Autophagic regulation of cocaine abuse

可卡因滥用的自噬调节

基本信息

批准号：
10682457
负责人：
Congcong He
金额：
$ 53.04万
依托单位：
NORTHWESTERN UNIVERSITY AT CHICAGO
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/10682457
关键词：
Attenuated Autophagocytosis Autoreceptors Behavior Behavior Therapy Behavioral Binding Biochemical Catabolism Cell Surface Receptors Cellular biology Cocaine Cocaine Abuse Cocaine Dependence Complex Corpus striatum structure Data Degradation Pathway Development Dopamine Dopamine D2 Receptor Dose Drug abuse FDA approved Future GPRASP1 gene Genes Genetic Homologous Gene Image Intravenous Knock-out Knockout Mice Link Locomotion Mass Spectrum Analysis Metabolism Molecular Mus Mutate Mutation Neurons Neurotransmitters Output Pathogenicity Pathway interactions Pharmaceutical Preparations Phenocopy Proteins Regulation Relapse Research Rewards Role Self Administration Signal Transduction Stress System Testing Time Vps34 Phosphatidylinositol 3 Kinase Wild Type Mouse Work cocaine reward cocaine seeking cocaine use conditioned place preference craving dopaminergic neuron drug of abuse extracellular inhibition of autophagy novel overdose death pharmacologic presynaptic prevent receptor response reuptake targeted treatment tool trafficking upstream kinase

项目摘要

Project Summary/Abstract The objective of this multi-PI R01 application is to determine how the autophagy machinery regulates the development of cocaine abuse. Cocaine is one of the most widely abused drugs, and produces a variety of behaviors including reward, craving, and relapse. Although many pharmacological targets and behavioral interventions have been explored, there are no FDA-approved medications for reducing cocaine use or treating relapse in cocaine addicts. Autophagy is a key lysosomal degradation pathway that targets cargos for degradation either selectively or non-selectively, and is temporally and spatially controlled by more than 30 autophagy genes. Autophagy occurs constitutively at a basal level, and can be further induced by stress. We recently discovered that an autophagy-related protein Becn2/Beclin 2, which forms a complex with the autophagy-inducing class III phosphatidylinositol 3-kinase Vps34, is a novel regulator of acquisition of cocaine reward behaviors via dopamine (DA) D2 receptors (D2Rs). Knockout of Becn2 (but not its homolog Becn1) globally or specifically in DA neurons protects mice from cocaine-induced locomotor stimulation, conditioned place preference, and intravenous self-administration. UPLC/mass spectrometry profiling indicates that cocaine- induced accumulation of DA, but not other neurotransmitters, is attenuated by Becn2 depletion. In addition, pharmacologically inhibiting autophagy kinases upstream of Becn2, including ULK1 and Vps34, mimics the effects of Becn2 depletion/mutation on cocaine-induced reward behaviors and DA accumulation, suggesting the existence of a ULK1-Vps34-Becn2 axis in the regulation of cocaine responses. Furthermore, biochemical analyses reveal that D2R is a degradation target of Becn2 via binding to a Becn2-associated protein GASP1. Genetic inhibition of Becn2 or pharmacological inhibition of ULK1 similarly increases striatal presynaptic D2Rs. Based on these preliminary data, we propose to investigate the hypothesis that the ULK1-Vps34-Becn2 autophagy axis controls vulnerability to cocaine abuse by selectively regulating D2R autoreceptor endolysosomal trafficking and degradation in DA neurons. Using a combination of genetic, imaging, biochemical, cellular and behavioral approaches, we aim to answer the following questions: How does Becn2 function in DA neurons to regulate acquisition of cocaine-taking, dose response, reinstatement of cocaine-seeking, and D2R catabolism (Aim 1)? Is there a ULK1-Vps34-Becn2 autophagy pathway in DA neurons regulating these cocaine reward behaviors, and if so, how does it work (Aim 2)? We anticipate that with these two fundamentally related aims, our study will establish the function and mechanism of a Becn2-centered autophagy axis in the regulation of D2 receptor metabolism, DAergic function, and cocaine-related reward behaviors. A better understanding of this novel molecular mechanism may provide new options for developing treatments for cocaine abuse and additional types of drug abuse.

项目概要/摘要该多 PI R01 应用的目的是确定自噬机制如何调节可卡因滥用的发展。可卡因是最广泛滥用的药物之一，产生多种行为包括奖励、渴望和复发。尽管许多药理学目标和行为已经探索了干预措施，但 FDA 还没有批准药物来减少可卡因使用或治疗可卡因成瘾者复吸。自噬是一种关键的溶酶体降解途径，其目标是货物选择性或非选择性降解，并且在时间和空间上受 30 多个因素控制自噬基因。自噬在基础水平上持续发生，并且可以通过应激进一步诱导。我们最近发现一种与自噬相关的蛋白Becn2/Beclin 2，它与自噬形成复合物自噬诱导 III 类磷脂酰肌醇 3-激酶 Vps34 是一种新型可卡因获取调节剂通过多巴胺 (DA) D2 受体 (D2R) 奖励行为。 Becn2 的敲除（但不是其同源 Becn1）全局或特别是在 DA 神经元中保护小鼠免受可卡因诱导的运动刺激（条件反射）地点偏好和静脉自我给药。 UPLC/质谱分析表明可卡因- Becn2 耗尽会减弱诱导的 DA 积累，但不会减弱其他神经递质的积累。此外，药理学上抑制 Becn2 上游的自噬激酶，包括 ULK1 和 Vps34，模仿 Becn2 缺失/突变对可卡因诱导的奖赏行为和 DA 积累的影响，表明 ULK1-Vps34-Becn2 轴在可卡因反应调节中的存在。此外，生化分析表明，D2R 通过与 Becn2 相关蛋白 GASP1 结合而成为 Becn2 的降解靶标。 Becn2 的遗传抑制或 ULK1 的药理学抑制同样会增加纹状体突触前 D2R。基于这些初步数据，我们建议研究以下假设：ULK1-Vps34-Becn2 自噬轴通过选择性调节 D2R 自受体内溶酶体来控制可卡因滥用的脆弱性 DA 神经元的运输和降解。结合遗传、成像、生化、细胞和行为方法，我们旨在回答以下问题：Becn2 在 DA 神经元中如何发挥作用调节可卡因的获取、剂量反应、可卡因寻求的恢复和 D2R 分解代谢（目标1）？ DA 神经元中是否存在 ULK1-Vps34-Becn2 自噬通路来调节这些可卡因奖赏行为，如果是，它是如何运作的（目标 2）？我们预计，通过这两个根本相关的目标，我们的研究将建立以Becn2为中心的自噬轴在D2调节中的功能和机制受体代谢、DAergic 功能和可卡因相关的奖励行为。对此有更好的理解新的分子机制可能为开发可卡因滥用和其他治疗方法提供新的选择药物滥用的类型。