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Functional mechanisms of Hnrnph1 in methamphetamine addictive behaviors

Hnrnph1在甲基苯丙胺成瘾行为中的功能机制

基本信息

批准号：
9122756
负责人：
Neema Yazdani
金额：
$ 3.27万
依托单位：
BOSTON UNIVERSITY MEDICAL CAMPUS
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-04-01 至 2019-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9122756
关键词：
Acute Addictive Behavior Address Affect Alleles American Amphetamines Animal Model Behavior Behavioral Binding Cells Chromosomes, Human, Pair 11 Chronic Code Complex Congenic Mice Controlled Environment Corpus striatum structure Data Disease Dopamine Dopamine Receptor Dopaminergic Cell Dose Epidemiology FDA approved Functional disorder Genes Genetic Genetic Variation Heroin Dependence Human Human Genome Individual Differences Intervention Investigation Link Maps Measures Medical Mental disorders Messenger RNA Methamphetamine Methamphetamine dependence Microdialysis Midbrain structure Molecular Mus Neurons Oral Pathway Analysis Phenotype Physiological Prevention Property Proteins Psychological reinforcement Psychostimulant dependence Public Health Quantitative Trait Loci RNA Splicing RNA-Binding Proteins Regulation Reporting Rewards Risk Role Sample Size Self Administration Signal Transduction Staining method Stains Substance Use Disorder Testing Therapeutic Intervention Translations Tyrosine 3-Monooxygenase United States Variant Ventral Striatum addiction base body density congenic cost density dopaminergic neuron functional genomics gene repression genetic variant genome wide association study in vivo insight methamphetamine abuse mu opioid receptors multidisciplinary nerve supply neurobehavioral neuron development neuropsychiatric disorder neurotransmission novel preference presynaptic psychostimulant public health relevance resilience response targeted treatment trait transcription activator-like effector nucleases transcriptome transcriptome sequencing transmission process

项目摘要

DESCRIPTION (provided by applicant): Addiction to psychostimulants such as Methamphetamine (MA) is a significant public health issue in the United States and currently there are no FDA approved pharmacological interventions. Psychostimulant addiction is a heritable substance use disorder, however, its genetic basis is almost entirely unknown. Genome-wide association studies (GWAS) in humans currently lack sufficient power to detect the influence of common genetic variation on psychostimulant addiction. Mammalian model organisms offer an attractive alternative to more rapidly uncover novel genetic factors that contribute to addiction-relevant neurobehavioral traits. Using quantitative trait locus (QTL) mapping in mice, we identified a locus on chromosome 11 that caused a decrease in sensitivity to the locomotor stimulant properties of MA. To fine map this QTL, we generated interval-specific congenic lines and deduced a 206 kb critical interval on chromosome 11 that contained only two protein coding genes (Rufy1 and Hnrnph1). Replicate mouse lines heterozygous for Transcription Activator-like Effector Nucleases (TALENs)-induced frameshift deletions in Hnrnph1 (Hnrnph1+/-), but not in Rufy1 (Rufy1+/-), recapitulated the decrease in MA sensitivity observed in congenic mice, thus identifying Hnrnph1 as a novel quantitative trait gene for MA sensitivity. Hnrnph1 has not previously been identified in human GWAS of neuropsychiatric disorders but has been implicated in mu-opioid receptor splicing associated with heroin dependence. The primary objective of this proposal is to identify the functional mechanisms by which Hnrnph1 regulates MA addictive behaviors. RNA-sequencing and transcriptome analysis of the striatum suggests that inheritance of the QTL affects mesolimbic dopaminergic neuron development and striatal neurotransmission. Moreover, preliminary data extends the role of Hnrnph1 to the rewarding properties of MA as measured in the conditioned place preference (CPP) paradigm. To further investigate addiction-relevant behaviors in Hnrnph1+/- mice, in Aim 1 we will assess dose-dependent conditioned MA reward via MA-CPP as well as MA reinforcement via operant oral self-administration. In Aim 2, we will then identify the mesolimbic transcriptome, gene networks, spliceome, and the predicted HNRNPH1 targetome in the ventral midbrain and ventral striatum of Hnrnph1+/- mice. In Aim 3, we will use immunohistochemical staining of tyrosine hydroxylase to identify changes in ventral midbrain dopaminergic cell body density and presynaptic striatal innervation following heterozygous Hnrnph1 deletion. Additionally, we will determine the degree of co-localization between HNRNPH1 and D1 versus D2-type dopamine receptors in medium spiny neurons of the striatum. Finally, we will utilize in vivo microdialysis to measure baseline and MA-induced changes in striatal dopamine release in Hnrnph1+/- mice. These results will provide novel genomic and functional insights into Hnrnph1 regulation of mesolimibic circuitry function, MA addictive behaviors, and potentially inform upon novel targets for prevention and treatment of psychostimulant addiction.

描述（由申请人提供）：甲基苯丙胺（MA）等精神兴奋剂成瘾是美国的一个重大公共卫生问题，目前尚无FDA批准的药物干预。精神兴奋剂成瘾是一种遗传性物质使用障碍，然而，其遗传基础几乎完全未知。人类全基因组关联研究（GWAS）目前缺乏足够的能力来检测常见遗传变异对精神兴奋剂成瘾的影响。哺乳动物模式生物提供了一个有吸引力的替代更迅速地发现新的遗传因素，有助于成瘾相关的神经行为特征。利用小鼠数量性状基因座（QTL）定位，我们确定了11号染色体上的一个基因座，该基因座导致对MA运动刺激特性的敏感性降低。为了精细定位该QTL，我们产生了间隔特异性同源系，并在第11号染色体上推导出一个仅包含两个蛋白质编码基因（Rufy 1和Hnrnph 1）的206 kb临界间隔。转录激活因子样效应核酸酶（TALEN）诱导的Hnrnph 1（Hnrnph 1 +/-）而非Rufy 1（Rufy 1 +/-）移码缺失的杂合复制小鼠品系重现了在同源小鼠中观察到的MA敏感性降低，因此将Hnrnph 1鉴定为MA敏感性的新数量性状基因。Hnrnph 1以前未在神经精神疾病的人类GWAS中被鉴定，但与海洛因依赖相关的μ阿片受体剪接有关。该建议的主要目的是确定Hnrnph 1调节MA成瘾行为的功能机制。纹状体的RNA测序和转录组分析表明，QTL的遗传影响中脑边缘多巴胺能神经元发育和纹状体神经传递。此外，初步的数据扩展的作用Hnrnph 1的奖励性质的MA测量条件性位置偏好（CPP）的范例。为了进一步研究Hnrnph 1 +/-小鼠的成瘾相关行为，在目标1中，我们将评估通过MA-CPP的剂量依赖性条件MA奖励以及通过操作性口服自我给药的MA强化。在目标2中，我们将鉴定Hnrnph 1 +/-小鼠腹侧中脑和腹侧纹状体中的中脑边缘转录组、基因网络、剪接组和预测的HNRNPH 1靶组。在目标3中，我们将使用酪氨酸羟化酶的免疫组织化学染色，以确定腹侧中脑多巴胺能细胞体密度和突触前纹状体神经支配杂合Hnrnph 1缺失后的变化。此外，我们将确定HNRNPH 1和D1与D2型多巴胺受体在纹状体中棘神经元之间的共定位程度。最后，我们将利用体内微透析来测量Hnrnph 1 +/-小鼠纹状体多巴胺释放的基线和MA诱导的变化。这些结果将提供新的基因组和功能的见解Hnrnph 1调节mesolimibic电路功能，MA成瘾行为，并可能告知新的目标，预防和治疗精神兴奋剂成瘾。