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

• 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
• 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

 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（由适用提供）：对甲基苯丙胺（MA）等心理刺激剂的成瘾是美国的重要公共卫生问题，目前尚无FDA批准的药物干预措施。心理刺激成瘾是一种可遗传的药物使用障碍，但是，其遗传基础几乎完全未知。目前，全基因组关联研究（GWAS）缺乏足够的能力来检测常见遗传变异对精神刺激成瘾的影响。哺乳动物模型生物提供了一种有吸引力的替代方法，可探索更迅速的新型遗传因素，这些因素有助于成瘾的神经行为特征。使用小鼠中的定量性状基因座（QTL）映射，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的运动刺激特性的敏感性降低。为了仔细映射此QTL，我们生成了间隔特异性的先天性线，并推导了11个染色体上的206 kb临界区间，该染色体仅包含两个蛋白质编码基因（Rufy1和HnRNPH1）。复制小鼠系在HNRNPH1（HNRNPH1 +/-）中的转录活化剂样效应子核酸核酸酶（TALES）诱导的移码缺失，但在Rufy1（Rufy1 +/-）中却不是在MA的敏感性中，因此在Rufy1（Rufy1 +/-）中均可降低。灵敏度。 HNRNPH1以前尚未在神经精神疾病的人类GWA中鉴定出来，但在与海洛因依赖相关的MU-阿片类受体剪接中隐含。该提案的主要目的是确定HNRNPH1调节MA添加剂行为的功能机制。纹状体的RNA测序和转录组分析表明，QTL的遗传会影响中唇多巴胺能神经发育和纹状体神经传递。此外，初步数据将HNRNPH1的作用扩展到MA的奖励性能，如条件的位置偏好（CPP）范式所测量。为了进一步研究HNRNPH1 +/-小鼠中与成瘾相关的行为，在AIM 1中，我们将通过MA-CPP以及通过操作口服自我给药来评估依赖剂量的条件MA奖励以及MA增强。然后，在AIM 2中，我们将确定Hnrnph1 +/-小鼠的腹脑中脑和腹侧纹状体中的中唇转录组，基因网络，剪接组和预测的HNRNPH1靶体。在AIM 3中，我们将使用酪氨酸羟化酶的免疫组织化学染色来鉴定杂合HNRNPH1缺失后腹中脑中多巴胺能细胞体密度和突触前纹状体神经支配的变化。此外，我们将确定纹状体中培养基神经元中HnRNPH1和D1与D2型多巴胺受体之间的共定位程度。最后，我们将利用体内微透析来测量HNRNPH1 +/-小鼠中纹状体多巴胺释放的基线和MA诱导的变化。这些结果将为HNRNPH1调节中溶膜电路功能，MA附加行为提供新的基因组和功能见解，并有可能告知预防和治疗精神刺激成瘾的新目标。