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.

项目概要在美国，兴奋剂使用障碍 (SUD) 的发生率正在上升，值得注意的是，该药物尚未获得 FDA 批准。可卡因和甲基苯丙胺 (METH) 是治疗 SUD 的主要兴奋剂。因为这些兴奋剂通过单胺转运蛋白（MAT）发挥作用，调节 MAT 功能的干预措施可能是有效的 SUD 治疗方法。蛋白质将 RGS12 表达为单胺转运蛋白的调节剂，并且 RGS 蛋白质被认为因此，我的第一个目标（我的论文研究项目）是评估 RGS12 抑制作用。我们的小组使用该疾病的小鼠模型发现，小鼠缺乏 SUD 的潜在治疗策略。 RGS12 功能可抵抗各种兴奋剂的运动激活作用，包括 3,4- 我早期的论文工作重点是了解为什么 RGS12-。空白小鼠对 MDMA 诱导的过度运动有抵抗力，因为 MDMA 的主要目标是血清素转运蛋白。（SERT），我发现 RGS12 缺失小鼠中 SERT 失调。为了验证我的假设，我进行了测量。使用野生型和 RGS12 缺失小鼠的脑组织进行 SERT 表达和功能，我发现，与野生型小鼠相比，RGS12 缺失小鼠的 SERT 表达和功能有所增加。小鼠中 RGS12 的缺失伴随着 SERT 表达和功能的增加（我的发现）多巴胺转运蛋白（DAT；以前的研究生的发现），并且因为可卡因同时针对 SERT 和 DAT，我亲眼目睹 RGS12 缺失的小鼠对可卡因戒断相关行为具有抵抗力。为了验证我的假设，我评估了之前报道的野生型和可卡因戒断体征。我的研究结果表明，野生型小鼠在吸食可卡因后表现出更多的梳理行为。在 F99 训练阶段，我将继续努力验证 RGS12 作为一种药物。通过测量可卡因或冰毒静脉自我给药 (IVSA)（黄金标准）来确定 SUD 的目标在实验动物中建模 SUD 的行为范式我将确定是否存在 Rgs12 基因型。对可卡因或冰毒自我给药的获取和维持的相关影响，以及恢复已熄灭的可卡因或冰毒自我管理我的第二个目标（我的博士后研究。方向）是为了表征 SUD 相关电路中的 GPCR 转录组，因为 GPCR 是极好的目标在K00训练阶段，我将使用逆行示踪剂来标记单个细胞/电路的活动。然后，我将使用单细胞 RNA-seq 来识别投射到 SUD 相关大脑区域的细胞。理想情况下，这些数据将揭示 GPCR，这些 GPCR 在激活或失活后会发生变化。抑制伏隔核中的多巴胺释放。拟议的研究阶段将为我提供充足的信息。此外，拟议的培训计划将帮助我提高我的技能。展示、写作、指导、教学和领导实验室小组。