T. cruzi resistant lines with compounds targeting cytochrome b
Ontology highlight
ABSTRACT:
PROVIDER: PRJEB47248 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| ERR6589050_1.fastq.gz | Fastqsanger.gz | |||
| ERR6589050_2.fastq.gz | Fastqsanger.gz | |||
| ERR6589051_1.fastq.gz | Fastqsanger.gz | |||
| ERR6589051_2.fastq.gz | Fastqsanger.gz | |||
| ERR6589052_1.fastq.gz | Fastqsanger.gz |
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Project description:Whole genome sequencing of Trypanosoma cruzi parasites resistant to DDD1542111
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Project description:<p>Chagas’ disease, caused by infection with the protozoan parasite Trypanosoma cruzi, is a potentially fatal condition for which new treatments are urgently needed. Due to the lack of validated drug targets, phenotypic screening followed by target deconvolution is the dominant approach in Chagas’ disease drug discovery. However, as most phenotypic screening hits act through a small number of promiscuous targets, implementation of counter-screening methodology for these targets as early as possible in the workflow is essential to enable prioritisation of compounds with novel Modes of Action (MoA). Here, we demonstrate that untargeted metabolomic profiling using liquid chromatography mass spectrometry (LC-MS) can reliably identify compounds that act through one of the most common targets, cytochrome b. Treatment of epimastigote form T. cruzi in culture with cytochrome b inhibitors resulted in rapid and pronounced perturbation of the metabolome. We identified a signature of 79 metabolites that were differentially expressed by at least 2-fold (p < 0.05). Unsupervised multivariate analysis using these features allowed clear separation of cytochrome b inhibitors from compounds acting through other MoA, and through disruption of oxidative phosphorylation by other mechanisms. Flexibility was observed in this cytochrome b signature between experiments, and depending on the compounds used, suggesting that this approach could be readily implemented in other laboratories. Triage of cytochrome b inhibitors early in the Chagas’ disease drug discovery workflow using untargeted metabolomics will aid in prioritisation of medicinal chemistry resources towards compounds acting through novel mechanisms.</p>
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Project description:Comparative genomic analysis of Trypanosoma cruzi G with Trypanosoma cruzi D11.
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Project description:As Trypanosoma cruzi, the etiological agent of Chagas disease, multiplies in the cytoplasm of nucleated host cells, infection with this parasite is highly likely to affect host cells. We performed an exhaustive transcriptome analysis of T. cruzi-infected HeLa cells using an oligonucleotide microarray containing probes for greater than 47,000 human gene transcripts. In comparison with uninfected cells, those infected with T. cruzi showed greater than threefold up-regulation of 41 genes and greater than threefold down-regulation of 23 genes. Real-time reverse transcriptase-polymerase chain reaction (RT-PCR) of selected, differentially expressed genes confirmed the microarray data. Many of these up- and down-regulated genes were related to cellular proliferation, including seven up-regulated genes encoding proliferation inhibitors and three down-regulated genes encoding proliferation promoters, strongly suggesting that T. cruzi infection inhibits host cell proliferation, which may allow more time for T. cruzi to replicate and produce its intracellular nests. These findings provide new insight into the molecular mechanisms by which intracellular T. cruzi infection influences the host cell, leading to pathogenicity. Keywords: infection response
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Project description:To generate a high quality, annotated gene expression database of T. cruzi trypomastigotes(TRP), amastigotes (AMA), epimastigotes (EPI), and metacyclic trypomastigotes (MET). The global assessment of transcript abundances in each life-cycle stage of T. cruzi will identify stage-regulated genes and provide an essential element of an integrated database of T. cruzi genomic, proteomic, and transcriptomic data. RNAs from 3 biological replicates from a single time point in each stage will be subjected to DNA microarrays containing probes for the complete, annotated T. cruzi genome, as it is currently known. The arrays for this project will be provided by the Pathogen Functional Genomics Resource Center (PFGRC) at The Institute for Genomic Research (TIGR) sponsored by the National Institute of Allergy and Infectious Diseases (NIAID), and will contain long oligonucleotides complementary to every annotated gene in the newly sequenced T. cruzi genome. The resulting data and analysis results will be deposited in TcruziDB (http://TcruziDB.org). Keywords: Gene expression comparisons between the four life-cycle stages of T. cruzi
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Project description:Proteomic study of SOAT1 protein targeting compounds in the treatment of liver cancer
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