Neuron subtype specific role of DNA methylcytosine dioxygenase TET1 in cocaine addiction

DNA甲基胞嘧啶双加氧酶TET1在可卡因成瘾中的神经元亚型特异性作用

基本信息

批准号：
10177985
负责人：
Jian Feng
金额：
$ 34.09万
依托单位：
FLORIDA STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-09-15 至 2023-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10177985
关键词：
Address Affect Behavior Behavioral Brain CRISPR/Cas technology Cells Chronic Cocaine Cocaine Dependence Coupled Cytosine DNA DNA Methylation DNA Modification Process DNA Sequence DNA methylation profiling Dinucleoside Phosphates Dioxygenases Dopamine Dopamine D2 Receptor Drug Addiction Economic Burden Enzymes Epigenetic Process Family Family member Future Gene Expression Gene Silencing Genes Genetic Transcription Genomic DNA Goals Health Human Interphase Cell Lead Literature Mediating Molecular Morphology Mus Neuronal Plasticity Neurons Neurophysiology - biologic function Nucleus Accumbens Oxides Pathway interactions Pattern Play Population Projection Positioning Attribute Process Protein translocation Reporting Resolution Rewards Role Site Stimulus Testing Therapeutic Time Validation Ventral Striatum Work addiction base brain reward regions brain tissue cell type conditional knockout demethylation drug reward epigenome editing member methyl group novel promoter response reward circuitry social whole genome

项目摘要

Neural function requires accurate control of gene transcription in response to environmental stimuli. Epigenetics plays a crucial role in this process by modulating gene expression without changing underlying DNA sequences. DNA methylation is a major epigenetic mechanism, wherein a methyl group is covalently coupled to the C5 position of cytosine, predominantly at 5'-CpG-3' dinucleotide sites. Though still few in number, accumulating evidence demonstrates that DNA methylation may play important roles in drug addiction. However, to firmly establish DNA methylation's role in drug addiction, the question of whether and how DNA methylation changes take place and function in neurons must be addressed. This is particularly true for those defined neuronal subtypes selectively engaged in drug addiction. To address this question, we herein propose to study methylcytosine dioxygenase TET1, a newly identified DNA demethylation enzyme, and its mediated DNA methylation turnover in the two major neuron types (D1- and D2- medium spiny neurons (MSNs)) in nucleus accumbens in cocaine addiction. In the past, we have found TET1, but not TET2 or TET3 (the other two members of the dioxygenase family), is selectively regulated by cocaine in the nucleus accumbens. We have also shown TET1 mediates drug reward behaviors. Here, we propose TET1 carries distinct functions in D1- and D2-MSNs, which usually play opposite roles in addiction reward circuitry. We will also elucidate TET1-mediated neuron subtype-specific DNA methylation changes in D1- and D2-MSNs. Lastly, we will probe the long-standing question of the causal functional role of DNA methylation in drug addiction-relevant behavior through a proven CRISPR-Cas9 based epigenome editing approach to modify DNA methylation at selective loci. Upon completion, our study will not only advance our scientific understanding of DNA epigenetic underpinnings of drug addiction in a neuron subtype-specific manner, it will also provide a path to manipulate behaviors associated with drug addiction through cell type- specific precision epigenome editing, which has obvious potential utility for future therapeutic applications.

神经功能需要对环境刺激的响应进行准确控制基因转录。表观遗传学在此过程中通过调节基因表达而不改变潜在的基因表达起着至关重要的作用 DNA序列。 DNA甲基化是一种主要的表观遗传机制，其中甲基是共价的耦合到胞嘧啶的C5位置，主要是在5'-CPG-3'二核苷酸位点。虽然很少数量，积累的证据表明DNA甲基化可能在药物中起重要作用瘾。但是，要牢固确定DNA甲基化在药物成瘾中的作用必须解决如何在神经元中进行DNA甲基化变化。尤其是对于那些定义的神经元亚型选择性地参与药物成瘾的人是正确的。为了解决这个问题，我们在这里建议研究甲基胞嘧啶二氧酶TET1，这是一种新鉴定的DNA脱甲基化酶及其介导的两种主要神经元类型的DNA甲基化周转率（D1-和D2-培养基可卡因成瘾中伏核核中的棘神经元（MSNS））。过去，我们找到了Tet1，但是不是TET2或TET3（二氧酶家族的其他两个成员），由可卡因选择性地调节伏隔核。我们还显示了TET1介导了药物奖励行为。在这里，我们建议 TET1在D1和D2-MSN中具有不同的功能，通常在成瘾中起作用的角色相反电路。我们还将阐明D1-的TET1介导的神经元亚型特异性DNA甲基化变化和D2-MSN。最后，我们将探讨DNA因果功能作用的长期问题通过验证的基于CRISPR-CAS9的表观基因组编辑，与药物成瘾相关的行为中的甲基化在选择性位点修饰DNA甲基化的方法。完成后，我们的研究不仅会推进我们的对神经元亚型特异性药物成瘾的DNA表观遗传基础的科学理解方式，它还将提供通过细胞类型来操纵与药物成瘾相关行为的途径 - 特定的精度表观基因组编辑，具有明显的对未来治疗应用的潜在效用。