Unknown,Transcriptomics,Genomics,Proteomics

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Expression data from Caenorhabditis elegans exposed to albendazole

ABSTRACT: We have investigated how the model nematode Caenorhabditis elegans responds to and metabolizes albendazole; an important anthelmintic for human and animal parasite control. The transcriptional response of the mutant strain CB3474 ben-1(e1880)III, which is highly resistant to benzimidazoles due to a null mutation in the Î²-tubulin drug target, was examined. This approach was successful in minimizing transcriptional responses associated with non-specific stress or with the drug mode of action, resulting in only in a small subset of genes showing differential expression in response to drug exposure. Matched cultures of synchronised C. elegans were grown to young adult stage in liquid culture. The nematodes were then exposed to 300 ug/ml albendazole (ABZ) or exposed only to the DMSO excipient used to deliver the albendazole (CONT) for 4 hours. RNA was extracted from three biological replicates and hybridised to Affymetrix arrays.

ORGANISM(S): Caenorhabditis elegans

SUBMITTER: Al Ivens

PROVIDER: E-GEOD-21747 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Infections and diseases caused by parasitic nematodes have a major adverse impact on the health and productivity of animals and humans worldwide. The control of these parasites often relies heavily on the treatment with commercially available chemical compounds (anthelmintics). However, the excessive or uncontrolled use of these compounds in livestock animals has led to major challenges linked to drug resistance in nematodes. Therefore, there is a need to develop new anthelmintics with novel mechanism(s) of action. Recently, we identified a small molecule, KM08948, with nematocidal activity against the free-living model organism Caenorhabditis elegans. Here, we evaluated KM08948’s potential as an anthelmintic in a structure-activity relationship (SAR) study in C. elegans and in the highly pathogenic, blood-feeding Haemonchus contortus (barber’s pole worm), and explored the compound-target relationship using thermal proteome profiling (TPP). First, we synthesised and tested a series of 25 KM08948 analogues for nematocidal activity in both H. contortus (larvae and adults) and C. elegans (young adults), establishing a preliminary nematocidal pharmacophore for both species. We identified several compounds with marked activity against either H. contortus or C. elegans which had greater efficacy than KM08948, and found a significant divergence in compound bioactivity between these two nematode species. We also identified a KM08948 analogue, 25, that moderately inhibited the motility of adult female H. contortus in vitro. Subsequently, we inferred three H. contortus proteins (HCON_00134350, HCON_00021470 and HCON_00099760) and five C. elegans proteins (F30A10.9, F15B9.8, B0361.6, DNC-4 and UNC-11) that interacted directly with KM08948; however, no conserved protein target was shared between the two nematode species. Future work aims to extend the SAR investigation in these and other parasitic nematode species, and validate individual proteins identified here as targets of KM08948. Overall, the present study describes an evaluation of this anthelmintic candidate and highlights some challenges associated with early anthelmintic discovery.

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Project description:Quantitative high-throughput screening (qHTS) pharmacologically evaluates libraries of drugs and investigational agents for potential therapeutic uses, toxicological risk assessment, and increasingly for academic chemical tool discovery. Phenotypic HTS assays aim to interrogate molecular pathways and networks, often relying on cell culture systems, historically with less emphasis on multicellular organisms. C. elegans has served as a powerful eukaryotic model organism for human biology and disease by virtue of genetic conservation and experimental tractability. Here we describe a paradigm to enable C. elegans in qHTS using 384-well microtiter plate laser scanning cytometry for rapid signal acquisition with concurrent quantification of the fluorescent protein-expressing organism to select phenotype-modifying compounds for subsequent life-stage and proteomic analysis. E. coli bacterial ghosts, a non-replicating nutrient, allow compound exposures over 7-days spanning two life cycles to mitigate complications from bacterial overgrowth. We demonstrate the method using 643 anti-infective biased agents tested in 7-pt titration to assess feasibility of nematode-based in vivo qHTS. A pharmacological profile from the primary screen confirmed the efficacy of known anti-parasitic molecules, such as ivermectin and levamisole as well as illuminating anthelmintic properties of general chemical classes, including MEK, -secretase, bromodomain and proteasome inhibitors. Subsequent life-stage and proteomics analysis helped elucidate the physiologic consequence of specific compounds on C. elegans viability. For example, bortezomib induced a parallel upregulation of core proteasome subunits and the transcription factor skn-1-activating DDI aspartyl protease, while the kinase inhibitor trametinib upregulated the C. elegans MEK-1 ortholog, Mnk-1, downstream aminoacyl tRNA synthetases and ribosomal survival pathway proteins. We anticipate a broader application of this qHTS-coupled proteomics approach will enable the analysis of C. elegans orthologous transgenic phenotypes of human pathologies to facilitate drug and probe profiling from high-impact chemical libraries for a range of therapeutic indications.

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Expression data from Caenorhabditis elegans exposed to albendazole

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