Genetic Influences Over Nicotine Withdrawal

遗传对尼古丁戒断的影响

• 批准号: 8144801
• 负责人: Mariella De Biasi
• 金额: $ 18.98万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-17 至 2013-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8144801
• 关键词: Abstinence; Acute; Affective; Alleles; Amino Acids; Animals; Anxiety; Area; Asparagine; Aspartic Acid; Behavior; Behavioral; Biological Factors; Brain; Brain region; Cells; Characteristics; Cigarette; Conotoxin; DSM-IV; Data; Dependence; Development; Frequencies; Gene Cluster; Genes; Genetic; Genetic Techniques; Genetic Variation; Genetically Engineered Mouse; Habenula; Health; Hippocampus (Brain); Individual; Injection of therapeutic agent; Kinetics; Knockout Mice; Laboratories; Lead; Lentivirus Vector; Linkage Disequilibrium; Mecamylamine; Medial; Medical; Microinjections; Molecular; Molecular Biology; Mus; Neurons; Nicotine; Nicotine Dependence; Nicotine Withdrawal; Nicotinic Receptors; Patch-Clamp Techniques; Pharmacogenetics; Pharmacology; Positioning Attribute; Property; Protein Isoforms; Receptor Gene; Reporting; Resistance; Risk; Role; Saccharin; Sampling; Severities; Single Nucleotide Polymorphism; Slice; Smoker; Smoking; Social Problems; Subfamily lentivirinae; Substance Withdrawal Syndrome; Symptoms; System; Tobacco; Tobacco use; Translating; Twin Multiple Birth; Variant; Ventral Tegmental Area; Wild Type Mouse; Withdrawal; Withdrawal Symptom; World Health Organization; acetylcholine receptor agonist; base; cigarette smoking; desensitization; drinking water; epibatidine; genetic manipulation; genetic variant; genome wide association study; in vivo; interest; interpeduncular nucleus; knock-down; novel; null mutation; particle; patch clamp; public health relevance; research study; response; selective expression; smoking cessation; success

DESCRIPTION (provided by applicant): According to the World Health Organization, about 10 million cigarettes are sold every minute in the world and every eight seconds someone dies from tobacco use (WHO, 2002). Nicotine is the main addictive component of tobacco, and the affective and somatic symptoms associated with nicotine withdrawal contribute to the difficulty in quitting tobacco use. While current therapies for smoking cessation are helpful, none can claim a very high rate of success. Therefore, there is a great need for a better understanding of the behavioral and biological factors underlying nicotine withdrawal. The persistence of cigarette smoking is influenced by genetic factors as highlighted by the recently found correlation between nicotine dependence and single nucleotide polymorphism (SNP) in the CHRNA5-CHRNA3-CHRNB4 nAChR gene cluster. This application builds on the observation that lack of ?5-containing nicotinic acetylcholine receptors (?5* nAChRs) abolishes somatic signs of withdrawal and reduces anxiety-like responses. ?5* nAChRs are expressed in the medial habenula (MHb) and interpeduncular nucleus (IPN). Recent studies from the lab have shown that injection of the nAChR antagonist, mecamylamine, in either the MHb or IPN is sufficient to precipitate withdrawal signs in nicotine-treated mice. We will first ask whether the rs16969968 SNP, which causes a D398N aminoacid change, can alter the biophysical and pharmacological properties of the ?5* nAChR subtypes expressed in MHb and IPN. Patch clamp experiments will be conducted in heterologous expression systems on nAChRs comprising the D398 or N398 ?5 subunit variants. Second, to investigate the direct role of ?5* nAChRs in MHb and IPN, lentiviral vectors will be used to either express ?5 gene variants in the MHb and IPN of ?5 null mice or to knock down the levels of endogenous ?5 in the MHb and IPN of wild type mice. MHb and IPN slices from those mice will be used in patch clamping experiments to determine the effect of the genetic manipulations on nicotine-induced increases in spike firing frequency. In other experiments, lentivirus-injected mice will be exposed to nicotine in the drinking water for six weeks to study mecamylamine-precipitated withdrawal. The studies will determine whether expression of ?5* nAChRs is necessary and sufficient for the manifestation of nicotine withdrawal and whether the potential molecular changes produced by the rs16969968 15 SNP translate into different withdrawal symptoms in vivo. PUBLIC HEALTH RELEVANCE: Our studies will take advantage of genetically engineered mice and novel genetic techniques, and will combine behavioral analysis, molecular biology, and pharmacology to study the influence of an identified single nucleotide polymorphism on the manifestations of nicotine withdrawal. The studies could lead to the development of personalized smoking cessation therapies.

描述（由申请人提供）：根据世界卫生组织，世界卫生组织每分钟每分钟每分钟出售约1000万支香烟，每八秒钟有人死于烟草使用（WHO，2002年）。尼古丁是烟草的主要成瘾成分，与尼古丁戒断相关的情感症状和躯体症状有助于戒烟的难度。尽管目前的戒烟疗法有帮助，但没有人可以夺得很高的成功率。因此，非常需要更好地了解尼古丁戒断的行为和生物学因素。 吸烟的持久性受遗传因素的影响，这是由ChRNA5-CHRNA3-CHRNB4 NACHR基因簇中最近发现的尼古丁依赖性与单核苷酸多态性（SNP）之间的相关性所突出的。该应用是基于缺乏5个含5个含5个烟碱的乙酰胆碱受体（？5* nACHR）的观察结果，它消除了戒断的躯体迹象并减少了焦虑样反应。 ？5* NACHR在内侧Habenula（MHB）和枝间核（IPN）中表达。实验室的最新研究表明，在MHB或IPN中注射NACHR拮抗剂Mecamylamine足以沉淀经尼古丁处理的小鼠的戒断体征。 我们将首先询问导致D398N氨基酸变化的RS16969968 SNP是否可以改变MHB和IPN中表达的？5* NACHR子类型的生物物理和药理特性。斑块夹实验将在包含D398或N398？5个亚基变体的NACHR的异源表达系统中进行。其次，为了调查？5* nACHR在MHB和IPN中的直接作用，慢病毒向量将用于表达5个无效小鼠的MHB和IPN中的5个基因变体，或者降低内源性？5的水平？野生型小鼠的MHB和IPN。这些小鼠的MHB和IPN切片将用于贴片夹具实验中，以确定遗传操纵对尼古丁引起的峰值发射频率增加的影响。在其他实验中，注射慢病毒的小鼠将在饮用水中暴露于尼古丁六个星期，以研究甲基胺预测的戒断。这些研究将确定nachrs的表达是否需要且足以表现出尼古丁戒断的表现，以及rs16969968 rs1696968产生的潜在分子变化是否在体内转化为不同的戒断症状。 公共卫生相关性：我们的研究将利用基因工程的小鼠和新颖的遗传技术，并将行为分析，分子生物学和药理学相结合，以研究确定的单核苷酸多态性对尼古丁戒断表现的影响。这些研究可能导致个性化戒烟疗法的发展。