Excessive Alcohol Drinking Associated with GABA Alpha 2-Regulated TLR4 Expression

过量饮酒与 GABA Alpha 2 调节的 TLR4 表达相关

• 批准号: 8439773
• 负责人: Laure Aurelian
• 金额: $ 39.08万
• 依托单位: HOWARD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-06-20 至 2018-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8439773
• 关键词: Abstinence; Adult; Alcohol consumption; Alcohol dependence; Alcohols; Amygdaloid structure; Brain; Communication; DSM-V; Data; Dopamine; Enzymes; GABA Receptor; Gene Targeting; Genes; Globus Pallidus; Goals; Grant; Health; Heavy Drinking; Hour; Measures; Mediating; Modeling; Monocyte Chemoattractant Protein-1; Natural Immunity; Neurons; Neurosciences; Neurotransmitters; Play; Public Health; Rattus; Regulation; Relapse; Reporting; Research; Rewards; Role; Schedule; Signal Transduction; Simplexvirus; Site; Small Interfering RNA; Testing; Tyrosine 3-Monooxygenase; Work; alcohol related problem; alcohol reward; attenuation; base; binge drinking; cell type; chemokine; defined contribution; deprivation; dopaminergic neuron; drinking; epidemiologic data; gamma-Aminobutyric Acid; gene therapy; innovation; meetings; neurotransmission; problem drinker; public health relevance; receptor; response; therapeutic gene; toll-like receptor 4; transcription factor; vector

DESCRIPTION (provided by applicant): Excessive alcohol drinking is an enormous public health burden in need of more radical therapy. Most health problems associated with excessive drinking are due to two types of drinking: (i) binge (BALs > 0.08 grams % in a 2-hour period) and (ii) relapse (sustained heavy drinking for at least 2 days following a single or multiple abstinence periods). Previously, we showed that alcohol- preferring (P) rats express elevated levels of the GABAA subunits for alpha1, alpha2 and toll-like receptor 4 (TLR4) innate immunity receptors, and using specific siRNA vectors infused into the central amygdala (CeA), demonstrated that regulation of binge drinking at this site is mediated by GABAA a2 regulated TLR4 (a2/TLR4 axis). While the alpha1 subunit was also shown to regulate binge drinking, it was associated with the ventral pallidum (VP) and was independent of TLR4 (Liu et al., PNAS, 2011). In the current grant, we propose to better elucidate the mechanism responsible for the TLR4 effect, focusing on the chemokine monocyte chemotactic protein-1 (MCP-1) and the dopamine rate- limiting enzyme tyrosine hydroxylase (TH)--implicated by data obtained after the application was last submitted--and on brain sites that regulate different domains of the alcohol addiction cycle. The working hypothesis is that neuronal TLR4 induces MCP-1 expression via activation of cell type- specific transcription factors and it, in turn, functions as a neurotransmitter to stimulate dopamine release and excitability (inferred by TH) in select reward loci. The specific aims are: Aim I. Define the expression of GABAA a2/a1, TLR4, MCP-1 and/or TH at alcohol reward loci from P vs NP rats. Aim II. Define the role played by the a2/TLR4 axis that encompasses MCP-1 and/TH at alcohol reward loci in impulsive binge drinking. Aim III. Define the contribution of a1, a2, TLR4 and its downstream targets (MCP-1 and/or TH) at alcohol reward loci in compulsive relapse drinking. Aim IV. Define the mechanism of TLR4-mediated regulation of binge drinking by focusing on downstream signals that upregulate MCP-1 and TH expression, through the use of the pHSVsiMCP-1 amplicon. Better understanding of the role of chemokines in neurotransmission and the regulation of distinct (viz. dopaminergic) neurons will help broaden current concepts of neuroimmune communication and their roles in excessive drinking. This highly innovative proposal will create a paradigm shift in understanding the relationship between innate immunity signals and neuronal responses that impact alcohol addiction. Use of non-toxic herpes simplex virus (HSV)- siRNA constructs to inhibit relevant genes at specific brain loci will define therapeutic gene targets and test the potential of gene therapy approaches for binge and relapse drinking.

描述（由申请人提供）：过度饮酒是一个巨大的公共卫生负担，需要更彻底的治疗。与过量饮酒相关的大多数健康问题是由于两种类型的饮酒：（i）狂饮（2小时内巴尔斯> 0.08克%）和（ii）复发（在单次或多次戒酒后持续大量饮酒至少2天）。先前，我们显示了酒精偏好（P）大鼠表达升高水平的α 1、α 2和toll样受体4（TLR 4）先天免疫受体的GABAA亚基，并且使用输注到中央杏仁核（CeA）中的特异性siRNA载体，证明了在该位点的狂饮的调节是由GABAA α 2调节的TLR 4（α 2/TLR 4轴）介导的。虽然alpha 1亚基也显示出调节狂饮，但它与腹侧苍白球（VP）相关，并且不依赖于TLR 4（Liu et al.，PNAS，2011）。在目前的资助中，我们建议更好地阐明负责TLR 4效应的机制，重点关注趋化因子单核细胞趋化蛋白-1（MCP-1）和多巴胺限速酶酪氨酸羟化酶（TH）-由上次提交申请后获得的数据所暗示-以及调节酒精成瘾循环不同领域的大脑部位。工作假设是神经元TLR 4通过激活细胞类型特异性转录因子诱导MCP-1表达，并且其反过来作为神经递质在选择的奖赏基因座中刺激多巴胺释放和兴奋性（由TH推断）。具体目标是：确定P与NP大鼠酒精奖励位点GABAA a2/a1、TLR 4、MCP-1和/或TH的表达。Aim II.定义α 2/TLR 4轴在冲动性酗酒中酒精奖励位点所起的作用，该轴包括MCP-1和/TH。Aim III.定义a1，a2，TLR 4及其下游靶点（MCP-1和/或TH）在强迫性复吸饮酒中酒精奖励位点的作用。目标四。通过使用pHSVsiMCP-1扩增子，通过关注上调MCP-1和TH表达的下游信号，定义TLR 4介导的酗酒调节机制。更好地理解趋化因子在神经传递中的作用和不同（即多巴胺能）神经元的调节将有助于拓宽目前神经免疫通讯的概念及其在过度饮酒中的作用。这一高度创新的提议将在理解影响酒精成瘾的先天免疫信号和神经元反应之间的关系方面创造一个范式转变。使用无毒的单纯疱疹病毒（HSV）- siRNA结构来抑制特定脑位点的相关基因，将确定治疗基因靶点，并测试基因治疗方法对酗酒和复发性饮酒的潜力。