Tools for inside-out pharmacology: nicotinic agents

由内而外的药理学工具：烟碱类药物

• 批准号: 8640727
• 负责人: Henry A. Lester
• 金额: $ 33.3万
• 依托单位: CALIFORNIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-01 至 2017-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8640727
• 关键词: Binding; Biological; Biological Assay; Biological Testing; Brain; Candidate Disease Gene; Cell Line; Cell membrane; Cells; Chemicals; Chronic; Cigarette; Collaborations; Detection; Drug Receptors; Drug effect disorder; Endoplasmic Reticulum; Equilibrium; Esters; Event; Exposure to; Family suidae; Farming environment; Fluorescence Resonance Energy Transfer; Functional disorder; Guidelines; Hour; Image; Kinetics; Knowledge; Learning; Life; Ligands; Masks; Measures; Membrane; Molecular; Molecular Chaperones; N-Methyl-D-Aspartate Receptors; Nerve Degeneration; Neurodegenerative Disorders; Neuropharmacology; Nicotine; Nicotine Dependence; Nicotinic Agents; Nicotinic Agonists; Nicotinic Receptors; Organelles; Osmosis; Parkinson Disease; Pathway interactions; Persons; Pharmaceutical Preparations; Pharmacology; Process; Proteins; Psychiatry; Receptor Protein-Tyrosine Kinases; Receptor Signaling; Research Personnel; Signal Transduction; Site; Spectrometry, Mass, Secondary Ion; Suggestion; System; Techniques; Testing; Therapeutic; Therapeutic Effect; Time; Tobacco; Up-Regulation; Vesicle; addiction; dopaminergic neuron; drug discovery; endoplasmic reticulum stress; esterase; heuristics; improved; nanoscale; new therapeutic target; novel; public health relevance; receptor; receptor binding; reconstitution; research study; response; theories; tool; varenicline

DESCRIPTION (provided by applicant): Chronic exposure to nicotine is a crosscutting phenomenon, leading to nicotine dependence as well as to inadvertent therapeutic effects such as protection against Parkinson's disease. The project tests the novel suggestion that effects of chronic nicotine exposure depend on intracellular nicotine-not on conventional signal transduction via receptors at the plasma membrane. It is known that nicotine passively enters cells and recent studies suggest that nicotine also enters organelles such as endoplasmic reticulum (ER). In ER, nicotine may pharmacologically chaperone nascent nicotinic acetylcholine receptors (nAChRs), "matchmaking" subunits as pentameric receptors assemble. Other studies suggest additional potential sequelae of intracellular nicotine-nAChR interactions: decreasing unfolded protein responses, "escorting" other proteins from ER, or "abducting" proteins to abnormal pathways. The proposed mechanism is "inside-out", because it begins in the ER rather than on the plasma membrane. Nicotine's sustained inside-out effects proceed at concentrations much lower than its transient activation of plasma membrane nAChR channels. The project will invent new techniques to measure and control the initial steps in "inside-out" nicotinic pharmacology. We will develop NanoSIMS for measuring drug binding, and we will develop compartmentalized nicotinic ligands. We will also employ other state-of-the-art techniques: reconstitution of COPII vesicle budding, and FRET. Sub-Approach A invents tools measuring the compartmentalization of nicotine action. Sub-Approach B invents tools for confining pharmacology to the ER, both for nicotine and for the clinically important alpha4beta2 nAChR-selective ligand, varenicline. Sub-Approach C tests for interactions between nAChRs and "candidate genes" discovered by previous experiments. Inside-out pharmacology is a transformative concept that may reveal new therapeutic targets for addiction and neurodegeneration.

描述（由申请人提供）：长期接触尼古丁是一种交叉现象，会导致尼古丁依赖以及无意的治疗效果，例如预防帕金森病。该项目测试了一个新的建议，即长期接触尼古丁的影响取决于细胞内的尼古丁，而不是通过质膜受体的传统信号转导。众所周知，尼古丁被动地进入细胞，最近的研究表明尼古丁还可以进入内质网（ER）等细胞器。在 ER 中，尼古丁可以在药理学上陪伴新生烟碱乙酰胆碱受体 (nAChR)，即五聚体受体组装时的“配对”亚基。其他研究表明细胞内尼古丁 - nAChR 相互作用还可能产生其他潜在后遗症：减少未折叠蛋白反应，“护送”内质网中的其他蛋白，或将蛋白“诱拐”至异常途径。 所提出的机制是“由内而外”的，因为它始于内质网而不是质膜。尼古丁持续由内而外的作用在远低于其质膜 nAChR 通道瞬时激活的浓度下进行。 该项目将发明新技术来测量和控制“由内而外”烟碱药理学的初始步骤。我们将开发用于测量药物结合的 NanoSIMS，并且我们将开发区室化烟碱配体。我们还将采用其他最先进的技术：COPII 囊泡出芽的重建和 FRET。 子方法 A 发明了测量尼古丁作用分区的工具。子方法 B 发明了将药理学限制在 ER 内的工具，包括尼古丁和临床上重要的 α4β2 nAChR 选择性配体伐尼克兰。子方法 C 测试 nAChR 和先前实验发现的“候选基因”之间的相互作用。由内而外的药理学是一个变革性的概念，可能会揭示成瘾和神经退行性疾病的新治疗靶点。