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：
10681305
负责人：
DAVID M DIETZ
金额：
$ 69.25万
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/10681305
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 Rat Transgene Rattus Recombinants Recurrent disease Regulation Relapse Rewards Rodent Model Role Self Administration Serine Signal Transduction Site Source Sucrose Synapses Synaptic plasticity Therapeutic Intervention Threonine Up-Regulation Viral Viral Genes Virus Western Blotting addiction amidation 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 induced pluripotent stem cell inducible Cre male mutant neurobiological mechanism neurotransmission novel opioid abuse opioid use opioid use disorder prolonged abstinence psychostimulant receptor relapse prevention 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.

项目摘要 opate使用，依赖和成瘾代表持久的公共卫生问题，导致大量财务和社会健康伯恩斯（Burnens）以及越来越多的过量服用。吸毒成瘾的特征是慢性接力疾病。但是，迄今为止，数据仍不足以检查分子持续操作引起的神经生物学变化的机制，这导致了稀缺性有效的疗法和干预措施以治疗和防止退休。长期以来，吸毒成瘾被认为是多巴胺（DA）信号传导的疾病。但是，治疗靶向受体介导的DA神经传递的干预措施尚未达到完全有效的治疗。因此，我们实验室的总体目标以及本应用的重点是研究与中介成瘾表型有关的新型DA的非传统作用。我们的实验室最近确定了大脑中DA的新型信号传导部分，称为多巴胺基化（DOP），从而充当DA 建立底物蛋白上翻译后修饰（PTM）的供体来源（例如 HISSTONE H3）通过转谷氨酰胺酶2（TGM2）酶通过透射增强。在我们实验室的最新努力中公正地识别这些PTM的其他底物（现在集中在核中的合成蛋白上 Accumbens/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，当结合到质谱法，允许在大脑中发现数百种多巴胺基化蛋白正常神经元功能和慢性海洛因后异常多巴胺信号传导的背景大鼠的自我管理（SA）。其中，发现GCAMKII：1）仅在位于其自身抑制螺旋内的单氨基酸住宅[谷氨酰胺（Q）285]，该位点仅存在两个氨基酸远离关键苏氨酸（T）居住（287），当磷酸化时引导钙调蛋白（CAM）隔离； 2）在急性和节制长期，但没有回应自然的奖励； 3）代表涉及的关键基材介导从突触到大脑中核的远程信号，最终促进CREB激活和神经元可塑性。那就是GCAMKII上的这种多巴胺基化事件可能代表关键的收敛机制将改变海洛因的多巴胺能信号与CREB介导的转录异常联系起来。作为这样，我们假设GCAMKIIQ285DOP可能在通过异常中介导海洛因继电器中发挥直接作用 NAC中CREB信号的调制。在AIM 1中，我们将充分表征GCAMKIIQ285DOP的临时效果关于在海洛因SA的情况下吸毒与救济脆弱性。在AIM 2中，我们将探索GCAMKIIQ285DOP 对海洛因SA后CREB信号传导/转录的影响，可能会导致继电器脆弱性的事件。在 AIM 3，我们将研究GCAMKIIQ285DOP介导的CREB信号/转录D1与D2的角色在对继电器行为的调节中，海洛因SA戒酒期间表达多巴胺受体的MSN。