Acetylcholinesterase Complex Protein-Protein Interactions as Drug Targets Against Organophosphate-induced Neurotoxicity.

乙酰胆碱酯酶复合物蛋白质-蛋白质相互作用作为抗有机磷诱导的神经毒性的药物靶点。

• 批准号: 10772738
• 负责人: Jeremy Wayne Chambers
• 金额: $ 1.62万
• 依托单位: FLORIDA INTERNATIONAL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-05-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10772738
• 关键词: Acetylcholinesterase; Acetylcholinesterase Inhibitors; Acute; Adaptor Signaling Protein; Administrative Supplement; Age; Cause of Death; Cessation of life; Chemical Weapons; Chronic; Complex; Drug Targeting; Elements; Environmental Health; Enzymes; Exposure to; Goals; Impairment; In Vitro; Insecticides; Internships; Muscle; Muscle Contraction; Muscular Atrophy; Nervous System Trauma; Organophosphates; Oximes; Parathion; Pesticides; Phosphorylation; Protein Kinase; Protein Kinase Interaction; Proteins; Publishing; Recovery; Refractory; Research; Resistance; Retro-Inverso Peptide; Sarin; Seizures; Site; Small RNA; Students; Surface; Synapses; Therapeutic; Toxic effect; United States; aged; blood-brain barrier permeabilization; cholinergic; drug discovery; health science research; high school; in vivo; innovation; insight; nerve agent; neuromuscular; neurotoxicity; neurotransmission; novel strategies; parent grant; peptidomimetics; postsynaptic; protein complex; protein protein interaction; summer internship; toxic organophosphate insecticide exposure

Project Summary. This administrative supplement will introduce environmental health sciences research to a summer high school intern. Organophosphate (OP) insecticides (e.g., Naled, parathion) and chemical weapons (e.g., sarin, VX, Novichok) are robust inhibitors of acetylcholinesterase (AChE) triggering cholinergic crises resulting in muscle contractions, seizures, and in extreme exposures death. Certain OPs can rapidly (within minutes) irreversibly inhibit AChE, producing an “aged” species refractory to oxime reactivators, like 2-pralidoxime (2-PAM), which is part of the OP treatment in the United States and whose efficacy has recently been questioned. Consequently, there is an urgent need to discover novel approaches to rescue AChE activity following OP exposure to mitigate the damaging neurological and muscular effects. The long-term goal of the proposed research is to develop viable, lasting treatments for acute and chronic OP exposures. Previous efforts to rescue aged AChE have focused on producing new reactivators that will alkylate the phosphorylated enzyme allowing oximes like 2-PAM to recover enzyme function, but these compounds have shortcomings such as insufficient blood-brain barrier permeability and limited efficacy in vivo. The proposed research is innovative because it employs strategies aimed at increasing the turnover aged AChE and recovery of nascent intracellular AChE. For example, inhibiting or loss of muscle-specific protein kinase (MuSK) destabilizes the AChE complex leading to the degradation of the enzyme. Recent studies demonstrate that Dok7, an adaptor protein in the AChE complex, is necessary for MuSK activity and more importantly, AChE localization on the muscle surface. Therefore, the current research objective is to assess whether targeting proteins in the synaptic AChE complex can increase AChE turnover and restore optimal AChE activity. The central hypothesis in pursuit of this objective is that inhibiting or degrading proteins associated with aged AChE will cause the release of the aged enzyme, making way for new AChE to repopulate the synapse, restore optimal neurotransmission, and mitigate the effects of OP exposures. The rationale for the proposed research is that eliminating aged AChE will alleviate the detrimental effects of OP toxicity and restore proper neurotransmission. Based on published and preliminary studies, the high school intern will investigate elements in Specific Aim 1 of the parent grant: Evaluate the in vitro therapeutic benefit of targeting proteins in the AChE complex for degradation. In the proposed project, the high school intern will use small RNAs to diminish the protein levels of Dok7. Similarly, the student will use a retro-inverso peptide mimicking the Dok7- MuSK interaction site to destabilize the protein-protein interaction, which we hypothesize will result in the release of aged AChE These studies will determine the druggability of Dok7 and determine if it will be evaluated as drug discovery studies. This research is significant because it will provide fundamental mechanistic insights into the stability and turnover of post-synaptic AChE complexes and unearth new targets to treat OP exposures.

项目摘要。 这种行政补充剂将向夏季高中介绍环境健康科学研究 实习生。有机磷酸盐（OP）杀虫剂（例如，NaLed，Parathion）和化学武器（例如Sarin，VX，， Novichok）是乙酰胆碱酯酶（ACHE）触发胆碱能犯罪的强大抑制剂，导致肌肉 宫缩，癫痫发作和极端暴露。某些操作可以不可逆转地（几分钟之内）迅速 抑制ACHE，产生“老化”物种对氧化物反激活剂的难治性，例如2-丙氧母（2-PAM），这是 在美国的一部分OP治疗，最近对其效率进行了质疑。最后， 迫切需要发现新颖的方法来挽救ACHE活动，以减轻治疗 神经和肌肉作用有害。拟议研究的长期目标是发展 可行的，持久的急性和慢性OP暴露。以前为营救老年疼痛的努力 专注于产生将烷基化磷酸化酶烷基化的新反抗剂，允许2-PAM等氧气 恢复酶功能，但是这些化合物的缺点，例如血脑屏障不足 渗透性和体内效率有限。拟议的研究具有创新性，因为IT员工策略 旨在增加营业额老化的疼痛和新生细胞内疼痛的恢复。例如，抑制 或肌肉特异性蛋白激酶（MUSK）的丧失使ACHE复合物不稳定导致降解 酶。最近的研究表明，DOK7是ACHE复合物中的衔接蛋白，对于 麝香活性，更重要的是，在肌肉表面上定位。因此，当前的研究 目的是评估突触ACHE复合物中靶向蛋白的靶向是否可以增加疼痛的周转率，并且 恢复最佳的疼痛活性。追求这一目标的中心假设是抑制或退化 与老年疼痛相关的蛋白质会导致衰老酶的释放，使新的痛苦成为 重新填充突触，恢复最佳神经传递，并减轻OP暴露的影响。这 拟议的研究的理由是消除老化的疼痛将减轻OP的有害影响 毒性并恢复适当的神经传递。基于出版和初步研究，高中实习生 将调查父母赠款的特定目的1中的要素：评估靶向的体外治疗益处 ACHE复合物中的蛋白质降解。在拟议的项目中，高中实习生将使用小型RNA 降低DOK7的蛋白质水平。同样，学生将使用模仿DOK7-的复古式辣椒 - 麝香的相互作用位点破坏了蛋白质 - 蛋白质相互作用的稳定，我们假设这将导致释放 这些研究的年龄疼痛将确定DOK7的可药用性，并确定是否将其评估为药物 发现研究。这项研究很重要，因为它将为此提供基本的机械见解 稳定性和突触后的ACHE复合物和发掘新目标以治疗OP暴露。