The role of DNA breaks and repairs in opioid addiction

DNA 断裂和修复在阿片类药物成瘾中的作用

基本信息

批准号：
10704731
负责人：
Zijun Wang
金额：
$ 45.9万
依托单位：
UNIVERSITY OF KANSAS LAWRENCE
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-15 至 2027-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10704731
关键词：
Address Affect Affinity Chromatography Animal Model Autopsy Behavior Behavioral Brain CRISPR/Cas technology Cells Chromatin Chromatin Structure Chronic Clinical Research Coupled DNA DNA Damage DNA Repair Data Disease Drug Addiction Epigenetic Process Foundations Gene Expression Gene Expression Profile Genetic Genetic Transcription Genomics Goals Heroin Individual Interneurons Link Metabolic Mus Neurobiology Neuronal Plasticity Neurons Opiate Addiction Opioid Output Parvalbumins Play Predisposition Prefrontal Cortex Process Relapse Role Self Administration Site Solid Tissues Transcriptional Regulation behavioral plasticity chromatin remodeling epigenome epigenomics genome editing hippocampal pyramidal neuron novel opioid abuse opioid use disorder postmitotic preclinical study repaired response reward circuitry transcriptome translatome

项目摘要

Abstract Opioid use disorder is a debilitating disease with a high rate of relapse despite current treatment efforts. Clinical and preclinical studies have found that opioid-induced chronic changes in gene expression underlie neuroplastic alterations within the brain reward circuitry, including in the prefrontal cortex (PFC), which lead to the life-long propensity for relapse. Recent studies highlighted that epigenetic changes orchestrate the opioid abuse-induced transcriptional plasticity. However, the causes for the epigenome changes and consequent transcriptome alterations remain unclear. DNA damage and repair processes play important roles in chromatin integrity relating to proper epigenetic signature and gene expression in cells. Postmitotic neurons are particularly susceptible to DNA damage because of their high metabolic and transcriptional output. DNA breaks trigger the DNA damage repair response, which involves extensive chromatin remodeling to incorporate DNA repair machinery. Therefore, DNA breaks and repairs can progressively alter chromatin structure to alter gene expression patterns in neurons and, ultimately, affect behaviors. Consistent with studies linking DNA damage and opioid addiction, our preliminary data suggest that DNA breaks are increased in the postmortem PFC tissues from individuals with opioid use disorder and from mice that self-administered heroin (a valid animal model for opioid addiction). Additionally, the increase in DNA breaks was accompanied by altered expression of genes that are involved in DNA damage repair. More importantly, experimentally introducing DNA breaks in the PFC potentiated heroin-seeking behavior in mice. Taken together, these preliminary data provide evidence that DNA break and repair processes are linked to opioid addiction. Our goal in this proposal is to identify and characterize the precise role of DNA breaks and repairs in opioid addiction. Using a unique combination of genomic, epigenomic, and translatomic approaches coupled with affinity purification, we will fully characterize heroin-induced neuron-specific (PFC parvalbumin interneurons and pyramidal neurons) alterations in DNA break and repair sites (i.e., the breakome) and the associated changes in chromatin accessibility (i.e., the epigenome) and gene expression (i.e., the translatome). Additionally, using CRISPR/Cas9 genome editing approaches, we will identify whether heroin-induced changes in DNA break and repair sites are directly and causally linked to heroin-induced behavioral plasticity. This proposal will provide a solid foundation for a wide range of future research endeavors addressing the genetic and epigenetic mechanisms that underlie opioid-induced long-term transcriptional maladaptation and ultimately contribute to opioid addiction.

抽象的阿片类药物使用障碍是一种使人衰弱的疾病，尽管目前采取了治疗措施，但复发率很高。临床临床前研究发现阿片类药物引起的基因表达的慢性变化是神经可塑性的基础大脑奖赏回路的改变，包括前额皮质（PFC），这会导致终生复发倾向。最近的研究强调，表观遗传变化协调了阿片类药物滥用引起的转录可塑性。然而，表观基因组变化和随之而来的转录组变化的原因变化仍不清楚。 DNA 损伤和修复过程在染色质完整性中发挥着重要作用，与正确的染色质完整性相关。细胞中的表观遗传特征和基因表达。有丝分裂后神经元对 DNA 特别敏感由于其高代谢和转录输出而造成损害。 DNA断裂触发DNA损伤修复反应，涉及广泛的染色质重塑以纳入 DNA 修复机制。因此，DNA 断裂和修复可以逐渐改变染色质结构，从而改变神经元中的基因表达模式，最终影响行为。与 DNA 损伤和阿片类药物成瘾相关的研究一致，我们的初步数据表明 DNA 阿片类药物使用障碍个体和小鼠死后 PFC 组织中的断裂增加自我注射海洛因（阿片类药物成瘾的有效动物模型）。此外，DNA 断裂的增加伴随着参与 DNA 损伤修复的基因表达的改变。更重要的是，实验性地在 PFC 中引入 DNA 断裂增强了小鼠的海洛因寻求行为。综合起来，这些初步数据证明 DNA 断裂和修复过程与阿片类药物成瘾有关。我们的该提案的目标是确定和描述 DNA 断裂和修复在阿片类药物成瘾中的确切作用。使用基因组学、表观基因组学和翻译原子学方法的独特组合以及亲和力纯化后，我们将充分表征海洛因诱导的神经元特异性（PFC 小白蛋白中间神经元和锥体神经元）DNA断裂和修复位点（即断裂组）的改变以及相关的变化染色质可及性（即表观基因组）和基因表达（即翻译组）。此外，使用 CRISPR/Cas9基因组编辑方法，我们将确定海洛因是否引起DNA断裂和变化修复位点与海洛因引起的行为可塑性有直接的因果关系。该提案将为未来解决这一问题的广泛研究工作奠定坚实的基础。阿片类药物引起的长期转录适应不良的遗传和表观遗传机制最终导致阿片类药物成瘾。