Uptake2 transporters: Novel sex-dependent molecular targets to treat stimulant use disorder

Uptake2转运蛋白：治疗兴奋剂使用障碍的新型性别依赖性分子靶标

基本信息

批准号：
10595444
负责人：
LYNETTE C DAWS
金额：
$ 67.43万
依托单位：
UNIVERSITY OF TEXAS HLTH SCIENCE CENTER
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-07-15 至 2028-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10595444
关键词：
Affinity Age Amphetamine Addiction Amphetamines Astrocytes Attenuated Behavior Behavioral Brain region Chronic Cocaine Compensation Complex Corpus striatum structure Data Development Disease Dopamine Dopamine Receptor Dorsal Dose Drug Design Elements Female Gene Expression Goals Histologic In Vitro Knock-out Knockout Mice Knowledge Legal Light Literature Locomotion LoxP-flanked allele Measures Mediating Methamphetamine Modeling Molecular Target Motor Activity Mus Neuroglia Nucleus Accumbens Organic Cation Transporter Pathway interactions Persons Pharmaceutical Preparations Pharmacology Presynaptic Terminals Property Public Health Rewards Role Self Administration Sex Differences Signal Transduction Stimulant Substance Use Disorder Tamoxifen Testing Therapeutic Time Transgenic Mice Tyrosine 3-Monooxygenase Viral Wild Type Mouse addiction amphetamine use attenuation bath salts conditional knockout conditioned place preference dopamine transporter dopaminergic neuron drug of abuse effective therapy in vivo inhibitor insight knock-down male mephedrone monoamine mouse model neurotransmission nigrostriatal pathway novel presynaptic prevent protein expression psychostimulant response sex stereotypy stimulant abuse stimulant dependence stimulant use disorder synthetic drug therapeutic target treatment strategy uptake vesicular monoamine transporter

项目摘要

Medications to treat substance use disorders (SUD) exist for many drugs of abuse, but not for psychostimulants, such as amphetamine (AMPH), and methamphetamine. They are also lacking for increasingly used synthetic drugs designed to mimic the actions of known psychostimulants. Both known and new psychoactive substances continue to pose a major and increasing public health threat. To develop effective treatments, the mechanisms by which these stimulants produce their abuse-related effects need to be better understood. Many stimulants interact with the dopamine (DA) transporter (DAT), which is thought to mediate their abuse-related effects. However, strategies targeting DAT have yielded little to no benefit in the treatment of psychostimulant addiction, raising the possibility that these stimulants have significant actions elsewhere to modulate dopaminergic neurotransmission. Consistent with this, a rapidly growing literature supports a prominent role for organic cation transporter 3 (OCT3) in regulating dopaminergic neurotransmission. Our preliminary data support this idea, showing that decynium-22 (D22), an inhibitor of OCT3, inhibits DA release in vivo in male OCT3 wild-type (WT) mice, an effect lost in constitutive OCT3 knockout (KO) mice. Furthermore, in vitro studies showed that AMPH-induced substrate efflux could be inhibited by D22 in a manner independent of cocaine-sensitive transporters. These finding were paralleled by a loss of conditioned place preference (CPP) for AMPH in male mice, and an attenuation in female OCT3 KO, with D22 preventing CPP in both WT sexes. Preliminary data in male mice show that the dose-response curve for self-administration of AMPH is dramatically downward and rightward shifted in male OCT3 KO mice relative to their WT counterpart. In light of these findings, we have established and validated OCT3 floxed mice so as to examine the role of OCT3 in the absence of any developmental compensation that may occur in germline KOs. Harnessing the power of these mice, new data show that tamoxifen inducible or viral KO of OCT3 robustly attenuates AMPH- evoked DA release and CPP. Proposed studies will interrogate OCT3’s role in the ability of AMPH to impact behavior and DA efflux by testing the novel and potentially transformative hypothesis that AMPH driven DA release is not mediated solely by DAT but by reverse transport via OCT3 as well. Thus, DAT-OCT3 redundancy may explain why DAT-based therapeutics have been unsuccessful in treating AMPH use disorders. We will use a combination of pharmacology, novel and established transgenic mouse models (DA neuron-specific vs glia- specific), and brain-region specific viral knockdown, to determine the role of OCT3 in AMPH-induced behavioral effects (locomotion, rearing, stereotypy, sensitization, CPP, self-administration) (Aim 1), and AMPH-induced DA efflux and clearance in vivo (Aim 2). In Aim 3 we will determine how tamoxifen inducible KO or viral KO of OCT3 impacts key elements in DA signaling and how OCT3 adapts to chronic AMPH self-administration. These studies will fill fundamental knowledge gaps about the importance of OCT3 in abuse-related effects of AMPH, and its congeners, which will help identify novel molecular targets for medications to treat AMPH addiction.

为许多滥用药物而存在治疗药物使用障碍的药物（SUD），但没有用于精神刺激剂，例如苯丙胺（AMPH）和甲基苯丙胺。他们还缺乏越来越多的合成旨在模仿已知心理刺激剂的作用的药物。已知和新的精神活性物质继续构成重大的公共卫生威胁。为了开发有效的治疗方法，这些兴奋剂产生与滥用相关的效果需要更好地理解。许多兴奋剂相互作用多巴胺（DA）转运蛋白（DAT）被认为可以介导其与滥用相关的影响。然而，针对DAT的策略在治疗心理刺激成瘾方面几乎没有任何好处，从而提高了这些兴奋剂在其他地方采取重大行动来调节多巴胺能神经传递的可能性。与此一致，迅速增长的文献支持有机阳离子转运蛋白3（OCT3）的重要作用控制多巴胺能神经传递。我们的初步数据支持这一想法，表明Decynium-22 （D22），一种OCT3的抑制剂，抑制了雄性Oct3野生型（WT）小鼠体内DA释放，这种效应在组成型Oct3敲除（KO）小鼠。此外，体外研究表明，AMPH诱导的底物外排 D22可以以与可卡因敏感转运蛋白无关的方式抑制。这些发现是平行的雄性小鼠AMPH的条件偏好（CPP）以及女性Oct3 KO的衰减， D22可以防止CPP在两个WT性别中。雄性小鼠的初步数据表明，剂量反应曲线 AMPH的自我给药在雄性Oct3 KO小鼠相对于他们 wt对准。鉴于这些发现，我们已经建立并验证了Oct3 Floxed小鼠以检查 OCT3在没有种系KO中可能发生的任何发育补偿的情况下的作用。利用这些小鼠的力量，新的数据表明，他莫昔芬可诱导或病毒的oct3稳健降低唤起了DA释放和CPP。拟议的研究将询问Oct3在AMPH影响能力中的作用通过测试Amph驱动的DA释放的新颖而潜在的变革假设，通过测试新颖的转化假设来进行行为和DA外排不是仅由DAT介导的，而是通过OCT3进行反向传输。那是dat-oct3冗余可能解释为什么基于DAT的治疗在治疗AMPH使用障碍方面没有成功。我们将使用一个药理学，新颖和已建立的转基因小鼠模型的组合（DA神经元特异性与Glia- 具体）和大脑区域特定的病毒敲低，以确定OCT3在AMPH诱导的行为中的作用效果（运动，饲养，刻板印象，灵敏度，CPP，自我管理）（AIM 1）和AMPH诱导的DA 体内外排和清除（AIM 2）。在AIM 3中，我们将确定oct3的他莫昔芬如何诱导KO或病毒KO 影响DA信号传导中的关键要素，以及OCT3如何适应慢性Amph自我管理。这些研究将填补有关OCT3在AMPH滥用相关影响及其的重要性的基本知识差距及其同类物将有助于确定用于治疗Amph成瘾的药物的新型分子靶标。