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Identification of CaMKII as a novel substrate of dopaminylation following heroin self administration

CaMKII 鉴定为海洛因自我给药后多巴胺酰化的新型底物

基本信息

批准号：
10555211
负责人：
Andrew F Stewart
金额：
$ 4.61万
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：
2022-01-19 至 2024-01-18
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10555211
关键词：
Abstinence Acute Affect Alanine Amino Acids Back Behavior Behavior Control Binding Brain Calcium Calmodulin Cell Culture Techniques Cell Nucleus Cells Cessation of life ChIP-seq Chemicals Chronic Co-Immunoprecipitations Complex Corpus striatum structure Coupled Coupling Cues Custom Cyclic AMP Response Element Data Disease Dopamine Economic Burden Enzymes Epigenetic Process Event Functional disorder Gene Expression Profile Genetic Transcription Glutamine Heroin Histone H3 In Vitro Infiltration Intervention Laboratories Learning Link Long-Term Potentiation Mass Spectrum Analysis Mediating Memory Modeling Mutate Neurons Neurophysiology - biologic function Nuclear Nuclear Translocation Nucleus Accumbens Opioid Pharmaceutical Preparations Phosphorylation Phosphotransferases Play Post-Translational Protein Processing Proteins Public Health Rattus Recombinants Regulation Relapse Research Rewards Role Saline Self Administration Signal Transduction Site Source Substance Use Disorder Substance abuse problem Sucrose Synapses Synaptic plasticity Testing Threonine Tissues United States Validation Ventral Tegmental Area Viral Virus Western Blotting adeno-associated viral vector amidation behavioral outcome calmodulin-dependent protein kinase II drug of abuse drug seeking behavior efficacious treatment experimental study heroin use in vivo knock-down mesolimbic system monoamine mutant neurobiological mechanism new therapeutic target novel postsynaptic prolonged abstinence receptor response socioeconomics transcription factor transcriptome sequencing transglutaminase 2

项目摘要

Heroin use disorder (HUD) represents an enduring public health issue resulting in significant socioeconomic burdens to the United States, with domestic opiate-related deaths quadrupling from 1999 to 2017. Despite this the neurobiological mechanisms underlying HUD remain poorly understood. All drugs of abuse modulate dopaminergic signaling and have long been thought of as disorders of dopamine (DA) signaling. However, pharmacotherapeutic interventions targeting receptor mediated DA-signaling have not resulted in efficacious treatments. Our laboratory recently identified a novel signaling moiety for DA, termed dopaminylation, whereby DA itself acts as a post-translational modification (PTM) on substrate proteins via transamidation by the Transglutaminase 2 (TGM2) enzyme. I sought, then, to unbiasedly identify additional synaptic substrates of dopaminylation in vivo, utilizing a novel chemical tagging approach coupled to mass spectrometry. I identified 164 novel putative synaptic substrates of dopaminylation in Nucleus Accumbens (NAc), both in the context of normal neural function and in response to abstinence from chronic heroin self-administration. Following validation of a number of putative substrates, I turned my focus to gCaMKII as: 1) it is highly abundant 2) it is dopaminylated at a single amino acid residue, located within it’s autoinhibitory helix (glutamine [Q]285), a site that exists only two amino acids away from the critical threonine (T) residue 287. T287 is phosphorylated to direct Calmodulin (CaM) sequestration and subsequent nuclear translocation – thus, this site, represents an exciting ‘test’ case for establishing the importance of this signaling moiety in post synaptic plasticity 3) it is upregulated in its dopaminylation state following abstinence from heroin SA, an effect that persists to AD14 and 4)represents a critical substrate involved in mediating long range signals from the synapse to the nucleus in brain, ultimately promoting CREB activation. gCaMKII is a synaptic protein demonstrated to be necessary for learning/memory, late-long-term-potentiation (LTP) and excitation transcription coupling. CREB is an important transcription factor in brain implicated in all types of SUD, where dynamic alterations in activation have been observed to affect drug-related behaviors. I have additionally demonstrated that monoamine molecules can infiltrate striatal and cortical neurons in vitro, as well as that treatment of these neurons with DA induces gCaMKII dopaminylation. I also observed the dynamic regulation of gCaMKII/CaM localization and subsequent downstream CREB signaling in response to DA treatment in primary cortical-striatal neuronal culture. I therefore hypothesize that gCaMKIIQ285dop may represent a novel dopaminergic signaling moiety in brain, and may play a direct role in mediating heroin relapse behaviors via aberrant modulation of CREB signaling in NAc.

海洛因使用障碍（HUD）是一个持久的公共卫生问题，导致重大的社会经济