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.

项目总结/摘要 这种多PI R 01应用的目的是确定自噬机制如何调节细胞的增殖。 可卡因滥用的发展。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。尽管许多药理学靶点和行为学靶点 虽然已经探索了干预措施，但没有FDA批准的药物用于减少可卡因的使用或治疗 可卡因成瘾者的复吸自噬是一种关键的溶酶体降解途径，靶向货物， 选择性或非选择性地降解，并且在时间和空间上由30多个 自噬基因自噬在基础水平上组成性发生，并且可以通过应激进一步诱导。我们 最近发现一种自噬相关蛋白Becn 2/Beclin 2，它与 自噬诱导III类磷脂酰肌醇3-激酶Vps 34，是一种新的可卡因获得调节剂 通过多巴胺（DA）D2受体（D2 Rs）的奖励行为。敲除Becn 2（但不是其同系物Becn 1） 整体或特异性地在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， 位置偏好和静脉内自我给药。UPLC/质谱分析表明可卡因- 诱导的DA积累，而不是其他神经递质的积累，被Becn 2消耗减弱。此外，本发明还提供了一种方法， 抑制Becn 2上游的自噬激酶，包括ULK 1和Vps 34，模拟了 Becn 2缺失/突变对可卡因诱导的奖励行为和DA积累的影响，表明 在可卡因反应的调节中存在ULK 1-Vps 34-Becn 2轴。此外，生物化学 分析显示D2 R是Becn 2通过与Becn 2相关蛋白GASP 1结合而降解的靶标。 Becn 2的遗传抑制或ULK 1的药理学抑制类似地增加纹状体突触前D2 Rs。 基于这些初步数据，我们提出了研究ULK 1-Vps 34-Becn 2 自噬轴通过选择性调节D2 R自身受体内溶酶体控制可卡因滥用的脆弱性 DA神经元的运输和降解。利用遗传学、成像、生物化学、细胞学和 行为方法，我们的目标是回答以下问题：Becn 2如何在DA神经元中发挥作用， 调节可卡因摄入的获得、剂量反应、可卡因寻求的恢复和D2 R激动剂 (Aim（1）？DA神经元中是否存在ULK 1-Vps 34-Becn 2自噬通路调节这些可卡因奖赏 行为，如果是，它是如何工作的（目标2）？我们预计，有了这两个基本相关的目标， 我们的研究将建立一个以Becn 2为中心的自噬轴在D2调节中的功能和机制。 受体代谢、DA能功能和可卡因相关的奖赏行为。更好地理解本 新的分子机制可能为开发可卡因滥用的治疗方法提供新的选择， 药物滥用的类型。