specificity of the changes observed in trifluoromethionine resistant strain.
2011-11-16 | E-GEOD-32314 | biostudies-arrayexpress
Project description:In vitro studies have described metronidazole (MTZ) resistance and the potential mechanisms involved. Costs to fitness and adaptive responses associated with resistance, however, have not been investigated. In this study we generated an HM-1-derived strain resistant to 12 µM MTZ (MTZR). We examined its phenotypic and transcriptional profile to determine the consequences and mRNA level changes associated with MTZ resistance. The expression profile of 9,230 genes in wild-type and metronidazole-resistant strains was compared. Transcriptome analysis revealed 142 differentially expressed genes in MTZR. In contrast to other MTZ-resistant parasites, MTZR did not down-regulate pyruvate:ferredoxin oxidoreductase, but showed increased expression of genes for a hypothetical protein (HP1) and several iron-sulfur flavoproteins, and the downregulation of genes for leucin-rich proteins. Overexpression of HP1-HA did not confer MTZR level of resistance, but provided a slight advantage in cell survival. Fisher's exact test showed 24 significantly enriched GO terms in MTZR, and a 3-way comparison of modulated genes with MTZR cultured without MTZ and HM-1 cultured with MTZ showed that 88 genes were specific to MTZR. Wild-type and the MTZ-resistant strain MTZR were cultured either in the presence or absence of MTZ. 2 biological replicates per strain/condition.
2012-02-22 | E-GEOD-35990 | biostudies-arrayexpress
Project description:The ability of Entamoeba histolytica to phagocytose host cells correlates to observed virulence in vivo. To better understand the mechanism of phagocytosis we used paramagnetic beads coated with host ligand and sorted trophozoites based on phagocytic ability. Gene expression was then measured in both the sorted phagocytic and non-phagocytic populations using a custom Affymetrix chip for E. histolytica. Feed forward regulation of phagocytosis by Entamoeba histolytica. Infection and Immunity. PMID 23045476 M280 Streptavidin Dynabeads (Invitrogen) were labeled with 20ug/mL biotinylated Human C1q (Quidel). Entamoeba histolytica (strain HM1) were washed twice with PBS then resuspended with the labeled beads at a 10:1 ratio of beads to trophozoites. Samples were incubated at 37°C for 45 minutes, washed twice with agitation to remove adherent beads, then resuspended in MACS buffer. Samples were loaded into magnetic columns (Miltenyi Biotec) and trophozoites were seperated according to manufacturer's protocols. phagocytic vs. non-phagocytic Entamoeba histolytica populations
2012-10-16 | E-GEOD-41604 | biostudies-arrayexpress