Role of the transcriptional regulator Lmo4 in alcohol consumption and reward

转录调节因子 Lmo4 在饮酒和奖励中的作用

• 批准号: 10268581
• 负责人: Rajani Maiya
• 金额: $ 29.35万
• 依托单位: LSU HEALTH SCIENCES CENTER
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-01-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10268581
• 关键词: Address; Affinity; Affinity Chromatography; Alcohol consumption; Alcohol dependence; Alcohols; Amygdaloid structure; Binding; Brain; Candidate Disease Gene; Cues; Dependence; Desire for food; Development; Disintegrins; Dopamine; Down-Regulation; Ethanol; Event; Extracellular Matrix; Fright; Gene Expression; Gene Expression Profiling; Genes; Genetic; Genetic Transcription; Goals; Heavy Drinking; Heparitin Sulfate; Human; Knowledge; Lead; Learning; Lentivirus; Mediating; Messenger RNA; Metalloproteases; Molecular; Motivation; Mus; Neurons; Ontology; Pathway Analysis; Pathway interactions; Phenocopy; Procedures; Process; RNA Interference; Regulation; Rewards; Rodent; Role; Structure; Sulfatases; System; Testing; Therapeutic; Thrombospondins; Transcript; Viral; Work; alcohol exposure; alcohol reward; alcohol use disorder; base; cell type; chromatin immunoprecipitation; conditioned place preference; density; differential expression; drinking; drug reward; experimental study; hippocampal pyramidal neuron; kappa opioid receptors; knock-down; new therapeutic target; novel; preference; problem drinker; promoter; recruit; relating to nervous system; response; small hairpin RNA; transcription factor; transcriptome; transcriptome sequencing

Project Summary Repeated exposure to alcohol leads to neuroadapative changes that underlie the transition from moderate to excessive alcohol intake. Gene expression profiling studies in human alcoholics and rodents have led to the identification of a multitude of ethanol-responsive gene networks and pathways. However, there is a gap in knowledge in our understanding of how these networks are coordinated into a neuroadaptive response. One potential mechanism could involve the recruitment of transcription factors and transcriptional co-regulators that could modulate the expression of several downstream genes. However, very few studies have examined the role of transcriptional co-regulators in alcohol use disorders. Our preliminary results implicate a novel role for the transcriptional co-regulator Lim-Only 4 (LMO4) in regulating alcohol intake. LMO4 knockdown in the basolateral amygdala (BLA) led to a significant decrease in alcohol consumption in the intermittent access procedure and a significant deficit in conditioned place preference to alcohol suggesting a role for LMO4 in the BLA in regulating both alcohol consumption and reward. Unbiased transcriptome analysis of the BLA from WT and Lmo4gt/+ mice, which make 50% less Lmo4 than WT mice, using RNASeq revealed several genes that were differentially expressed including the kappa opioid receptor (Oprk1). Weighted gene co-expression network analysis (WGCNA) revealed extracellular matrix (ECM)-related genes as being dysregulated upon LMO4 knockdown. These results led us to hypothesize that a LMO4-regulated transcriptional network in the BLA regulates alcohol consumption. We propose the following specific aims to test this hypothesis. In Aim 1, we will determine whether Oprk1 functions downstream of LMO4 in the BLA to regulate alcohol consumption. We will use a combination of approaches including cell-type specific shRNA-mediated knockdown of Lmo4 and Oprk1 in BLA pyramidal neurons and chromatin immunoprecipitation to address this question. In Aim 2, we will determine whether LMO4 downregulation in the BLA causes abnormalities in the density of perineuronal nets (PNNs), a highly specialized form of ECM in the brain. We will next determine whether enzymatic dissolution of PNNs will reduce ethanol consumption. We will also determine whether a disintegrin and metalloproteinase with thrombospondin motif (ADAMTS) 2 and sulfatase 2 (Sulf2) function downstream of LMO4 to regulate alcohol consumption. Finally, in Aim 3, we will use a viral-based translational affinity purification strategy to determine how the LMO4-regulated transcriptome changes with alcohol exposure. The proposed experiments are significant because identification of transcriptional targets functioning downstream of LMO4 to regulate alcohol consumption could lead to the identification of novel therapeutic targets to treat alcohol use disorders.

项目摘要 反复接触酒精会导致神经自传变化，这是从中等转变到的转变 过多的酒精摄入量。人类酗酒和啮齿动物中的基因表达分析研究已导致 鉴定多种乙醇响应基因网络和途径。但是，存在差距 了解这些网络如何协调为神经适应性反应时的知识。一 潜在的机制可能涉及转录因子和转录共同调节因子的募集 可以调节几个下游基因的表达。但是，很少有研究检查 转录共同调节因子在酒精使用障碍中的作用。我们的初步结果暗示了 转录共同调节器仅限4（LMO4）调节酒精摄入量。 LMO4敲低 基底外侧杏仁核（BLA）导致间歇性进入的酒精消耗量显着降低 在条件地点偏爱酒精的过程中，有明显的赤字表明LMO4在 在调节酒精消耗和奖励方面的bla。 WT的BLA的无偏转录组分析 使用RNASEQ揭示了几个基因，其LMO4GT/+小鼠的LMO4比WT小鼠少50％。 差异表达，包括Kappa阿片受体（OPRK1）。加权基因共表达 网络分析（WGCNA）揭示了细胞外基质（ECM）相关的基因，因为在 LMO4敲低。这些结果使我们假设在 BLA调节饮酒。我们提出以下特定旨在检验该假设的特定旨在。在AIM 1中， 我们将确定OPRK1在BLA中下游是否在调节酒精消耗的情况下功能。 我们将使用多种方法组合 BLA锥体神经元和染色质免疫沉淀中的OPRK1解决了这个问题。在AIM 2中，我们将 确定BLA中的LMO4下调是否会导致会阴网密度的异常 （PNNS），一种高度专业的ECM，大脑中的ECM。接下来，我们将确定酶促溶解是否 PNNS将减少乙醇消耗。我们还将确定崩解蛋白和金属蛋白酶是否 LMO4下游的血小板传播基序（ADAMTS）2（ADAMTS）2（Sulf2）功能调节 饮酒。最后，在AIM 3中，我们将使用基于病毒的翻译亲和力净化策略来 确定LMO4调节的转录组如何随着酒精暴露而变化。提出的实验 之所以重要，是因为识别LMO4下游功能的转录靶标并调节 饮酒可能会导致鉴定出新的治疗靶标，以治疗酒精使用障碍。