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

基本信息

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
项目类别：
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

项目摘要

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，以及吸毒过量的人数不断增加。然而，迄今为止，仍然没有足够的数据来检查分子。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一种多巴胺 (DA) 信号传导紊乱，但具有治疗作用。针对受体介导的 DA 神经传递的干预措施尚未完全有效因此，我们实验室的首要目标以及本应用的重点是我们的实验室研究 DA 参与介导成瘾表型的新颖、非常规行为。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，在我们实验室最近的努力中，通过转谷氨酰胺酶 2 (TGM2) 进行转酰胺基作用。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在大鼠中自我给药（SA），其中，gCaMKII：1)被发现仅在a下被强烈多巴胺酰化。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，其多巴胺酰化在急性期和长期禁欲，但不是对自然奖励的反应；3) 是参与的关键底物；介导从突触到大脑细胞核的长距离信号，最终促进 CREB 激活和因此，gCaMKII 上的多巴胺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。因此，我们认为 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。在目标 2 中，我们将探讨 gCaMKIIQ285dop 的吸毒与海洛因 SA 复发脆弱性。海洛因 SA 后对 CREB 信号/转录的影响，可能会导致复发脆弱性。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。