title: An in vivo zebrafish screen identifies organophosphate antidotes with diverse mechanisms of action.
abstract: Organophosphates are a class of highly toxic chemicals that includes many pesticides and chemical weapons. Exposure to organophosphates, either through accidents or acts of terrorism, poses a significant risk to human health and safety. Existing antidotes, in use for over 50 years, have modest efficacy and undesirable toxicities. Therefore, discovering new organophosphate antidotes is a high priority. Early life stage zebrafish exposed to organophosphates exhibit several phenotypes that parallel the human response to organophosphates, including behavioral deficits, paralysis, and eventual death. Here, we have developed a high-throughput zebrafish screen in a 96-well plate format to find new antidotes that counteract organophosphate-induced lethality. In a pilot screen of 1200 known drugs, we identified 16 compounds that suppress organophosphate toxicity in zebrafish. Several in vitro assays coupled with liquid chromatography/tandem mass spectrometry-based metabolite profiling enabled determination of mechanisms of action for several of the antidotes, including reversible acetylcholinesterase inhibition, cholinergic receptor antagonism, and inhibition of bioactivation. Therefore, the in vivo screen is capable of discovering organophosphate antidotes that intervene in distinct pathways. These findings suggest that zebrafish screens might be a broadly applicable approach for discovering compounds that counteract the toxic effects of accidental or malicious poisonous exposures.
authors.affiliation: Massachusetts General Hospital, Harvard Medical School, Department of Medicine, Charlestown, MA, USA.
authors.forename: Shan
authors.initials: S
authors.lastname: Jin
authors.forename: Kumar S
authors.initials: KS
authors.lastname: Sarkar
authors.forename: Youngnam N
authors.initials: YN
authors.lastname: Jin
authors.forename: Yan
authors.initials: Y
authors.lastname: Liu
authors.forename: David
authors.initials: D
authors.lastname: Kokel
authors.forename: Tjakko J
authors.initials: TJ
authors.lastname: Van Ham
authors.forename: Lee D
authors.initials: LD
authors.lastname: Roberts
authors.forename: Robert E
authors.initials: RE
authors.lastname: Gerszten
authors.forename: Calum A
authors.initials: CA
authors.lastname: Macrae
authors.forename: Randall T
authors.initials: RT
authors.lastname: Peterson
mesh_terms: D000818:Animals; D000931:Antidotes / Q000494:pharmacology*; D001285:Atropine / Q000494:pharmacology; D045744:Cell Line, Tumor; D018680:Cholinergic Antagonists / Q000494:pharmacology; D002801:Cholinesterase Reactivators / Q000494:pharmacology; D004353:Drug Evaluation, Preclinical; D004640:Emetine / Q000494:pharmacology; D006024:Glycopyrrolate / Q000494:pharmacology; D057166:High-Throughput Screening Assays; D006801:Humans; D007928:Lethal Dose 50; D008787:Metoclopramide / Q000494:pharmacology; D009388:Neostigmine / Q000494:pharmacology; D010755:Organophosphates / Q000633:toxicity*; D010278:Parathion / Q000633:toxicity*; D010890:Pirenzepine / Q000031:analogs & derivatives / Q000494:pharmacology; D011220:Pralidoxime Compounds / Q000494:pharmacology; D015027:Zebrafish
chemical_list: D000931:Antidotes; D018680:Cholinergic Antagonists; D002801:Cholinesterase Reactivators; D010755:Organophosphates; D011220:Pralidoxime Compounds; C046338:telenzepine; D009388:Neostigmine; D010890:Pirenzepine; D010278:Parathion; D001285:Atropine; D008787:Metoclopramide; C028797:pralidoxime; D006024:Glycopyrrolate; D004640:Emetine