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

• 批准号: 10512656
• 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
• 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/10512656
• 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; Lead; Life; Link; Metabolic; Mus; Neurobiology; Neuronal Plasticity; Neurons; Opiate Addiction; Opioid; Output; Parvalbumins; Play; 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; preclinical study; repaired; response; reward circuitry; transcriptome; translatome

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，尽管治疗了当前的治疗，但复发率很高。临床 临床前研究发现，阿片类药物诱导的基因表达慢性变化是神经塑性的 大脑奖励电路内的变化，包括前额叶皮层（PFC），导致终身 复发的倾向。最近的研究强调，表观遗传变化策划了阿片类药物滥用引起的 转录可塑性。但是，表观基因组变化的原因和随之而来的转录组 改变仍然不清楚。 DNA损伤和维修过程在与适当的染色质完整性中起重要作用 细胞中的表观遗传学特征和基因表达。有丝分裂后神经元特别容易受到DNA的影响 由于其高代谢和转录输出而损坏。 DNA断裂触发DNA损伤修复 反应，涉及大量染色质重塑以融合DNA修复机械。因此，DNA 断裂和维修可以逐渐改变染色质结构以改变神经元中的基因表达模式，并改变 最终，影响行为。 与将DNA损伤和阿片类药物成瘾联系起来的研究一致，我们的初步数据表明DNA 来自阿片类药物使用障碍的个体的死后PFC组织中的断裂增加了 那种自我管理的海洛因（阿片类药物成瘾的有效动物模型）。另外，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， 在PFC中引入DNA断裂的实验性，在小鼠中增强了海洛因的寻求行为。在一起， 这些初步数据提供了证据表明DNA断裂和修复过程与阿片类药物成瘾有关。我们的 该提案中的目标是识别和表征DNA断裂和维修在阿片类药物成瘾中的精确作用。 使用基因组，表观基因组和翻译方法的独特组合，结合亲和力 纯化，我们将充分表征海洛因引起的神经元特异性（PFC白蛋白中间神经元和 DNA断裂和修复位点（即断裂）和相关的变化的锥体神经元）的改变 在染色质的可及性（即表观基因组）和基因表达（即翻译组）中。另外，使用 CRISPR/CAS9基因组编辑方法，我们将确定海洛因诱导的DNA断裂变化和 维修地点直接与因果关系与海洛因诱导的行为可塑性有关。 该提案将为各种未来研究的努力提供稳固的基础，以解决 阿片类药物诱导的长期转录疾病的基础的遗传和表观遗传机制和 最终有助于阿片类药物成瘾。