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）：长期暴露于尼古丁是一种横切现象，导致尼古丁依赖性以及无意的治疗作用，例如保护帕金森氏病。该项目测试了新的建议，即慢性尼古丁暴露的影响取决于细胞内尼古丁对质膜上传统信号转导的传统信号转导。众所周知，尼古丁被动进入细胞，最近的研究表明尼古丁也进入细胞器，例如内质网（ER）。在ER中，尼古丁可以在药理学上伴侣新生的烟碱乙酰胆碱受体（NACHRS），作为五聚体受体组装的“对接会”亚基。其他研究表明，细胞内尼古丁 - NACHR相互作用的其他潜在后遗症：降低展开的蛋白质反应，“护送”其他蛋白质从ER或“绑扎”蛋白质到异常途径。 提出的机制是“内部”，因为它始于ER而不是质膜。尼古丁的持续内外效应以远低于其质膜NACHR通道的瞬时激活而进行。 该项目将发明新技术，以测量和控制“内部”烟碱药理学中的初始步骤。我们将开发用于测量药物结合的纳米菌素，并将开发分室化的烟碱配体。我们还将采用其他最先进的技术：Copii囊泡芽的重建和烦恼。 亚诉讼方法是测量尼古丁作用隔室化的工具。尼古丁和临床上重要的alpha4beta2 nachr选择性配体Varenicline的尼古丁和尼古丁的次接收工具用于将药理学限制在ER中。亚方法C检验了先前实验发现的NACHR和“候选基因”之间的相互作用。内而外的药理学是一个变革性的概念，可能揭示了成瘾和神经变性的新治疗靶标。