Novel behavioral screening tool for therapeutics against organophosphorus agents

用于有机磷药物治疗的新型行为筛选工具

• 批准号: 10631009
• 负责人: Eva-Maria Schoetz Collins
• 金额: $ 19.23万
• 依托单位: SWARTHMORE COLLEGE
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10631009
• 关键词: Acceleration; Accidents; Acetylcholine; Acetylcholinesterase; Acute; Adopted; Adult; Adverse effects; Aftercare; Age; Animals; Anticonvulsants; Atropine; Behavior; Behavioral; Biological Assay; Brain Death; Brain Injuries; Butyrylcholinesterase; Carbamates; Characteristics; Chemical Exposure; Chemicals; Cholinesterases; Chronic; Comparative Study; Complex; Data; Development; Dose; Enzyme Inhibition; Enzymes; Epilepsy; Exposure to; Glutamate Agonist; Glutamate Receptor; Goals; Health; Hour; Human; In Vitro; Individual; Industrial Accidents; Invertebrates; Length; Life; Mammals; Measures; Medical; Metabolic; Methods; Midazolam; Morphology; Muscarinic Acetylcholine Receptor; N-Methylaspartate; Neurotoxins; Nicotinic Receptors; Organism; Organophosphorus Compounds; Oximes; Parathion; Pesticides; Pharmaceutical Preparations; Phenotype; Physostigmine; Planarians; Platyhelminths; Poisoning; Population; Proteins; Rapid screening; Research; Respiratory distress; Robotics; Screening procedure; Seizures; Status Epilepticus; Survival Rate; Symptoms; System; Terrorism; Testing; Therapeutic; Therapeutic Uses; Time; Toxic effect; Unconscious State; acute symptom; acute toxicity; cholinergic; cost; cost effective; effective therapy; fluorophosphate; high throughput screening; improved; knock-down; mass casualty; meter; mortality; nerve gas; neuronal circuitry; novel; novel therapeutics; pesticide exposure; pesticide poisoning; prevent; reduce symptoms; screening; therapeutic candidate; therapeutic evaluation; toxicant; treatment strategy

Objectives. Terrorist acts or accidental poisoning involving acute exposure to organophosphorus (OP) agents, such as nerve gases and OP pesticides (OPPs), pose a serious threat to induce mass casualties. Acute OP toxicity results from inhibition of acetylcholinesterase (AChE), causing acetylcholine accumulation and cholinergic crisis. Symptoms include increased secretions, respiratory distress, loss of consciousness, and status epilepticus, causing permanent brain damage and death if untreated. Acute OP toxicity is currently treated with atropine and pralidoxime (2-PAM) to mitigate cholinergic hyperstimulation, and midazolam to stop epileptic seizures. This treatment reduces mortality, but only if administered within minutes of exposure and is thus not realistic for treating the civilian population during a mass casualty. Moreover, adverse health effects may remain after treatment. Thus, there is an urgent need for inexpensive therapeutics that can reduce mortality and alleviate adverse effects when administered later. The rigor of the prior research is hampered because current mammalian screening methods are slow and expensive, restricting the number of candidates that can be tested. High- throughput screening (HTS) platforms that can rapidly and cheaply screen possible candidates promise to accelerate the development of new therapeutics. The overarching goal of this research is to develop a cost- effective HTS organismal platform to streamline and accelerate first-tier screening of possible therapeutics using the planarian D. japonica. The specific objective of this proposal is to test the hypothesis that planarian HTS is “fit for purpose” as a screening tool for therapeutic candidates that can alleviate the symptoms of acute OPP exposure. Planarians are small flatworms with tractable, evolutionarily conserved neuronal circuits and a wide repertoire of complex behaviors that are amenable to HTS. As invertebrates, they are considered a non-animal organism. Unique to the planarian system, developing organisms are metabolically competent and can be screened from exposure onset into adulthood, allowing for the assessment of toxicants in mixed populations. Experimental approach. In Aim 1 will study the acute toxicity of 2 OPPs of concern, diisopropyl fluorophosphate and parathion. Using HTS, we will characterize their toxicity profiles and evaluate lethality, morphology and behavior in adult planarians at 30 minutes and 24 hours of exposure. In aim 2, we will verify that OPP-induced seizures in planarians are true seizures by developing quantitative metrics for comparison with verified planarian seizures and blocking of OPP-induced seizures with midazolam. In aim 3, we will demonstrate that acute OPP toxicity in planarians is responsive to therapeutics and that OPP-induced symptoms can be alleviated using combinatory treatment with atropine, 2-PAM, and midazolam, to establish a baseline to compare to novel drugs. Expected results. This proposal will demonstrate the value of planarian HTS for testing therapeutic candidates to treat OPP acute toxicity. The development of this rapid organismal screening method could be easily adopted to other test paradigms, including different life stages, lengths of exposure, and chemical domains.

目标.涉及急性接触有机磷制剂的恐怖行为或意外中毒， 例如神经毒气和有机磷农药，构成了造成大规模伤亡的严重威胁。急性OP 毒性是由于乙酰胆碱酯酶（AChE）受到抑制，导致乙酰胆碱积累， 胆碱能危象症状包括分泌物增加，呼吸窘迫，意识丧失， 癫痫持续状态，如果不治疗会导致永久性脑损伤和死亡。目前治疗急性OP毒性 阿托品和解磷定（2-PAM）减轻胆碱能过度刺激，咪达唑仑停止癫痫发作 癫痫发作这种治疗降低了死亡率，但只有在暴露后几分钟内给药， 在大规模伤亡中治疗平民是现实的。此外，对健康的不利影响可能仍然存在， 治疗后因此，迫切需要可以降低死亡率并减轻癌症的廉价治疗剂。 不良反应，以后服用。先前研究的严谨性受到阻碍，因为目前的哺乳动物 筛选方法既缓慢又昂贵，限制了可供测试的候选人数量。高- 可以快速且廉价地筛选可能的候选物的通量筛选（HTS）平台承诺， 加速新疗法的开发。这项研究的首要目标是开发一种成本- 有效的HTS有机平台，以简化和加速可能的治疗方法的一线筛选， 涡虫D.日本。这项建议的具体目标是测试这一假设，即涡虫HTS是 “适合目的”作为筛选可缓解急性脑梗死症状的治疗候选药物的工具 exposure. Planarians是一种小型的扁形虫，具有易于驾驭的，进化上保守的神经回路和广泛的神经元。 复杂行为的集合，这些行为符合HTS。作为无脊椎动物，它们被认为是非动物 有机体独特的涡虫系统，发展有机体的代谢能力，可以 从接触开始到成年进行筛查，以便对混合人群中的有毒物质进行评估。 实验方法。在目标1中，将研究2种相关OPP的急性毒性， 和双氯离子。使用HTS，我们将描述其毒性特征，并评价致死率、形态学和 在30分钟和24小时的暴露下，成年真涡虫的行为。在目标2中，我们将验证OPP诱导的 通过开发定量指标与经验证的真涡虫进行比较， 癫痫发作和用咪达唑仑阻断OPP诱导的癫痫发作。在目标3中，我们将证明， 涡虫中的毒性对治疗剂有反应，并且OPP诱导的症状可以使用 使用阿托品、2-PAM和咪达唑仑的组合治疗，以建立与新药比较的基线。 预期成果。这项建议将证明的价值，涡虫高温超导测试治疗的候选人 治疗急性毒性。这种快速生物筛选方法的发展， 其他测试范例，包括不同的生命阶段，暴露时间和化学领域。