Investigation of Novel Drug Targets for Stimulant Use Disorder

兴奋剂使用障碍新药靶点的研究

基本信息

批准号：
10610011
负责人：
Allison White
金额：
$ 4.87万
依托单位：
WEST VIRGINIA UNIVERSITY
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：
2023-02-01 至 2023-12-08
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10610011
关键词：
Affect Amphetamines Animals Behavior Behavioral Behavioral Paradigm Biochemical Biochemistry Brain Brain region Cardiovascular system Carrier Proteins Cells Cellular biology Cessation of life Cocaine Cocaine use disorder Cocaine withdrawal Complex Corpus striatum structure Data Disease model Dopamine Drug Targeting Educational process of instructing Evaluation Exhibits FDA approved Functional disorder Future G-Protein-Coupled Receptors Genotype Goals Grooming Incidence Individual Intervention Intravenous Investigation Knockout Mice Label Laboratories Laboratory Animals Learning Locomotion Maintenance Measures Mediating Mental disorders Mentors Methamphetamine Modeling Molecular Molecular Biology Techniques Molecular Target Mus Neurobiology Nucleus Accumbens Operative Surgical Procedures Overdose Pharmaceutical Preparations Phase Postdoctoral Fellow Procedures Process Proteins RGS Proteins Reporting Research Research Design Research Personnel Research Project Grants Resistance Role Running Self Administration Serotonin Stimulant Stress Stress and Coping Techniques Testing Tissues Tracer Training United States Wild Type Mouse Work Writing addiction anxiety-like behavior behavior test behavioral study brain tissue career cell type cocaine use design dopamine transporter druggable target experimental study graduate student improved insight monoamine mortality mouse model neuropsychiatric disorder new therapeutic target poor health outcome post-doctoral training pre-doctoral protein expression psychostimulant serotonin transporter single-cell RNA sequencing skills stimulant use stimulant use disorder tool training opportunity transcriptome treatment strategy uptake

项目摘要

PROJECT SUMMARY The incidence of stimulant use disorder (SUD) in the United States is on the rise. Notably, no FDA-approved medications exist to treat SUD. Cocaine and methamphetamine (METH) are the primary stimulants used by individuals with SUD. Because those stimulants exert their effects via monoamine transporters (MATs), interventions that modulate MAT function could be effective SUD treatments. Our group identified the brain- expressed protein RGS12 as being a modulator of monoamine transporters, and RGS proteins are thought to be druggable targets. Thus, my first aim (my dissertation research project) is to evaluate RGS12 inhibition as a potential treatment strategy for SUD using a mouse model of the disease. Our group discovered that mice lacking RGS12 function are resistant to the locomotor-activating effects of various stimulants, including 3,4- methylendedioxymethamphetamine (MDMA). My early dissertation work focused on understanding why RGS12- null mice are resistant to MDMA-induced hyperlocomotion. As MDMA’s primary target is the serotonin transporter (SERT), I hypothesized that SERT was dysregulated in RGS12-null mice. To test my hypothesis, I measured SERT expression and function using brain tissue from wild-type and RGS12-null mice. I discovered that, compared with wild-type mice, RGS12-null mice exhibited increases in SERT expression and function. Given that loss of RGS12 in mice is accompanied by increased expression and function of SERT (my finding) and the dopamine transporter (DAT; a previous graduate student’s finding), and because cocaine targets both SERT and DAT, I hypothesized that RGS12-null mice would be resistant to cocaine withdrawal-associated behaviors. To test my hypothesis, I assessed previously reported somatic signs of cocaine withdrawal in wild-type and RGS12-null mice. My results showed that while wild-type mice display increased grooming during cocaine withdrawal, RGS12-null mice do not. In the F99 training phase, I will continue my efforts to validate RGS12 as a target for SUD by measuring cocaine or METH intravenous self-administration (IVSA), the gold standard behavioral paradigm for modeling SUD in laboratory animals. I will determine if there are Rgs12 genotype- associated effects on the acquisition and maintenance of cocaine or METH self-administration, as well as on reinstatement of extinguished cocaine or METH self-administration. My second aim (my postdoctoral research direction) is to characterize the GPCR transcriptome in SUD-relevant circuitry, as GPCRs are excellent targets for modulating the activities of cells/circuits. In the K00 training phase, I will use retrograde tracers to label single cells that project to SUD-relevant brain regions. I will then identify, using single-cell RNA-seq, the GPCR transcriptome of labeled cells. Ideally, those data will reveal GPCRs that, upon activation or inactivation, will dampen dopamine release in the nucleus accumbens. The proposed research phases will provide me ample opportunities to learn new techniques. Additionally, the proposed training plan will help me improve my skills in presenting, writing, mentoring, teaching, and leading a laboratory group.

项目摘要在美国，刺激使用障碍（SUD）的事件正在上升。值得注意的是，没有FDA批准存在治疗SUD的药物。可卡因和甲基苯丙胺（甲基苯丙胺）是使用的主要兴奋剂患有SUD的人。因为这些兴奋剂通过单胺转运蛋白（MAT）执行其效果，所以调节MAT功能的干预措施可能是有效的SUD处理。我们的小组确定了大脑表示蛋白RGS12是单胺转运蛋白的调节剂，RGS蛋白被认为是成为可吸毒的目标。这是我的第一个目标（我的论文研究项目）是评估RGS12抑制作用使用该疾病的小鼠模型的SUD的潜在治疗策略。我们的小组发现小鼠缺乏 RGS12功能对各种兴奋剂的运动活化作用有抵抗力，包括3,4-- 甲基甲基甲基苯丙胺（MDMA）。我的早期论文工作的重点是理解为什么RGS12- 无效小鼠对MDMA诱导的超替代性具有抗性。因为MDMA的主要目标是5-羟色胺转运蛋白（Sert），我假设SERT在RGS12-NULL小鼠中失调。为了检验我的假设，我测量了使用野生型和RGS12-NULL小鼠的脑组织SERT表达和功能。我发现，与野生型小鼠相比，RGS12-NULL小鼠暴露于SERT表达和功能。给出小鼠中RGS12的损失是通过增加的表达和功能来实现的（我的发现）和多巴胺转运蛋白（DAT；以前的研究生发现），并且因为可卡因针对两个Sert 和DAT，我假设RGS12-NULL小鼠对可卡因戒断相关的行为有抵抗力。为了检验我的假设，我评估了先前报道的野生型可卡因戒断的躯体迹象， RGS12-NULL小鼠。我的结果表明，虽然野生型小鼠在可卡因期间显示增长增加提取，RGS12-NULL小鼠没有。在F99培训阶段，我将继续努力验证RGS12 通过测量可卡因或甲基静脉自我给药（IVSA），金标准来实现SUD的靶标用于在实验动物中建模SUD的行为范例。我将确定是否有RGS12基因型 - 对可卡因或甲基甲基甲基化的获取和维持的相关影响，以及恢复可卡因或甲基甲基化的自我管理。我的第二个目标（我的博士后研究方向）是表征与SUD相关电路中的GPCR转录组，因为GPCR是极好的目标用于调节细胞/电路的活性。在K00训练阶段，我将使用逆行示踪剂来标记单个投射到与SUD相关的大脑区域的细胞。然后，我将使用单细胞RNA-seq识别GPCR 标记细胞的转录组。理想情况下，这些数据将揭示GPCR，在激活或灭活后将在伏隔核中抑制多巴胺释放。拟议的研究阶段将为我提供足够的学习新技术的机会。此外，拟议的培训计划将帮助我提高自己的技能展示，写作，指导，教学和领导实验室小组。