Project description:This data is part of a pre-publication release. For information on the proper use of pre-publication data shared by the Wellcome Trust Sanger Institute (including details of any publication moratoria), please see http://www.sanger.ac.uk/datasharing/The drug Praziquantel is the most commonly used drug for parasitic flatworms. It is currently being increasingly used in mass drug administration programmes, raising concerns over whether resistance will develop. Although widely used, its mode of action was until very recently uncertain. This study will investigate the praziquantel mode of action and resistance by sequencing the transcriptomes of Dugesia japonica and different stages and strains of S. mansoni.
Project description:This SuperSeries is composed of the following subset Series: GSE21008: Linking toxicant physiological mode of action with induced gene expression changes in Caenorhabditis elegans: atrazine GSE21010: Linking toxicant physiological mode of action with induced gene expression changes in Caenorhabditis elegans: cadmium GSE21011: Linking toxicant physiological mode of action with induced gene expression changes in Caenorhabditis elegans: fluoranthene Refer to individual Series
Project description:The anti-mycobacterial activity of C17 diynes has been described previously, however, their mode of action remains unknown. Microarray techniques were used to explore the genetic regulation reponses of Mycobacterium smegmatis to treatment with the C17 diynes, falcarinol and panaxydol. Our analyses showed a distinct mode of action of the C17 diynes when compared with commonly used anti-mycobacterial drugs. In addition, geneset enrichment analysis, pathway enrichment analysis and PASS analysis revealed significant gene ontology terms, pathways and potential modes of action, respectively. Combing the results of the three analyses, we hypothesize that the C17 diynes inhibit fatty acid biosynthesis, specifically phospholipid synthesis in mycobacteira. Mycobacterium smegmatis MC2 155 was treated with 10 times of MIC90 of falcarinol, panaxydol, isoniazid, ethambutol and kanamycin for 6 hours with at least 6 independent biological replicates.
Project description:To elucidate the antivirulent lactone U1 mode of action, next generation sequencing was applied to analyze the transcriptome of NCTC 8325 cells treated with either compound or DMSO as control. Overall design: Two biological replicates for compound treated and also two biological replicates for DMSO treated Staphylococcus aureus cultures were analysed.
Project description:To elucidate the antivirulent hydroxy amide (R*,R*)-3 mode of action, next generation sequencing was applied to analyze the transcriptome of NCTC 8325 cells treated with either compound or DMSO as control. Overall design: Two biological replicates for compound treated and also two biological replicates for DMSO treated Staphylococcus aureus cultures were analysed.
Project description:Oxamniquine resistance evolved in the human blood fluke (Schistosoma mansoni) in Brazil in the 1970s. We crossed parental parasites differing ~500-fold in drug response, determined drug sensitivity and marker segregation in clonally-derived F2s, and identified a single QTL (LOD=31) on chromosome 6. A sulfotransferase was identified as the causative gene using RNAi knockdown and biochemical complementation assays and we subsequently demonstrated independent origins of loss-of-function mutations in field-derived and laboratory-selected resistant parasites. These results demonstrate the utility of linkage mapping in a human helminth parasite, while crystallographic analyses of protein-drug interactions illuminate the mode of drug action and provide a framework for rational design of oxamniquine derivatives that kill both S. mansoni and S. haematobium, the two species responsible for >99% of schistosomiasis cases worldwide. mRNA profiles from adult worms resistant (HR, 3 replicates) and susceptible (LE, 2 replicates) to the oxamniquine drug were compared to identify differential expression in genes related to drug resistance
Project description:Alpha-mangostin (α-MG) is a natural xanthone reported to exhibit rapid bactericidal activity against Gram-positive bacteria, and may therefore have potential clinical application in healthcare sectors. This study sought to identify the antibacterial mode of action of α-MG against Staphylococcus epidermidis RP62A through RNA-sequencing technology. Overall design: Overnight cultures of S. epidermidis were diluted 1:100 in LB and were grown to mid-log at 37 °C. Bacterial cells were treated with 0.7 MIC (0.875 µg/ml) of α-MG or 0.1 % methanol (solvent control) for 10 min. After incubation, bacterial total RNA was isolated. Samples were prepared in triplicates. The ribosomal RNA was depleted using Ribo-Zero rRNA Removal kit for Gram-positive bacteria. Sequencing library was prepared using the drop-seq protocol (Macosko et al., 2015) with specially designed random hexamer-based capture oligos with known barcodes. Paired-end sequencing was performed on NextSeq500 sequencer (Illumina) in FuGU (Helsinki, Finland).
Project description:Due to the rise of drug resistant forms of tuberculosis there is an urgent need for novel antibiotics to effectively combat these cases and to shorten treatment regimens. Recently, drug screens using whole cell analyses have shown to be successful. However, current high throughput screens focus mostly on stricto sensu life-death screening that give little qualitative information and often require the lengthy process of target and mode of action (MoA) identification. In doing so, promising compound scaffolds or non-optimized compounds that fail to reach inhibitory concentrations are missed. To accelerate early TB drug discovery, we performed RNA sequencing on Mycobacterium tuberculosis and Mycobacterium marinum to map the stress responses that follow upon exposure to sub-inhibitory concentrations of antibiotics with known targets: ciprofloxacin, ethambutol, isoniazid, streptomycin and rifampicin. The resulting dataset comprises the first overview of transcriptional stress responses of mycobacteria to different antibiotics. We show that antibiotics can be distinguished based on their specific transcriptional stress fingerprint i.e. DNA damage for ciprofloxacin and ribosomal stress for streptomycin. Notably, this fingerprint was more distinctive in M. marinum and we decided to use this to our advantage and continue with this model organism. A selection of diverse antibiotic stress genes was used to construct stress reporters. In total, three functional reporters were constructed for DNA damage, cell wall damage and ribosomal inhibition. Subsequently, these reporter strains were used to screen a small anti-TB compound library to predict the mode of action. In doing so we could identify the putative mode of action for three novel compounds, which confirms our approach. Overall design: RNA expression profiles of Mycobacterium tuberculosis and Mycobacterium marinum was generated upon exposure to sub-inhibitory concentrations of antibiotics with known targets: ciprofloxacin, ethambutol, isoniazid, streptomycin and rifampicin for 4 hrs and 24hrs duration and compared with controls without any drug exposure