Pathway-specific Intervention in Prelimbic Cortical Circuitry Decreases Cocaine-seeking

对前额皮质回路进行特定通路干预可减少可卡因寻求

• 批准号: 10268963
• 负责人: Jacqueline F. McGinty
• 金额: $ 33.87万
• 依托单位: MEDICAL UNIVERSITY OF SOUTH CAROLINA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-30 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10268963
• 关键词: Abstinence; Anxiety; Brain-Derived Neurotrophic Factor; CREB1 gene; Caliber; Cocaine; Cocaine Dependence; Coupled; Cues; Cyclic AMP; Cyclic AMP-Dependent Protein Kinases; Data; Dendritic Spines; Drug abuse; Early Intervention; Electrophysiology (science); Environment; Extracellular Signal Regulated Kinases; Female; Functional disorder; Future; Glutamates; Head; Image; Individual; Infusion procedures; Intervention; Mediating; Nature; Neurobiology; Neurons; Pathway interactions; Performance; Pharmaceutical Preparations; Phase; Phosphorylation; Pre-Clinical Model; Prefrontal Cortex; Protein Dephosphorylation; Protein Kinase A Inhibitor; Proteins; Rattus; Regulation; Relapse; Research; Rewards; Role; Signal Transduction; Slice; Structure; Structure of paraventricular nucleus of thalamus; Substance Use Disorder; Synaptic plasticity; Testing; Time; Viral Vector; Withdrawal; Woman; addiction; anxiety-related behavior; cocaine self-administration; combinatorial; designer receptors exclusively activated by designer drugs; effective therapy; insight; male; men; neuroadaptation; new therapeutic target; novel; pre-clinical; prevent; relating to nervous system; retrograde transport; transmission process; withdrawal-induced anxiety

Specific Aims A major challenge for individuals suffering from substance use disorders (SUDs) is the lack of effective treatments that reduce relapse vulnerability. Drug predictive cues in the environment are powerful triggers for relapse and understanding how these persistent associations are formed and maintained is a critical focus of preclinical SUD research. In this proposal, we have built on our discovery that immediately after the end of cocaine self administration (SA), an infusion of brain-derived neurotrophic factor (BDNF) into the prelimbic (PL) prefrontal cortex prevents the cocaine SA-induced dephosphorylation of key glutamatergic-related plasticity- related proteins (PRPs), including GluN2A, GluN2B, ERK MAP kinase, and CREB32,96. This early intervention with BDNF also prevents prolonged cocaine-induced deficits in PL-NA core glutamatergic transmission that promote subsequent cocaine seeking9. In contrast, by the end of the first week of abstinence, protein kinase A (PKA)-dependent augmentation of GluA1 and CREB phosphorylation emerges. At that time, intra-PL BDNF has no effect on relapse8 but intra-PL infusion of a PKA inhibitor, Rp-cAMPs, reverses the hyperphosphorylation and decreases relapse70,92. More recently, we have shown that the biphasic changes in GluA1 and pCREB within the first week of abstinence are associated with similar biphasic changes in the head diameters (dH) of dendritic spines of PL–NA core neurons89. These data have spurred the overall hypothesis that interventions to decrease drug seeking must be tailored to the dynamic changes in neuroadaptations that emerge during different phases of the addiction cycle. In chemogenetic studies to investigate the contribution of specific pathways originating in PL cortex to drug seeking, we discovered that PL-NA core and PL-posterior paraventricular thalamic nucleus (pPVT) pathways oppose each others’ effects during early withdrawal. Selective cre-dependent DREADD inhibition of PL-NA core neurons infected with a retrogradely transported cre-AAV immediately after cocaine SA has no effect by itself, but reverses the suppressive effect of intra-PL BDNF on subsequent drug seeking31. In contrast, selective inhibition of the PL-pPVT pathway immediately after cocaine SA decreases subsequent cocaine- seeking, an effect that is prevented by intra-PL BDNF31. Interestingly, we also found that selective inhibition of the PL-pPVT pathway reduces anxiety-related behavior in rats withdrawing from cocaine and inactivation of pPVT decreases conditioned aversion to cocaine, suggesting that cocaine’s engagement of anxiety- and aversion-related circuitry that contributes to drug seeking includes pPVT. Taken together, our new findings suggest the novel hypothesis that cocaine SA produces differential regulation of PL-NA core and PL-pPVT pathways and that these two distinct circuits conspire to support future drug seeking. In this proposal, we will use pathway-specific, combinatorial chemogenetic approaches, slice electrophysiology, and state-of-the-art dendritic spine and PRP imaging in PL-NA core and PL-pPVT circuitry to explore the time-dependent plasticity occurring in this network that influences cocaine seeking and aversion. We will use these approaches in the following aims that will be performed in male and female rats. Aim 1. BDNF regulation of PRPs and structural and synaptic plasticity in PL-NA core vs. PL-pPVT neurons during early withdrawal. Using viral vector approaches, we will investigate the effects of intra-PL BDNF infusion on (A) dendritic spine plasticity and PRP changes and (B) synaptic plasticity in PL-NA core vs. PL-PVT neurons during early withdrawal. Aim 2. PKA regulation of PRPs and structural and synaptic plasticity in PL-NA core vs. PL-pPVT neurons after 7 and 30 days of abstinence. (A) We will investigate the effects of inhibiting cAMP/PKA signaling on (A) dendritic spine plasticity and PRP changes and (B) synaptic plasticity in PL-NA core vs. PL- PVT neurons after 7 or 30 days of abstinence with or without relapse testing. Aim 3. The role of PL-NA core and PL-pPVT neurons in cocaine-induced anxiety and aversion in relationship to cocaine seeking after abstinence. We will use a retro-Gi DREADD approach to test the effects of inhibiting PL-NA core or PL-pPVT neurons (A) on withdrawal-induced anxiety immediately after the end of cocaine SA and (B) on conditioned avoidance of cocaine in an operant runway task before or after cocaine SA followed by cue-induced relapse testing. Scientific Impact: These studies will expand our understanding of how to reverse critical neuroadaptations underlying prefrontal dysfunctions during different phases of the addiction cycle that trigger relapse to drug- seeking. By understanding temporally dynamic, differential neuroadaptations in the PL networks regulating reward and aversion, we may be able to discover novel therapeutic targets to decrease drug-seeking.

具体目标 对于患有物质使用障碍 (SUD) 的个体来说，一个主要挑战是缺乏有效的治疗方法 减少复发脆弱性的治疗。环境中的药物预测线索是强有力的触发因素 复发和理解这些持久的关联是如何形成和维持的，是研究的一个关键焦点 临床前SUD研究。在这项提案中，我们的发现是在 可卡因自我给药 (SA)，将脑源性神经营养因子 (BDNF) 输注到边缘前 (PL) 前额皮质可防止可卡因 SA 诱导的关键谷氨酸能相关可塑性的去磷酸化 相关蛋白 (PRP)，包括 GluN2A、GluN2B、ERK MAP 激酶和 CREB32,96。这种早期干预 BDNF 还可以防止可卡因引起的 PL-NA 核心谷氨酸传输的长期缺陷 促进随后的可卡因寻找9。相比之下，在禁欲第一周结束时，蛋白激酶 A (PKA) 依赖性 GluA1 和 CREB ​​磷酸化增强出现。那时，PL内BDNF 对复发没有影响8，但 PKA 抑制剂 Rp-cAMP 的 PL 内输注可逆转 过度磷酸化并减少复发70,92。最近，我们已经表明，双相变化 禁欲第一周内的 GluA1 和 pCREB ​​与头部的类似双相变化相关 PL-NA 核心神经元树突棘的直径（dH）89。这些数据激发了总体假设 减少药物寻求的干预措施必须适应神经适应的动态变化 出现在成瘾周期的不同阶段。 在化学遗传学研究中，研究源自 PL 皮层的特定途径对药物的贡献 通过寻找，我们发现 PL-NA 核心和 PL-室旁丘脑核 (pPVT) 通路 早期戒断期间相互对抗对方的影响。 PL-NA 的选择性 cre 依赖性 DREADD 抑制 可卡因 SA 没有作用后，立即用逆行转运的 cre-AAV 感染核心神经元 本身，但逆转了 PL 内 BDNF 对后续药物寻求的抑制作用31。相比之下， 可卡因 SA 后立即选择性抑制 PL-pPVT 通路可减少随后的可卡因- 寻找，这种效应被 PL 内 BDNF31 所阻止。有趣的是，我们还发现选择性抑制 PL-pPVT 通路可减少可卡因戒断大鼠的焦虑相关行为和失活 pPVT 降低了对可卡因的条件性厌恶，表明可卡因参与了焦虑和 有助于寻求药物的厌恶相关回路包括 pPVT。 总而言之，我们的新发现提出了新的假设，即可卡因 SA 会产生差异 PL-NA 核心和 PL-pPVT 通路的调节，这两个不同的电路共同支持未来 寻求药物。在本提案中，我们将使用特定途径的组合化学遗传学方法、切片 PL-NA 核心和 PL-pPVT 电路中的电生理学、最先进的树突棘和 PRP 成像 探索该网络中发生的影响可卡因寻求和厌恶的时间依赖性可塑性。 我们将在雄性和雌性大鼠中使用这些方法来实现以下目标。 目标 1. PL-NA 核心与 PL-pPVT 中 BDNF 对 PRP 以及结构和突触可塑性的调节 早期戒断期间的神经元。使用病毒载体方法，我们将研究 PL 内的影响 BDNF 输注对 (A) 树突棘可塑性和 PRP 变化以及 (B) PL-NA 核心与非核心突触可塑性的影响 早期戒断期间的 PL-PVT 神经元。 目标 2. PL-NA 核心与 PL-pPVT 中 PRP 以及结构和突触可塑性的 PKA 调节 禁欲 7 天和 30 天后的神经元。 (A) 我们将研究抑制 cAMP/PKA 的效果 PL-NA 核心与 PL- 中 (A) 树突棘可塑性和 PRP 变化以及 (B) 突触可塑性的信号传导 禁欲 7 或 30 天后进行或不进行复发测试的 PVT 神经元。 目标 3. PL-NA 核心和 PL-pPVT 神经元在可卡因诱导的焦虑和厌恶中的作用 与寻求戒断的可卡因的关系。我们将使用 Retro-Gi DREADD 方法来测试 抑制 PL-NA 核心或 PL-pPVT 神经元 (A) 对戒断后立即引起的焦虑的影响 结束可卡因 SA 和 (B) 在操作性跑道任务之前或之后条件性避免使用可卡因 可卡因 SA 随后进行提示诱发的复发测试。 科学影响：这些研究将扩大我们对如何逆转关键神经适应的理解 在成瘾周期的不同阶段，潜在的前额叶功能障碍会引发药物复发 寻求。通过了解 PL 网络中的时间动态、差异神经适应调节 奖励和厌恶，我们也许能够发现新的治疗靶点来减少药物寻求。