Dataset Information

Transcriptome analysis reveals molecular anthelmintic effects of procyanidins in C. elegans

ABSTRACT: Worldwide, more than 1 billion people are affected by infestations with soil-transmitted helminths and also in veterinary medicine helminthiases are a severe thread to livestock due to emerging resistances against the common anthelmintics. Proanthocyanidins have been increasingly investigated for their anthelmintic properties, however, except for an interaction with certain proteins of the nematodes, not much is known about their mode of action. To investigate the anthelmintic activity on a molecular level, a transcriptome analysis was performed in Caenorhabditis elegans after treatment with purified and fully characterized oligomeric procyanidins (OPC). The OPCs had previously been obtained from a hydro-ethanolic (1:1) extract from the leaves of Combretum mucronatum, a plant which is traditionally used in West Africa for the treatment of helminthiasis, therefore, also the crude extract was included in the study. Significant changes in differential gene expression were observed mainly for proteins related to the intestine, many of which were located extracellularly or within cellular membranes. Among the up-regulated genes, several hitherto undescribed orthologues of structural proteins in humans were identified, but also genes that are potentially involved in the worms’ defense against tannins. For example, T22D1.2, an orthologue of human basic salivary proline-rich protein (PRB) 2, and numr-1 (nuclear localized metal responsive) were found to be strongly up-regulated. Down-regulated genes were mainly associated with lysosomal activity, glycoside hydrolysis or the worms’ innate immune response. No major differences were found between the groups treated with purified OPCs versus the crude extract. Investigations using GFP reporter gene constructs of T22D1.2 and numr-1 corroborated the intestine as the predominant site of the anthelmintic activity.

ORGANISM(S): Caenorhabditis elegans

PROVIDER: GSE101680 | GEO | 2017/08/31

SECONDARY ACCESSION(S): PRJNA395200

REPOSITORIES: GEO

ACCESS DATA

Dataset's files

Source:

			Action	DRS
		Other

Items per page:

1 - 1 of 1

Similar Datasets

Project description:Ascarid parasites infect a variety of hosts, and control measures rely heavily on anthelmintics raising concerns for anthelmintic resistance. Parascaris spp., the equine ascarid, is the only ascarid with documented substantial drug resistance. Elucidating drug responses of susceptible parasites may lead to further understanding of anthelmintic resistance mechanisms, identifying alternative drug targets, and parameters for slowing the development of resistance among other ascarid species. The purpose of this study was to examine the response of anthelmintic naïve Parascaris spp. to in vitro drug exposure using transcriptomic analyses and quantitative PCR. Adult Parascaris spp. worms were obtained at necropsy from foals born into a herd kept without anthelmintic treatment. Adult worms were maintained in Roswell-Park Memorial medium 1640. The drugs employed were oxibendazole (OBZ) and ivermectin (IVM), and 10-fold serial dilutions were prepared. Optimum drug concentration and exposure length were determined using a worm viability assessment method. Subsequently, RNA-seq analysis identified genes with significantly different expression between drug treated and control groups. Finally, a selection of the significant genes was further investigated using qPCR in a repeat drug exposure trial. Following initial viability assessments, OBZ (10 μg/mL) for 24 hours and IVM (1 μg/mL) for three hours were the conditions employed. RNA-seq analysis revealed a total of 88 transcripts with significantly (α=0.01) different expression between all drug treated and control worms. The top five genes selected for further analysis were snap25, cytochrome p450, kelch10, sup9, and ferm. Gene functionalities are related to muscle contraction, detoxification and exocytotic events, and regulatory processes. qPCR did not identify any significant differences between groups, but this may be due to the lack of method sensitivity. The results of RNA-seq analysis and gene function warrant further investigation and their role in ascarid xenobiotic responses.

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.

Dataset Information

Transcriptome analysis reveals molecular anthelmintic effects of procyanidins in C. elegans

Dataset's files

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets