Dataset Information

Veterinary trypanocidal benzoxaboroles are peptidase-activated prodrugs (T. congolense assay)

ABSTRACT:

Livestock diseases caused by Trypanosoma congolense, T. vivax and T. brucei, collectively known as nagana, are responsible for billions of dollars in lost food production annually. There is an urgent need for novel therapeutics. Encouragingly, promising antitrypanosomal benzoxaboroles are under veterinary development. Here, we show that the most efficacious subclass of these compounds are prodrugs activated by trypanosome serine carboxypeptidases (CBPs). Drug-resistance to a development candidate, AN11736, emerged readily in T. brucei, due to partial deletion within the locus containing three tandem copies of the CBP genes. T. congolense parasites, which possess a larger array of related CBPs, also developed resistance to AN11736 through deletion within the locus. A genome-scale screen in T. brucei confirmed CBP loss-of-function as the primary mechanism of resistance and CRISPR-Cas9 editing proved that partial deletion within the locus was sufficient to confer resistance. CBP re-expression in either T. brucei or T. congolense AN11736-resistant lines restored drug-susceptibility. CBPs act by cleaving the benzoxaborole AN11736 to a carboxylic acid derivative, revealing a prodrug activation mechanism. Loss of CBP activity results in massive reduction in net uptake of AN11736, indicating that entry is facilitated by the concentration gradient created by prodrug metabolism.

T. congolense assay is reported in the current study MTBLS1309.

T. brucei assay is reported in MTBLS1474.

Linked cross omic data sets:

Genome data associated with this study are available in the European Nucleotide Archive (ENA): accession number PRJEB34627.

INSTRUMENT(S): Orbitrap Fusion (Thermo Fisher Scientific)_negative, Orbitrap Fusion (Thermo Fisher Scientific)

SUBMITTER: Daniel Paape

PROVIDER: MTBLS1309 | MetaboLights | 2020-10-22

REPOSITORIES: MetaboLights

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Dataset's files

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			Action	DRS
	MTBLS1309	Other
	FILES	Other
	a_MTBLS1309_NEG_metabolite_profiling_mass_spectrometry.txt	Txt
	a_MTBLS1309_POS_metabolite_profiling_mass_spectrometry.txt	Txt
	i_Investigation.txt	Txt

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Publications

Veterinary trypanocidal benzoxaboroles are peptidase-activated prodrugs.

Giordani Federica F Paape Daniel D Vincent Isabel M IM Pountain Andrew W AW Fernández-Cortés Fernando F Rico Eva E Zhang Ning N Morrison Liam J LJ Freund Yvonne Y Witty Michael J MJ Peter Rosemary R Edwards Darren Y DY Wilkes Jonathan M JM van der Hooft Justin J J JJJ Regnault Clément C Read Kevin D KD Horn David D Field Mark C MC Barrett Michael P MP

PLoS pathogens 20201103 11

PMID: 33141865

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Project description:Livestock diseases caused by Trypanosoma congolense, T. vivax and T. brucei, collectively known as nagana, are responsible for billions of dollars in lost food production annually. There is an urgent need for novel therapeutics. Encouragingly, promising antitrypanosomal benzoxaboroles are under veterinary development. Here, we show that the most efficacious subclass of these compounds are prodrugs activated by trypanosome serine carboxypeptidases (CBPs). Drug-resistance to a development candidate, AN11736, emerged readily in T. brucei, due to partial deletion within the locus containing three tandem copies of the CBP genes. T. congolense parasites, which possess a larger array of related CBPs, also developed resistance to AN11736 through deletion within the locus. A genome-scale screen in T. brucei confirmed CBP loss-of-function as the primary mechanism of resistance and CRISPR-Cas9 editing proved that partial deletion within the locus was sufficient to confer resistance. CBP re-expression in either T. brucei or T. congolense AN11736-resistant lines restored drug-susceptibility. CBPs act by cleaving the benzoxaborole AN11736 to a carboxylic acid derivative, revealing a prodrug activation mechanism. Loss of CBP activity results in massive reduction in net uptake of AN11736, indicating that entry is facilitated by the concentration gradient created by prodrug metabolism. T. brucei assay is reported in the current study <a href='https://www.ebi.ac.uk/metabolights/MTBLS1474' rel='noopener noreferrer' target='_blank'>MTBLS1474</a>.T. congolense assay is reported in <a href='https://www.ebi.ac.uk/metabolights/MTBLS1309' rel='noopener noreferrer' target='_blank'>MTBLS1309</a>. Linked cross omic data sets:Genome data associated with this study are available in the European Nucleotide Archive (ENA): accession number <a href='https://www.ebi.ac.uk/ena/browser/view/PRJEB34627' rel='noopener noreferrer' target='_blank'>PRJEB34627</a>.

Project description:Testing the effect of seaweed as a novel cost-efficient feed source for Atlantic salmon feed. Fermentation of novel raw materials for fish feed is a new practice with little prior experience of direct relevance to aquaculture and fish farming. In Trial D, an alternative marine compound is included in the feed, which is based on seaweed. HoloFood Trial D - fermented seaweed open water-dose response is reported in the current study <a href='https://www.ebi.ac.uk/metabolights/MTBLS6733' rel='noopener noreferrer' target='_blank'>MTBLS6733</a>.HoloFood Trial A - seaweed-dose response is reported in <a href='https://www.ebi.ac.uk/metabolights/MTBLS4381' rel='noopener noreferrer' target='_blank'>MTBLS4381</a>.HoloFood Trial B - blue mussel-dose response is reported in <a href='https://www.ebi.ac.uk/metabolights/MTBLS4382' rel='noopener noreferrer' target='_blank'>MTBLS4382</a>.HoloFood Trial C - blue mussel ensilage-dose response is reported in <a href='https://www.ebi.ac.uk/metabolights/MTBLS4384' rel='noopener noreferrer' target='_blank'>MTBLS4384</a>.HoloFood Trial 1/2/3 is reported in <a href='https://www.ebi.ac.uk/metabolights/MTBLS6988' rel='noopener noreferrer' target='_blank'>MTBLS6988</a>. Linked cross omic data sets:Nucleic acid data associated with this study are available in the European Nucleotide Archive (ENA): accession number <a href='https://www.ebi.ac.uk/ena/browser/view/PRJEB43192' rel='noopener noreferrer' target='_blank'>PRJEB43192</a>.Metagenomic data associated with this study are available from MGnify under the Super Study '<a href='https://www.ebi.ac.uk/metagenomics/super-studies/holofood' rel='noopener noreferrer' target='_blank'>holofood</a>'.

Project description:Testing the effect of blue mussel ensilage as a novel cost-efficient protein feed source for Atlantic salmon feed. Fermentation of novel raw materials for fish feed is a new practice with little prior experience of direct relevance to aquaculture and fish farming. In Trial A, an alternative marine compound is included in the feed, which is based on seaweed. In the trial were multiple inclusion levels used to investigate the effect of seaweed inclusion in an aquaculture setting. HoloFood Trial A - seaweed-dose response is reported in the current study <a href='https://www.ebi.ac.uk/metabolights/MTBLS4381' rel='noopener noreferrer' target='_blank'>MTBLS4381</a>.HoloFood Trial B - blue mussel-dose response is reported in <a href='https://www.ebi.ac.uk/metabolights/MTBLS4382' rel='noopener noreferrer' target='_blank'>MTBLS4382</a>.HoloFood Trial C - blue mussel ensilage-dose response is reported in <a href='https://www.ebi.ac.uk/metabolights/MTBLS4384' rel='noopener noreferrer' target='_blank'>MTBLS4384</a>.HoloFood Trial D - fermented seaweed open water-dose response is reported in <a href='https://www.ebi.ac.uk/metabolights/MTBLS6733' rel='noopener noreferrer' target='_blank'>MTBLS6733</a>.HoloFood Trial 1/2/3 is reported in <a href='https://www.ebi.ac.uk/metabolights/MTBLS6988' rel='noopener noreferrer' target='_blank'>MTBLS6988</a>. Linked cross omic data sets:Nucleic acid data associated with this study are available in the European Nucleotide Archive (ENA): accession number <a href='https://www.ebi.ac.uk/ena/browser/view/PRJEB43192' rel='noopener noreferrer' target='_blank'>PRJEB43192</a>.Metagenomic data associated with this study are available from MGnify under the Super Study '<a href='https://www.ebi.ac.uk/metagenomics/super-studies/holofood' rel='noopener noreferrer' target='_blank'>holofood</a>'.

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Project description:Testing the effect of blue mussel ensilage as a novel cost-efficient protein feed source for Atlantic salmon feed. Fermentation of novel raw materials for fish feed is a new practice with little prior experience of direct relevance to aquaculture and fish farming. In Trial C, alternative processing of the raw blue mussel meat was tested where the protein was ensilaged before use instead of processed into a dry meal. By using this preservation process, a significant part of small peptides and free amino acids is generated. This trial aimed to examine whether the provision of better protein utilisation and a different microbiome in the gut of the fish can lead to a satisfactory utilisation of this ensilaged protein source by salmon. Earlier published results have shown that ensilaged fish protein provides better protein utilization and growth in Atlantic salmon. It is our assumption that the nutrient value of this product, therefore, is different compared to the more traditional protein meal tested in Trial B. It may also influence the microbiota in the gut differently based on the same arguments. The results from trial C will be very important as this ensilage-based treatment has not yet been used at a routine level, so any new information on its effect on fish health and growth will be highly useful to the aquaculture industry at large. HoloFood Trial C - blue mussel ensilage-dose response is reported in the current study <a href='https://www.ebi.ac.uk/metabolights/MTBLS4384' rel='noopener noreferrer' target='_blank'>MTBLS4384</a>.HoloFood Trial A - seaweed-dose response is reported in <a href='https://www.ebi.ac.uk/metabolights/MTBLS4381' rel='noopener noreferrer' target='_blank'>MTBLS4381</a>.HoloFood Trial B - blue mussel-dose response is reported in <a href='https://www.ebi.ac.uk/metabolights/MTBLS4382' rel='noopener noreferrer' target='_blank'>MTBLS4382</a>.HoloFood Trial D - fermented seaweed open water-dose response is reported in <a href='https://www.ebi.ac.uk/metabolights/MTBLS6733' rel='noopener noreferrer' target='_blank'>MTBLS6733</a>.HoloFood Trial 1/2/3 is reported in <a href='https://www.ebi.ac.uk/metabolights/MTBLS6988' rel='noopener noreferrer' target='_blank'>MTBLS6988</a>. Linked cross omic data sets:Nucleic acid data associated with this study are available in the European Nucleotide Archive (ENA): accession number <a href='https://www.ebi.ac.uk/ena/browser/view/PRJEB43192' rel='noopener noreferrer' target='_blank'>PRJEB43192</a>.Metagenomic data associated with this study are available from MGnify under the Super Study '<a href='https://www.ebi.ac.uk/metagenomics/super-studies/holofood' rel='noopener noreferrer' target='_blank'>holofood</a>'.

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