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Investigating Cell Type Specific Mechanisms Contributing to Acute Oral Toxicity

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Investigating Cell Type Specific Mechanisms Contributing to Acute Oral Toxicity Prieto et al.: Investigating cell type specific mechanisms contributing to acute oral toxicity Supplementary Data1 Tab. S1: Overview of collected information on specific target organ/system and general cytotoxicity for chemicals correctly assigned to the CLP acute oral toxicity category by the 3T3 NRU cytotoxicity assay General Nervous Cardiovascular GI Chemical Liver Lung Blood Kidney cytotoxicity system system system (±)-Propranolol hydrochloride x ax (4-Ammonio-m-tolyl)ethyl(2- x x hydroxyethyl)ammonium sulfate 1,2,4-Trichlorobenzene x x 1,2-Dichlorobenzene x x 2,4,6- x x Tris(dimethylaminomethyl)phenol 2,4-Dichlorophenoxyacetic acid x x x 5,5-Diphenylhydantoin x x 5-Fluorouracil x x x x Acetophenone x Acetylsalicylic acid x x x x x x Acrolein x x Acrylamide x x x Ammonium chloride x ax ax Atropine sulfate monohydrate x x Benzaldehyde x x Cadmium (III) chloride x x x x x Caffeine x x x Chloroform x x ax x x x x Chloroquine bis(phosphate) x x Chlorpromazine x x x Codeine x x x x Colchicine x x x x Copper sulfate x x x Cupric sulfate pentahydrate x x Cyclosporin A x x x x Diazepam x Diphenhydramine hydrochloride x x Disopyramide x x Ethyl chloroacetate x x Ferrous sulfate x x x Glufosinate-ammonium x Glutethimide x ax x Hexachlorophene x x Lithium carbonate x x x Lithium sulfate x x x Malathion x x Maleic acid x x Meprobamate x x Orphenadrine hydrochloride x x x x p-Benzoquinone x x x Phenol x x x x x Procainamide hydrochloride x x x Quinidine sulfate dehydrate x x x doi:10.14573/altex.1805181s2 ALTEX 36(1), SUPPLEMENTARY DATA 1 General Nervous Cardiovascular GI Chemical Liver Lung Blood Kidney cytotoxicity system system system Resorcinol x x Rifampicin x x Sodium Cyanate x x Sodium oxalate x x x Sodium valproate x bx x x Thioridazine hydrochloride x x Valproic acid x x x x x GI: Gastrointestinal; CLP: Classification, labelling and packaging; NRU: Neutral Red Uptake; a Indirect effect: b chronic effect Tab. S2: Overview of collected information on specific target organ/system and general cytotoxicity for chemicals with CLP acute oral toxicity category under-predicted by the 3T3 NRU cytotoxicity assay General Nervous Cardiovascular GI Immune Chemical Liver Lung Blood Kidney cytotoxicity system system system system Potassium cyanide x x x (±)-Verapamil x hydrochloride 1-Phenyl-2-thiourea x x 1-Phenyl-3- x x pyrazolidone Barium chloride x x ax x Brucine x x x Chloral hydrate x x x x x cis- Diammineplatinum x x (II) dichloride D-Amphetamine x Dichlorvos x x x Disulfoton x Endosulfan x x Epinephrine x x hydrogen tartrate Fenpropathrin x Lindane x x x x Malononitrile x x ax x x x x x Mercury II chloride x bx x Methadone x x hydrochloride Nicotine x x Ochratoxin A x x x Paraquat dichloride x x x x Parathion ax x Pentachlorophenol x x x x Phenobarbital ax x ax Physostigmine x Sodium pentobarbital ax x Sodium salt of x x x chloroacetic acid Strychnine x x Thallium sulfate x x x Theophylline x x x x Triethylenemelamine x x x Warfarin x x GI: Gastrointestinal; CLP: Classification, labelling and packaging; NRU: Neutral Red Uptake; a indirect effect; b corrosive 2 Tab. S3: Overview of collected information on specific target organ/system and general cytotoxicity for chemicals with CLP acute oral toxicity category over-predicted in the 3T3 NRU cytotoxicity assay Nervous Cardiovascular GI Immune Chemical Cytotoxicity Liver Lung Blood Kidney system system system system 17α- x x x Ethynyloestradiol 1-Naphthylamine x Amitriptilyne x x x hydrochloride Diquat dibromide x x x x Haloperidol x x Maprotiline x x N-isopropyl-N'- phenyl-p- x phenylenediamine Tert-butyl x x x hydroperoxide Triphenyltin x hydroxide GI: Gastrointestinal; CLP: Classification, labelling and packaging; NRU: Neutral Red Uptake Tab. S4: Specific target organ/systems and general cytotoxicity reported for the chemicals falsely predicted as negatives (i.e. LD50 > 2000 mg/kg) by the 3T3 NRU cytotoxicity assay General Nervous Cardiovascular GI Chemical Liver Lung Kidney cytotoxicity system system system Aconitine x x x Isoniazid x x x Diethylene x x x glycol Digoxin x Ethylene glycol x x x x Paraldehyde x x GI: Gastrointestinal; NRU: Neutral Red Uptake 3 Tab. S5: Adverse outcome pathways outlined or fully developed with potential relevance to acute oral toxicity Adverse outcome Molecular initiating event (MIE) / Stressor References Key events (KEs) Cell death • Decompartmentalization Vinken and (Apoptosis/necrosis) • Direct mitochondrial inhibition Blaauboer, 1(AOP 205) • Narcosis 2017 In support of cytotoxicity as • Mitochondrial impairment (key event) general mechanism (non-cell specific) Hematotoxicity • Parent compound is converted to the 1(AOP 31) reactive metabolite and forms free radicals leading to oxidation of heme iron(II) in hemoglobin to iron(III) • Altered regulation of alpha- hemoglobin • Oxidative stress propagation • Damaging of red blood cells; hemolysis • Formation of hemoglobin adducts • Down regulation of glucose-6- phosphate dehydrogenase • Increase RBC congestion in liver • Increase liver and splenic hemosiderosis • Decrease in hemoglobin, hematocrit, and red blood cell number (methemoglobinemia) Acute liver response • PXR/SXR activation 1(AOP 11) • Key events unknown Cholestatic liver injury • Inhibition of bile salt export pump Vinken et al., 1(AOP 27) (ABCB11) 2013 • Activation of specific nuclear receptors, transcriptional change • Bile accumulation • Cytokine release • Increased inflammation • Reactive oxygen species production • Peptide oxidation Liver inflammation • Lysosome disruption • Iron compounds 1(AOP 144) • Peptide oxidation • ROS • Mitochondrial dysfunction 1 • o-Methyl-serine • Cell injury/death dodecylamide • Cytokine release hydrochloride (MSDH) • Inflammatory cells • alpha-Tocopheryl succinate • 3-Aminopropanal • Artesunate • Naphtharazine • Fluoroquinolones Decrease lung function • EGFR activation • ROS 1(AOP 148) • Trans differentiation of ciliated epithelial cells • Metaplasia of goblet cells • Goblet cell hyperplasia • Proliferation of goblet cells • SP1 activation • Apoptosis of ciliated epithelial cells • Increased mucin production • Chronic mucus hypersecretion Increased mortality (Impaired • Impaired ion channels heart function) • Altered action potential 1(AOP 104) • Increased cardiac arrythmia Hypertension • Peptide oxidation • ROS 1(AOP 149) • S-Glutathionylation, eNOS • Decrease in GTPCH-1 • Decrease in tetrahydrobiopterin 4 Adverse outcome Molecular initiating event (MIE) / Stressor References Key events (KEs) • Uncoupling of eNOS • Depletion of nitric oxide • Impaired vasodilation • Increase in vascular resistance • Decrease in AKT/eNOS activity Learning and impaired • Binding of agonist to ionotropic • Domoic acid memory glutamate receptors 4(AOP 48) • Mitochondrial dysfunction 1 • Cell injury/death • Neurodegeneration • Overactivation of NMDARs • Intracellular calcium overload • Decrease in neuronal network function in adult brain • Neuroinflammation Epileptic seizures • Binding at picrotoxin site of iGABAR • Lindane 2(AOP 10) chloride channel • Endosulfan • Reduction in ionotropic GABA receptor • Picrotoxin chloride channel conductance • Dieldrin • Reduction of neuronal synaptic • Heptachlor inhibition • RDX • Generation of amplified excitatory • Fipronil postsynaptic potential (EPSP) • Occurrence of a paroxysmal depolarizing shift Parkinsonian motor deficits • Binding of inhibitor to NADH- • MPP+ 3(AOP 3) ubiquinone oxidoreductase (complex I) • Inhibition of NADH-ubiquinone oxidoreductase (complex I) • Mitochondrial dysfunction 1 • Impaired proteostasis • Neuroinflammation • Degeneration of dopaminergic neurons of the nigrostriatal pathway Acute mortality • Inhibition of acetylcholinesterase • Organophosphates Russom et 1(AOP 16) (AchE) • Carbamates al., 2014 • Accumulation of acetylcholine in synapses • Increased atrioventricular block and bradycardia • Increased respiratory distress/arrest • Induction of ataxia, paralysis, or hyperactivity Acute mortality • Modulation of sodium channel • Cypermethrin 1(AOP 96) • Prolonged depolarization of neuronal • Permethrin membrane • Esfenvalerate • Neurotransmitter release • Tralomethrin • Muscle contraction • Bifenthrin • Increased ataxia, paralysis, or • Cyfluthrin hyperactivity • lambda-Cyhalothrin Neurodegeneration • Binding to SH/seleno proteins • Mercuric chloride 1(AOP 17) • Oxidative stress • Acrylamide • Glutamate dyshomeostasis • Acrolein • Cell injury/death • Methylmercuric(II) • Neuroinflammation chloride • Tissue resident cell activation • Thiomersal • Increase of pro-inflammatory mediators • Decrease of neuronal network function The acute neurotoxicity • Binding to the alpha subunit of voltage- • Pyrethroid Bal-Price et induced by binding of gated sodium channels al., 2017 pyrethroid insecticides to • Changes in the kinetics of channel voltage-gated sodium opening and closing channels • Alterations in excitability of neuronal membranes 5 Adverse outcome Molecular initiating event (MIE) / Stressor References Key events (KEs) (not in AOP-Wiki) • Dysregulation of neural networks Delayed neuropathy • Binding to neuropathy target esterase • Organophosphates Bal-Price et (NTE) al., 2015 Colony loss/failure • MIE unknown 1(AOP 84) • Suppression of the immune system • Increased viral susceptibility • Impaired development Colony loss/failure • MIE unknown 1(AOP 85) • Suppression of the immune system • Increased viral susceptibility • Abnormal foraging activity and behavior Inhibition/activation of gastric • MIE unknown • Nonsteroidal anti- ulcer formation • Inhibition
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