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Caenorhabditis elegans is a useful model for anthelmintic discovery.

Burns AR, Luciani GM, Musso G, Bagg R, Yeo M, Zhang Y, Rajendran L, Glavin J, Hunter R, Redman E, Stasiuk S, Schertzberg M, Angus McQuibban G, Caffrey CR, Cutler SR, Tyers M, Giaever G, Nislow C, Fraser AG, MacRae CA, Gilleard J, Roy PJ - Nat Commun (2015)

Bottom Line: We then rescreen our hits in two parasitic nematode species and two vertebrate models (HEK293 cells and zebrafish), and identify 30 structurally distinct anthelmintic lead molecules.We identify the target of one lead with nematode specificity and nanomolar potency as complex II of the electron transport chain.This work establishes C. elegans as an effective and cost-efficient model system for anthelmintic discovery.

View Article: PubMed Central - PubMed

Affiliation: The Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Toronto, Ontario, Canada M5S 3E1.

ABSTRACT
Parasitic nematodes infect one quarter of the world's population and impact all humans through widespread infection of crops and livestock. Resistance to current anthelmintics has prompted the search for new drugs. Traditional screens that rely on parasitic worms are costly and labour intensive and target-based approaches have failed to yield novel anthelmintics. Here, we present our screen of 67,012 compounds to identify those that kill the non-parasitic nematode Caenorhabditis elegans. We then rescreen our hits in two parasitic nematode species and two vertebrate models (HEK293 cells and zebrafish), and identify 30 structurally distinct anthelmintic lead molecules. Genetic screens of 19 million C. elegans mutants reveal those nematicides for which the generation of resistance is and is not likely. We identify the target of one lead with nematode specificity and nanomolar potency as complex II of the electron transport chain. This work establishes C. elegans as an effective and cost-efficient model system for anthelmintic discovery.

No MeSH data available.


Related in: MedlinePlus

Wact-11 and wact-12 resistant mutants are cross-resistant to all nine wact-11-family members.(a) The wact-11-family core structure and the structure of an unrelated molecule, wact-2, which was used as a negative control throughout this work. (b) Heat maps of the wact-11-family dose-response experiments with wild-type worms (N2 strain), as well as two mutant strains, RP2674 and RP2698, isolated as being resistant to wact-12 and wact-11, respectively. The dose-response experiments were carried out using a 96-well plate liquid-based assay (see Methods). White indicates that there were more than 50 worms in three out of four replicate wells. Pink indicates that there were between 12 and 50 worms in three out of four replicate wells. Red indicates that there were between 0 and 11 worms in three out of four replicate wells. In the case of ties, the higher number prevailed (for example, at a given concentration, if two wells had 55 worms, and the other two wells had 20 worms, the chemical would be scored as having more than 50 worms). The R1 and R2 groups are indicated for each wact-11-family member. Wact-2 is used here as a negative control.
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f3: Wact-11 and wact-12 resistant mutants are cross-resistant to all nine wact-11-family members.(a) The wact-11-family core structure and the structure of an unrelated molecule, wact-2, which was used as a negative control throughout this work. (b) Heat maps of the wact-11-family dose-response experiments with wild-type worms (N2 strain), as well as two mutant strains, RP2674 and RP2698, isolated as being resistant to wact-12 and wact-11, respectively. The dose-response experiments were carried out using a 96-well plate liquid-based assay (see Methods). White indicates that there were more than 50 worms in three out of four replicate wells. Pink indicates that there were between 12 and 50 worms in three out of four replicate wells. Red indicates that there were between 0 and 11 worms in three out of four replicate wells. In the case of ties, the higher number prevailed (for example, at a given concentration, if two wells had 55 worms, and the other two wells had 20 worms, the chemical would be scored as having more than 50 worms). The R1 and R2 groups are indicated for each wact-11-family member. Wact-2 is used here as a negative control.

Mentions: Wact-11, wact-12 and wact-127 are three of the molecules against which we could generate resistant mutants (Supplementary Table 1). These three molecules are part of the C10 cluster (Fig. 2b), which we refer to as the ‘wact-11 family', and share an ethyl benzamide moiety (Fig. 3). In total, we isolated 37 mutants that resist the wact-11 family members at a rate of one mutant per 100,000 genomes screened (Supplementary Table 1). Using a representative set of 21 mutants, we performed a detailed dose-response analysis of each mutant against wact-11 and the structurally unrelated nematicide wact-2 in liquid media (Supplementary Fig. 4). All of the tested mutants show at least some resistance to wact-11, but not to wact-2, indicating that they are specifically resistant to wact-11.


Caenorhabditis elegans is a useful model for anthelmintic discovery.

Burns AR, Luciani GM, Musso G, Bagg R, Yeo M, Zhang Y, Rajendran L, Glavin J, Hunter R, Redman E, Stasiuk S, Schertzberg M, Angus McQuibban G, Caffrey CR, Cutler SR, Tyers M, Giaever G, Nislow C, Fraser AG, MacRae CA, Gilleard J, Roy PJ - Nat Commun (2015)

Wact-11 and wact-12 resistant mutants are cross-resistant to all nine wact-11-family members.(a) The wact-11-family core structure and the structure of an unrelated molecule, wact-2, which was used as a negative control throughout this work. (b) Heat maps of the wact-11-family dose-response experiments with wild-type worms (N2 strain), as well as two mutant strains, RP2674 and RP2698, isolated as being resistant to wact-12 and wact-11, respectively. The dose-response experiments were carried out using a 96-well plate liquid-based assay (see Methods). White indicates that there were more than 50 worms in three out of four replicate wells. Pink indicates that there were between 12 and 50 worms in three out of four replicate wells. Red indicates that there were between 0 and 11 worms in three out of four replicate wells. In the case of ties, the higher number prevailed (for example, at a given concentration, if two wells had 55 worms, and the other two wells had 20 worms, the chemical would be scored as having more than 50 worms). The R1 and R2 groups are indicated for each wact-11-family member. Wact-2 is used here as a negative control.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4491176&req=5

f3: Wact-11 and wact-12 resistant mutants are cross-resistant to all nine wact-11-family members.(a) The wact-11-family core structure and the structure of an unrelated molecule, wact-2, which was used as a negative control throughout this work. (b) Heat maps of the wact-11-family dose-response experiments with wild-type worms (N2 strain), as well as two mutant strains, RP2674 and RP2698, isolated as being resistant to wact-12 and wact-11, respectively. The dose-response experiments were carried out using a 96-well plate liquid-based assay (see Methods). White indicates that there were more than 50 worms in three out of four replicate wells. Pink indicates that there were between 12 and 50 worms in three out of four replicate wells. Red indicates that there were between 0 and 11 worms in three out of four replicate wells. In the case of ties, the higher number prevailed (for example, at a given concentration, if two wells had 55 worms, and the other two wells had 20 worms, the chemical would be scored as having more than 50 worms). The R1 and R2 groups are indicated for each wact-11-family member. Wact-2 is used here as a negative control.
Mentions: Wact-11, wact-12 and wact-127 are three of the molecules against which we could generate resistant mutants (Supplementary Table 1). These three molecules are part of the C10 cluster (Fig. 2b), which we refer to as the ‘wact-11 family', and share an ethyl benzamide moiety (Fig. 3). In total, we isolated 37 mutants that resist the wact-11 family members at a rate of one mutant per 100,000 genomes screened (Supplementary Table 1). Using a representative set of 21 mutants, we performed a detailed dose-response analysis of each mutant against wact-11 and the structurally unrelated nematicide wact-2 in liquid media (Supplementary Fig. 4). All of the tested mutants show at least some resistance to wact-11, but not to wact-2, indicating that they are specifically resistant to wact-11.

Bottom Line: We then rescreen our hits in two parasitic nematode species and two vertebrate models (HEK293 cells and zebrafish), and identify 30 structurally distinct anthelmintic lead molecules.We identify the target of one lead with nematode specificity and nanomolar potency as complex II of the electron transport chain.This work establishes C. elegans as an effective and cost-efficient model system for anthelmintic discovery.

View Article: PubMed Central - PubMed

Affiliation: The Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Toronto, Ontario, Canada M5S 3E1.

ABSTRACT
Parasitic nematodes infect one quarter of the world's population and impact all humans through widespread infection of crops and livestock. Resistance to current anthelmintics has prompted the search for new drugs. Traditional screens that rely on parasitic worms are costly and labour intensive and target-based approaches have failed to yield novel anthelmintics. Here, we present our screen of 67,012 compounds to identify those that kill the non-parasitic nematode Caenorhabditis elegans. We then rescreen our hits in two parasitic nematode species and two vertebrate models (HEK293 cells and zebrafish), and identify 30 structurally distinct anthelmintic lead molecules. Genetic screens of 19 million C. elegans mutants reveal those nematicides for which the generation of resistance is and is not likely. We identify the target of one lead with nematode specificity and nanomolar potency as complex II of the electron transport chain. This work establishes C. elegans as an effective and cost-efficient model system for anthelmintic discovery.

No MeSH data available.


Related in: MedlinePlus