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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

基本信息

批准号：
10322460
负责人：
Rajani Maiya
金额：
$ 32.61万
依托单位：
LSU HEALTH SCIENCES CENTER
依托单位国家：
美国
项目类别：
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/10322460
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 gene network 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

项目摘要

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.

项目总结