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（OP）制剂的恐怖行为或意外中毒， 神经毒气和有机磷杀虫剂 (OPP) 等，对造成大规模急性有机磷中毒构成严重威胁。 毒性是由于抑制乙酰胆碱酯酶（AChE）引起的，导致乙酰胆碱积累和 症状包括分泌物增多、呼吸困难、意识丧失和 癫痫持续状态，如果不及时治疗，会导致永久性脑损伤和死亡。目前正在治疗急性 OP 毒性。 与阿托品和解磷定 (2-PAM) 一起使用以减轻胆碱能过度刺激，并与咪达唑仑一起使用来阻止癫痫 这种治疗可以降低死亡率，但前提是在暴露后几分钟内进行，因此不能。 在大规模伤亡期间治疗平民是现实的。此外，不利的健康影响可能仍然存在。 因此，迫切需要能够降低死亡率和缓解症状的廉价疗法。 由于目前的哺乳动物，先前研究的严谨性受到阻碍。 筛选方法缓慢且昂贵，限制了可测试的候选人数量。 吞吐量筛选（HTS）平台可以快速、廉价地筛选可能的候选者，有望 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 生物平台，可简化和加速对可能的治疗方法的一级筛选 该提案的具体目标是检验涡虫 HTS 的假设。 “适合目的”作为候选治疗筛选工具，可缓解急性 OPP 症状 涡虫是一种小型扁虫，具有易于驯服、进化上保守的神经回路和广泛的暴露。 作为无脊椎动物，它们被认为是非动物。 涡虫系统所独有的，发育中的生物体具有代谢能力，并且可以 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 目标 1 将研究 2 种受关注的 OPP（氟磷酸二异丙酯）的急性毒性。 使用 HTS，我们将表征它们的毒性特征并评估致死率、形态和毒性。 成年涡虫暴露 30 分钟和 24 小时后的行为 在目标 2 中，我们将验证 OPP 诱导的情况。 通过制定定量指标与经过验证的涡虫进行比较，涡虫的癫痫发作是真正的癫痫发作 癫痫发作和用咪达唑仑阻断 OPP 引起的癫痫发作 在目标 3 中，我们将证明急性 OPP。 涡虫的毒性对治疗有反应，OPP 引起的症状可以通过使用来缓解 阿托品、2-PAM 和咪达唑仑联合治疗，以建立与新药进行比较的基线。 预期结果 该提案将证明涡虫 HTS 对于测试候选治疗药物的价值。 这种快速生物筛选方法的开发可以很容易地采用。 其他测试范例，包括不同的生命阶段、暴露时间和化学领域。