Variants in CHRNA5/CHRNA3/CHRNB4 and nicotine dependence

CHRNA5/CHRNA3/CHRNB4 变异与尼古丁依赖

• 批准号: 8133300
• 负责人: XIANGNING CHEN
• 金额: $ 3.89万
• 依托单位: VIRGINIA COMMONWEALTH UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2013-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8133300
• 关键词: 15q25; Acute; Affect; Agonist; Alcoholism; Amino Acids; Animal Model; Animals; Anxiety; Asparagine; Behavior; Behavioral; Binding; Biological Assay; Biological Models; Bupropion; Candidate Disease Gene; Cell Line; Cell model; Cells; Cholinergic Receptors; Chromosomes; Cigarette; Cocaine Dependence; Code; Comparative Study; Complex; Cultured Cells; D-Aspartic Acid; Data; Dependence; Development; Discrimination; Environmental Exposure; Environmental Risk Factor; Epithelial Cells; Equation; Evaluation; Follow-Up Studies; Foundations; Future; Genes; Genetic; Genetic Polymorphism; Genotype; Goals; Human; Immunoprecipitation; In Vitro; Knock-in Mouse; Knockout Mice; Lead; Ligands; Malignant neoplasm of lung; Measures; Mecamylamine; Modeling; Molecular; Motor Activity; Nicotine; Nicotine Dependence; Nicotinic Receptors; Patch-Clamp Techniques; Pharmaceutical Preparations; Physiologic pulse; Play; Property; Public Health; Rewards; Risk; Role; Single Nucleotide Polymorphism; Smoke; Smoking; Smoking Behavior; Structure; Syndrome; Techniques; Testing; Time; Tobacco smoking; Transgenic Organisms; Twin Multiple Birth; Variant; acetylcholine receptor agonist; addiction; base; behavior influence; desensitization; design; epidemiology study; established cell line; gene environment interaction; genetic variant; patch clamp; preference; public health relevance; radioligand; receptor; receptor expression; response; varenicline

DESCRIPTION (provided by applicant): Tobacco smoking is addictive and thought to be driven by nicotine dependence (ND). For decades, data have suggested genetic bases for an increased risk of ND. More recently, several studies validated the association of rs16969968 with nicotine dependence. rs16969968 is a non-synonymous SNP, changing the 398th amino acid from aspartic acid (D) to asparagine (N) (D398N) in the CHRNA5 gene. We hypothesize that this non-synonymous variant in the CHRNA5 gene may alter the functional properties of nAChRs containing 5 subunit, ultimately altering risks for tobacco smoking and ND. To test this hypothesis, the project has four aims: 1. To generate stably transfected cell lines expressing human a4b2*- or a3b4*-nAChRs containing wild- type or variant a5 subunits and use these cell lines as model system to study the properties of major subtypes (a4b2* and a3b4*) in vitro. 2. To characterize functional and pharmacological effects of the presence of wild-type or D398N a5 subunits on a4b2*- and a3b4*-nAChR using patch-clamp technique. 3. To test whether rs16969968 is involved in smoking initiation and to test for gene-environment interactions -that is, to determine if the impact of environmental exposures on smoking behaviors are modified by the rs16969968 genotypes. 4. To establish roles of a5-containing nAChR subtypes in ND using a5 knock-out mice. The goal is to define how the presence or absence of a5 subunits in native nAChR affects acute responses to nicotine (anxiety and locomotor activity), nicotine reward (conditioned place preference, CPP), and subjective effects (nicotine discrimination). PUBLIC HEALTH RELEVANCE: Tobacco smoking and nicotine dependence is a major public health issue. Many epidemiology studies have provided compelling evidence that genetic factors play a significant role in the development of nicotine dependence. More recently, several large scale genetic studies have found that several variants in the nicotinic receptor alpha 5, alpha 3 and beta 4 genes are significantly associated with heavy smoking and nicotine dependence, including one functional polymorphism rs16969968 or D398N. This current study is designed to create model systems to examine the effects of D398N at the levels of cultured cells, animal model and human behaviors. The approaches we take are 1) To develop a cellular model system to study molecular, pharmacological and electrophysiological properties of N398D; 2) To examine how genetic variants at rs16969968 interact with environmental factors and eventually lead to smoking addiction and dependence; and 3) To study the behavioral effects of these genes in animal models.

描述（由申请人提供）：吸烟会让人上瘾，并且被认为是由尼古丁依赖（ND）驱动的。几十年来，数据表明新城疫风险增加存在遗传基础。最近，几项研究验证了 rs16969968 与尼古丁依赖的关联。 rs16969968 是一个非同义 SNP，将 CHRNA5 基因中的第 398 个氨基酸从天冬氨酸 (D) 更改为天冬酰胺 (N) (D398N)。我们假设 CHRNA5 基因中的这种非同义变异可能会改变包含 5 个亚基的 nAChR 的功能特性，最终改变吸烟和 ND 的风险。为了检验这一假设，该项目有四个目标： 1. 生成表达含有野生型或变异型 a5 亚基的人 a4b2*- 或 a3b4*-nAChR 的稳定转染细胞系，并使用这些细胞系作为模型系统在体外研究主要亚型（a4b2* 和 a3b4*）的特性。 2. 使用膜片钳技术表征野生型或 D398N a5 亚基的存在对 a4b2*- 和 a3b4*-nAChR 的功能和药理作用。 3. 测试rs16969968是否参与吸烟启动并测试基因-环境相互作用，即确定环境暴露对吸烟行为的影响是否被rs16969968基因型改变。 4.使用a5敲除小鼠确定含有a5的nAChR亚型在ND中的作用。目标是确定天然 nAChR 中 a5 亚基的存在或缺失如何影响对尼古丁的急性反应（焦虑和运动活动）、尼古丁奖励（条件位置偏好，CPP）和主观效应（尼古丁歧视）。 公共卫生相关性：吸烟和尼古丁依赖是一个主要的公共卫生问题。许多流行病学研究提供了令人信服的证据，证明遗传因素在尼古丁依赖的发展中发挥着重要作用。最近，几项大规模遗传学研究发现烟碱受体α5、α3和β4基因中的几种变异与重度吸烟和尼古丁依赖显着相关，其中包括一种功能多态性rs16969968或D398N。目前的研究旨在创建模型系统来检查 D398N 在培养细胞、动物模型和人类行为水平上的影响。我们采取的方法是1）开发细胞模型系统来研究N398D的分子、药理学和电生理学特性； 2) 研究rs16969968基因变异如何与环境因素相互作用并最终导致吸烟成瘾和依赖； 3) 研究这些基因在动物模型中的行为影响。

项目成果

Controlled drug-release system based on pH-sensitive chloride-triggerable liposomes.

基于 pH 敏感氯化物可触发脂质体的受控药物释放系统。

• DOI: 10.3109/08982104.2012.727423
• 发表时间: 2013
• 期刊: Journal of liposome research
• 影响因子: 4.4
• 作者: Wehunt,MarkP;Winschel,ChristineA;Khan,AliK;Guo,TaiL;Abdrakhmanova,GalyaR;Sidorov,Vladimir
• 通讯作者: Sidorov,Vladimir