Autophagic regulation of cocaine abuse

可卡因滥用的自噬调节

• 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
• 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

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.

项目摘要/摘要 该多PI R01应用程序的目的是确定自噬机械如何调节 滥用可卡因的发展。可卡因是最广泛滥用的药物之一，并产生多种 行为包括奖励，渴望和复发。尽管许多药理目标和行为 已经探讨了干预措施，没有FDA批准的药物来减少可卡因的使用或治疗 可卡因成瘾者复发。自噬是靶向cargos的关键溶酶体降解途径 选择性或非选择性降解，在时间和空间上受到30以上的控制 自噬基因。自噬是在基础水平的组成型发生的，并且可以通过压力进一步诱导。我们 最近发现，与自噬相关的蛋白BecN2/Beclin 2，与 自噬诱导III类磷脂酰肌醇3-激酶VPS34是可卡因采集的新调节剂 通过多巴胺（DA）D2受体（D2RS）奖励行为。 BECN2的敲除（但不是其同源性BECN1） 全球或专门在DA神经元中保护小鼠免受可卡因诱导的运动刺激的影响 放置偏好和静脉自我管理。 UPLC/质谱分析表明可卡因 - DA的诱导积累，而不是其他神经递质的累积，这会因BECN2耗竭而减弱。此外， BECN2上游的药理学抑制自噬激酶，包括ULK1和VPS34，模仿 BECN2耗竭/突变对可卡因诱导的奖励行为和DA积累的影响，这表明 在可卡因反应调节中存在ULK1-VPS34-BECN2轴的存在。此外，生化 分析表明，D2R是通过与BECN2相关蛋白质GASP1结合的BECN2的降解靶标。 遗传抑制BECN2或对ULK1的药理抑制类似地增加了纹状体突触前D2RS。 基于这些初步数据，我们建议研究ULK1-VPS34-BECN2的假设 自噬轴通过选择性调节D2R自身受体溶血体来控制可卡因滥用的脆弱性 DA神经元的贩运和降解。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神经元中的功能如何 调节可卡因摄取，剂量反应，恢复可卡因的剂量和D2R分解代谢的获取 （目标1）？在调节这些可卡因奖励的DA神经元中，是否有ULK1-VPS34-BECN2自噬途径 行为，如果是这样，它如何工作（目标2）？我们预计，随着这两个根本相关的目标， 我们的研究将确定以BECN2为中心自噬的功能和机制在D2调节中 受体代谢，daergic功能和与可卡因相关的奖励行为。对此有更好的理解 新颖的分子机制可能为开发可卡因滥用的治疗方法提供了新的选择 药物滥用类型。