Psychostimulant use disorder is a major public health issue, and despite the scope of the problem there are currently no Food and Drug Administration (FDA)-approved treatments. There would be tremendous utility in development of a treatment that could help patients both achieve and maintain abstinence. Previous work from our group has identified granulocyte-colony stimulating factor (G-CSF) as a neuroactive cytokine that alters behavioral response to cocaine, increases synaptic dopamine release, and enhances cognitive flexibility. Here, we investigate the role of G-CSF in affecting extinction and reinstatement of cocaine-seeking and perform detailed characterization of its proteomic effects in multiple limbic substructures. Male Sprague Dawley rats were injected with PBS or G-CSF during (1) extinction or (2) abstinence from cocaine self-administration, and drug seeking behavior was measured. Quantitative assessment of changes in the proteomic landscape in the nucleus accumbens (NAc) and medial prefrontal cortex (mPFC) were performed via data-independent acquisition (DIA) mass spectrometry analysis. Administration of G-CSF during extinction accelerated the rate of extinction, and administration during abstinence attenuated cue-induced cocaine-seeking. Analysis of global protein expression demonstrated that G-CSF regulated proteins primarily in mPFC that are critical to glutamate signaling and synapse maintenance. Taken together, these findings support G-CSF as a viable translational research target with the potential to reduce drug craving or seeking behaviors. Importantly, recombinant G-CSF exists as an FDA-approved medication which may facilitate rapid clinical translation. Additionally, using cutting-edge multiregion discovery proteomics analyses, these studies identify a novel mechanism underlying G-CSF effects on behavioral plasticity.

SIGNIFICANCE STATEMENT Pharmacological treatments for psychostimulant use disorder are desperately needed, especially given the disease's chronic, relapsing nature. However, there are currently no Food and Drug Administration (FDA)-approved pharmacotherapies. Emerging evidence suggests that targeting the immune system may be a viable translational research strategy; preclinical studies have found that the neuroactive cytokine granulocyte-colony stimulating factor (G-CSF) alters cocaine reward and reinforcement and can enhance cognitive flexibility. Given this basis of evidence we studied the effects of G-CSF treatment on extinction and reinstatement of cocaine seeking. We find that administration of G-CSF accelerates extinction and reduces cue-induced drug seeking after cocaine self-administration. In addition, G-CSF leads to downregulation of synaptic glutamatergic proteins in medial prefrontal cortex (mPFC), suggesting that G-CSF influences drug seeking via glutamatergic mechanisms.

精神兴奋剂使用障碍是一个重大的公共卫生问题，尽管该问题的范围很广，但目前还没有食品和药物管理局 (FDA) 批准的治疗方法。开发一种可以帮助患者实现和保持戒断的治疗方法将具有巨大的实用性。我们小组之前的工作已将粒细胞集落刺激因子 (G-CSF) 确定为一种神经活性细胞因子，可改变对可卡因的行为反应、增加突触多巴胺释放并增强认知灵活性。在这里，我们研究了 G-CSF 在影响可卡因寻求的消退和恢复中的作用，并对其在多个边缘亚结构中的蛋白质组学效应进行了详细表征。雄性 Sprague Dawley 大鼠在 (1) 消退或 (2) 可卡因自我给药戒断期间注射 PBS 或 G-CSF，并测量药物寻求行为。通过数据独立采集 (DIA) 质谱分析对伏隔核 (NAc) 和内侧前额叶皮层 (mPFC) 中蛋白质组学景观的变化进行定量评估。G-CSF 在灭绝期间的管理加速了灭绝的速度，并且在禁欲期间管理减弱了线索诱导的可卡因寻求。对整体蛋白质表达的分析表明，G-CSF 主要调节 mPFC 中的蛋白质，这些蛋白质对谷氨酸信号传导和突触维持至关重要。总之，这些发现支持 G-CSF 作为一个可行的转化研究目标，有可能减少对药物的渴望或寻求行为。重要的是，重组 G-CSF 作为 FDA 批准的药物存在，可以促进快速临床转化。此外，使用尖端的多区域发现蛋白质组学分析，这些研究确定了 G-CSF 对行为可塑性影响的新机制。

重要性声明迫切需要对精神兴奋剂使用障碍进行药物治疗，尤其是考虑到该疾病的慢性、复发性。然而，目前还没有食品和药物管理局 (FDA) 批准的药物疗法。新出现的证据表明，以免疫系统为目标可能是一种可行的转化研究策略；临床前研究发现，神经活性细胞因子粒细胞集落刺激因子 (G-CSF) 可改变可卡因奖励和强化，并可增强认知灵活性。鉴于这一证据基础，我们研究了 G-CSF 治疗对可卡因寻求的消退和恢复的影响。我们发现 G-CSF 的给药加速了消退并减少了可卡因自我给药后线索诱导的药物寻找。此外，