Induced Therapeutic Overexpression of Acetylcholinesterase in Vivo

体内诱导乙酰胆碱酯酶的治疗性过度表达

基本信息

批准号：
8732706
负责人：
Richard Lee Rotundo
金额：
$ 52.33万
依托单位：
UNIVERSITY OF MIAMI SCHOOL OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-09-30 至 2016-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8732706
关键词：
Acetylcholinesterase Acetylcholinesterase Inhibitors Active Sites Animals Blood Circulation Brain Cessation of life Chimeric Proteins Cultured Cells Dependence Development Dose Enzymes Event Exposure to Funding Half-Life Home environment Hour In Vitro Laboratories Lead Lethal Dose 50 Life Ligands Longevity Methods Mus Muscle Nerve Nervous system structure Neuromuscular Junction Organophosphates Peptides Pesticides Pharmaceutical Preparations Principal Investigator Production Protein Biosynthesis Proteins Recovery Research Resistance Sarin Skeletal Muscle Soman Specificity Synapses Testing Therapeutic Time Variant base cholinergic cholinergic synapse efficacy testing improved in vivo inhibitor/antagonist mouse model nerve agent nerve gas neuromuscular neuromuscular transmission novel novel strategies overexpression prevent programs protective effect protein folding response small molecule tabun tissue culture transmission process

项目摘要

DESCRIPTION (provided by applicant): Acetylcholinesterase (AChE), the enzyme that terminates cholinergic transmission in the nervous system, is the target of organophosphate nerve agents and pesticides. Inactivation of AChE can result in rapid debilitation or death, thus the use of easily synthesized nerve agents such as soman and sarin by terrorists looms as a potentially catastrophic event. Our laboratory has recently shown that amino terminal peptides derived from the non-catalytic AChE targeting subunits stabilize the newly-synthesized AChE molecules and prevent their degradation in skeletal muscle. This in turn dramatically increases active enzyme and specifically AChE at the neuromuscular synapse when administered to living mice. This increase is rapid and sufficient to protect the animals from exposure to 2x LD50 DFP. Recent preliminary studies in our lab have uncovered small molecules that can enhance AChE folding, and an additional study has provided a novel mechanism for replacing damaged AChE at the neuromuscular synapse. Our specific aims on this project are; 1) to continue our studies on AChE-inducing/stabilizing peptides in vivo using our mouse model to optimize dose and delivery when administered before or after exposure to the surrogate nerve agent DFP; additional studies will test the efficacy of the PRAD-KDEL peptides against soman and tabun; 2) We have identified several ligands that can increase expression of active AChE by enhancing its folding; these compounds will be studied in cultured cells to determine their mechanism of action and test the hypothesis that they act in a manner analogous but not identical to the PRAD-KDEL peptides; possible synergies will be determined; 3) test the hypothesis that these ligands can increase AChE expression in vivo by enhancing folding and stabilization using our mouse model; and 4) Determine the ability of our novel AChE replacement approach using organophosphate resistant enzyme to protect mice from exposure to organophosphate nerve agents and their ability to improve survival when administered after exposure. Together these studies will lead to novel methods for increasing AChE expression in vivo for protection against organophosphate compounds as well as enhancing the rate of recovery following exposure. PUBLIC HEATLTH RELEVANCE: Acetylcholinesterase (AChE) is the enzyme that terminates neuromuscular transmission and is the target of organophosphate nerve gases and some pesticides. Our research is aimed at developing novel therapies to restore active AChE following its inactivation using a combination of peptides and drugs that increase the production of enzyme in the brain and muscles, as well as replacing damaged molecules with new ones.

描述（由申请人提供）：乙酰胆碱酯酶（AChE）是一种终止神经系统中胆碱能传递的酶，是有机磷神经毒剂和杀虫剂的目标。乙酰胆碱酯酶失活会导致迅速衰弱或死亡，因此恐怖分子使用容易合成的神经毒剂（例如索曼和沙林）可能会成为潜在的灾难性事件。我们的实验室最近表明，源自非催化 AChE 靶向亚基的氨基末端肽可以稳定新合成的 AChE 分子，并防止其在骨骼肌中降解。当给予活体小鼠时，这反过来会显着增加神经肌肉突触处的活性酶，特别是乙酰胆碱酯酶（AChE）。这种增加是快速且足以保护动物免于暴露于 2x LD50 DFP。我们实验室最近的初步研究发现了可以增强乙酰胆碱酯酶折叠的小分子，另外一项研究提供了一种新的机制来替代神经肌肉突触处受损的乙酰胆碱酯酶。我们该项目的具体目标是： 1) 使用我们的小鼠模型继续对乙酰胆碱酯酶诱导/稳定肽进行体内研究，以优化在接触替代神经毒剂 DFP 之前或之后给药时的剂量和递送；其他研究将测试 PRAD-KDEL 肽对抗梭曼和塔崩的功效； 2）我们已经鉴定出几种配体可以通过增强活性AChE的折叠来增加其表达；这些化合物将在培养细胞中进行研究，以确定其作用机制，并测试它们的作用方式与 PRAD-KDEL 肽类似但不相同的假设；将确定可能的协同效应； 3) 使用我们的小鼠模型测试这些配体可以通过增强折叠和稳定性来增加体内 AChE 表达的假设； 4) 确定我们的新型乙酰胆碱酯酶替代方法使用有机磷酸盐抗性酶保护小鼠免受有机磷酸盐神经毒剂暴露的能力，以及在接触后给药时提高存活率的能力。这些研究共同将带来增加体内 AChE 表达的新方法，以防止有机磷酸酯化合物的侵害，并提高暴露后的恢复率。公共卫生相关性：乙酰胆碱酯酶 (AChE) 是一种终止神经肌肉传递的酶，也是有机磷神经毒气和某些杀虫剂的目标。我们的研究旨在开发新的疗法，使用肽和药物的组合来恢复失活后的活性乙酰胆碱酯酶，以增加大脑和肌肉中酶的产生，以及用新的分子替换受损的分子。