Gene expression regulatory circuitry and microRNAs in midbrain dopamine neurons

中脑多巴胺神经元中的基因表达调控电路和 microRNA

• 批准号: 7572393
• 负责人: Asa Abeliovich
• 金额: $ 34.77万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-12-18 至 2013-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7572393
• 关键词: Adult; Autistic Disorder; Behavior; Biological Models; Brain Diseases; Cell Culture Techniques; Cell Differentiation process; Cell Survival; Cell physiology; Code; Complex; Data; Development; Disease; Dopamine; Drug Addiction; Dyskinetic syndrome; Enzymes; Feedback; Gene Expression; Gene Expression Regulation; Genes; Knockout Mice; Manuscripts; MicroRNAs; Midbrain structure; Modeling; Molecular; Molecular Analysis; Molecular Profiling; Mutant Strains Mice; Mutate; Neurons; Nucleotides; Parkinson Disease; Pathway interactions; Phenotype; Play; Post-Transcriptional Regulation; Process; Proteins; Publishing; RNA; Regulation; Rodent; Role; Schizophrenia; Signal Transduction; Technology; Testing; Tissues; Transcript; Untranslated Regions; Viral; aphakia mice; base; dopaminergic neuron; embryonic stem cell; human DICER1 protein; in vivo; neuron development; overexpression; postnatal; progenitor; public health relevance; transcription factor

DESCRIPTION (provided by applicant): A number of developmental and adult brain disorders are associated with midbrain dopamine neurons (mDNs), including Parkinson's disease (PD), schizophrenia, autism, dyskinesias, and drug addiction. Thus, the fundamental mechanisms that regulate the development and function of these cells are of great import. In prior studies, we and others have investigated basic regulatory processes, such as regulation of the expression of key enzymes in the dopamine biosynthetic pathway in the context of dopamine neuron development, function, and survival. However, it is clear from these studies that there is a high level of complexity governing all of these processes. For instance, at least 2 transcription factors, Nurr1 and Pitx3, function synergistically to govern expression of late developmental mDN markers in simplified ES cell-based culture models. More recently, in preliminary data and a published manuscript, my lab has found evidence of an added layer of complexity involving post-transcriptional regulation by microRNAs (miRNAs) in the context of mDN development and function. miRNAs are evolutionarily conserved, 18-25 nucleotide non-protein coding transcripts that play an important function in post-transcriptional regulation of gene expression during development. Specifically, we identified microRNA, miR-133b that is enriched in mDNs and functions within a regulatory feedback circuit with Pitx3. Here we propose to more broadly define the level of complexity of gene expression regulation by miRNA in mDNs, and to determine the function of these forms of regulation in vivo. Ultimately, such forms of regulation are likely to play a role in mDN-associated diseases, and furthermore manipulations of these mechanisms offer potential avenues for therapies. We wish to test two hypotheses: 1. miRNAs function in the regulation of mDNs, both within feedback circuits with mDN transcription factors and by the direct regulation of key mDN targets. 2. Such regulatory networks play functionally important roles in mDNs in vivo. PUBLIC HEALTH RELEVANCE A number of developmental and adult brain disorders are associated with midbrain dopaminergic neurons (mDNs), including Parkinson's disease (PD), schizophrenia, autism, dyskinesias, and drug addiction. Thus, the fundamental mechanisms that regulate the development and function of these cells are of great import. Here we propose to unravel the complex molecular regulatory signals that determine the development and function of midbrain dopamine neurons. We focus on the role of microRNAs, which are short RNA molecules that regulate the expression of key dopaminergic neuron genes. We initially use simplified model systems, including embryonic stem cell derived dopamine neurons and primary neuron cultures, which allow for a detailed molecular analysis. Ultimately, we extend these studies to confirming the role of regulatory molecular circuits in the intact behaving rodent.

描述（由申请人提供）：许多发育和成人脑部疾病与中脑多巴胺神经元（mDN）有关，包括帕金森病（PD）、精神分裂症、自闭症、运动障碍和药物成瘾。因此，调节这些细胞的发育和功能的基本机制非常重要。在之前的研究中，我们和其他人研究了基本的调节过程，例如在多巴胺神经元发育、功能和存活的背景下多巴胺生物合成途径中关键酶表达的调节。然而，从这些研究中可以清楚地看出，所有这些过程都具有高度的复杂性。例如，在简化的 ES 细胞培养模型中，至少 2 个转录因子 Nurr1 和 Pitx3 协同作用来控制晚期发育 mDN 标记的表达。最近，在初步数据和已发表的手稿中，我的实验室发现了额外一层复杂性的证据，涉及 mDN 发育和功能背景下 microRNA (miRNA) 的转录后调控。 miRNA 是进化上保守的 18-25 核苷酸非蛋白质编码转录物，在发育过程中基因表达的转录后调控中发挥重要作用。具体来说，我们鉴定了富含 mDN 的 microRNA miR-133b，并在 Pitx3 的调节反馈回路中发挥作用。在这里，我们建议更广泛地定义 mDN 中 miRNA 调节基因表达的复杂性水平，并确定这些调节形式在体内的功能。最终，这种形式的调节可能在 mDN 相关疾病中发挥作用，此外，对这些机制的操纵为治疗提供了潜在的途径。我们希望检验两个假设： 1. miRNA 在 mDN 的调节中发挥作用，既在 mDN 转录因子的反馈回路中发挥作用，又通过对关键 mDN 靶标的直接调节。 2. 这种调控网络在体内 mDN 中发挥着重要的功能作用。公共卫生相关性 许多发育和成人脑部疾病与中脑多巴胺能神经元 (mDN) 相关，包括帕金森病 (PD)、精神分裂症、自闭症、运动障碍和药物成瘾。因此，调节这些细胞的发育和功能的基本机制非常重要。在这里，我们建议解开决定中脑多巴胺神经元发育和功能的复杂分子调节信号。我们重点关注 microRNA 的作用，它们是调节关键多巴胺能神经元基因表达的短 RNA 分子。我们最初使用简化的模型系统，包括胚胎干细胞衍生的多巴胺神经元和原代神经元培养物，这可以进行详细的分子分析。最终，我们将这些研究扩展到确认调节分子电路在完整行为啮齿动物中的作用。