Contributions of aberrant synaptic protein monoaminylation to opiate use disorder

异常突触蛋白单胺化对阿片类药物使用障碍的影响

• 批准号: 10501338
• 负责人: DAVID M DIETZ
• 金额: $ 71.58万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-15 至 2027-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10501338
• 关键词: Abstinence; Acute; Affect; Amino Acids; Behavior; Behavioral; Binding; Brain; CREB1 gene; Calcium; Calmodulin; Cell Nucleus; Cells; ChIP-seq; Chemicals; Chronic; Corpus striatum structure; Coupled; Coupling; Cues; Data; Dependence; Deposition; Development; Discipline; Disease; Dopamine; Dopamine D2 Receptor; Drug Addiction; Enzymes; Event; Female; Future; Genetic Transcription; Glutamine; Goals; Health; Heroin; Histone H3; In Vitro; Individual; Laboratories; Learning; Link; Mass Spectrum Analysis; Measures; Mediating; Memory; Molecular; Molecular Abnormality; Neurobiology; Neuronal Plasticity; Neurons; Neurophysiology - biologic function; Neurosciences; Nuclear; Nuclear Translocation; Nucleus Accumbens; Open Reading Frames; Opiate Addiction; Opioid; Overdose; Pharmaceutical Preparations; Phenotype; Phosphorylation; Phosphotransferases; Play; Population; Post-Translational Protein Processing; Predisposition; Protein Isoforms; Proteins; Psychological reinforcement; Public Health; Rattus; Recombinants; Recurrent disease; Regulation; Relapse; Rewards; Rodent Model; Role; Self Administration; Serine; Signal Transduction; Site; Source; Sucrose; Synapses; Synaptic plasticity; Therapeutic Intervention; Threonine; Transgenic Organisms; Up-Regulation; Viral; Viral Genes; Virus; Volition; Western Blotting; addiction; behavioral pharmacology; behavioral plasticity; brain reward regions; calmodulin-dependent protein kinase II; cell type; drug abstinence; drug withdrawal; effective intervention; effective therapy; efficacious treatment; epigenomics; gene therapy; genetic manipulation; in vivo; male; mutant; neurobiological mechanism; neurotransmission; novel; opioid abuse; opioid use; opioid use disorder; prevent; psychostimulant; receptor; response; therapeutically effective; transcription factor; transcriptome sequencing; transglutaminase 2

PROJECT SUMMARY Opiate use, dependence and addiction represent enduring public health issues, resulting in substantial financial and societal health burdens, as well as increasing numbers of overdoses. Drug addiction is characterized as a chronic, relapsing disease. However, to date, there remains insufficient data examining the molecular mechanisms underlying persistent opiate-induced neurobiological changes, which has led to a scarcity of effective therapies and interventions to treat and prevent relapse. Drug addiction has long been thought of as a disorder of dopamine (DA) signaling. However, therapeutic interventions targeting receptor mediated DA neurotransmission have not yet resulted in fully efficacious treatments. Therefore, an overarching goal of our laboratories – and the focus of this application – has been to investigate novel, non-canonical actions of DA involved in mediating addiction phenotypes. Our laboratory recently identified a novel signaling moiety for DA in brain, termed dopaminylation (dop), whereby DA acts as a donor source for the establishment of post-translational modifications (PTM) on substrate proteins (e.g., histone H3) via transamidation by the Transglutaminase 2 (TGM2) enzyme. In more recent efforts by our lab to unbiasedly identify additional substrates of these PTMs (focused now on synaptic proteins in nucleus accumbens/NAc, a key brain reward region), we developed a novel chemical tagging approach that, when coupled to mass spectrometry, allowed for the discovery of hundreds of dopaminylated proteins in brain, both in the context of normal neural function and in response to aberrant dopamine signaling following chronic heroin self-administration (SA) in rats. Among them, gCaMKII: 1) was found to be robustly dopaminylated at only a single amino acid residue located within its autoinhibitory helix [glutamine (Q)285], a site that exists only two amino acids away from a critical threonine (T) residue (287), which when phosphorylated directs Calmodulin (CaM) sequestration; 2) is upregulated in its dopaminylation following heroin SA, both during acute and prolonged abstinence, but not in response to natural rewards; and 3) represents a critical substrate involved in mediating long range signals from the synapse to nucleus in brain, ultimately promoting CREB activation and neuronal plasticity. Thus, this dopaminylation event on gCaMKII may represent a critical convergent mechanism linking altered dopaminergic signaling in response to heroin to CREB mediated transcriptional abnormalities. As such, we hypothesize that gCaMKIIQ285dop may play a direct role in mediating heroin relapse via aberrant modulation of CREB signaling in NAc. In Aim 1, we will fully characterize gCaMKIIQ285dop’s temporal effects on drug taking vs. relapse vulnerability in the context of heroin SA. In Aim 2, we will explore gCaMKIIQ285dop’s effects on CREB signaling/transcription following heroin SA, events that may precipitate relapse vulnerability. In Aim 3, we will investigate roles for gCaMKIIQ285dop mediated CREB signaling/transcription in D1 vs. D2 dopamine receptor-expressing MSNs during abstinence from heroin SA in the regulation of relapse behaviors.

项目概要 阿片剂的使用、依赖和成瘾是持久的公共卫生问题，导致大量的经济损失 和社会健康负担，以及药物过量数量的增加。吸毒成瘾的特征是 慢性、复发性疾病。然而，迄今为止，仍然没有足够的数据来检验分子 持续阿片类药物引起的神经生物学变化的潜在机制，这导致了阿片类药物的稀缺 治疗和预防复发的有效疗法和干预措施。 长期以来，药物成瘾一直被认为是多巴胺 (DA) 信号传导障碍。然而，治疗 针对受体介导的 DA 神经传递的干预措施尚未完全有效 治疗。因此，我们实验室的总体目标以及本应用的重点是 研究 DA 参与介导成瘾表型的新颖、非常规行为。我们的实验室 最近在大脑中发现了一种新的 DA 信号传导部分，称为多巴胺化 (dop)，其中 DA 充当 用于在底物蛋白上建立翻译后修饰 (PTM) 的供体来源（例如， 组蛋白 H3) 通过转谷氨酰胺酶 2 (TGM2) 进行转酰胺基作用。在我们实验室最近的努力中 公正地识别这些 PTM 的其他底物（现在重点关注细胞核中的突触蛋白 伏隔核/NAc，一个关键的大脑奖励区域），我们开发了一种新颖的化学标记方法，当 与质谱联用，允许在大脑中发现数百种多巴胺酰化蛋白质，无论是在 正常神经功能的背景以及对慢性海洛因后异常多巴胺信号的反应 大鼠自我给药（SA）。其中，gCaMKII：1)被发现仅在 位于其自抑制螺旋 [谷氨酰胺 (Q)285] 内的单个氨基酸残基，该位点仅存在两个 远离关键苏氨酸 (T) 残基 (287) 的氨基酸，磷酸化后可指导钙调蛋白 (CaM) 隔离； 2) 在海洛因 SA 后，其多巴胺酰化水平上调，无论是在急性期还是 长期禁欲，但不是对自然奖励的反应； 3）代表参与的关键底物 介导从突触到大脑细胞核的长距离信号，最终促进 CREB ​​激活和 神经元的可塑性。因此，gCaMKII 上的多巴胺酰化事件可能代表了一种关键的收敛机制 将响应海洛因的多巴胺能信号改变与 CREB ​​介导的转录异常联系起来。作为 因此，我们假设 gCaMKIIQ285dop 可能通过异常在介导海洛因复吸中发挥直接作用 NAc 中 CREB ​​信号传导的调节。在目标 1 中，我们将全面表征 gCaMKIIQ285dop 的时间效应 关于海洛因 SA 背景下的吸毒与复发脆弱性。在目标 2 中，我们将探索 gCaMKIIQ285dop 的 海洛因 SA 后对 CREB ​​信号/转录的影响，可能会导致复吸的事件。在 目标 3，我们将研究 gCaMKIIQ285dop 介导的 CREB ​​信号/转录在 D1 与 D2 中的作用 戒除海洛因 SA 期间表达多巴胺受体的 MSN 对复发行为的调节。