Molecular Neurobiology of Human Opioid Use Disorder

项目摘要 阿片类药物成瘾是一种全国性的流行病，导致了美国历史上最致命的药物过量危机， 伴随着由于滥用海洛因和阿片类处方药而造成的过度医疗负担。 仍然缺乏关于阿片类药物使用障碍的神经生物学知识来推动新的治疗方法 迫切需要为目前主要基于阿片类药物的药物提供选择， 自己承担责任。我们的反向翻译研究工作的一个基本核心是填补关键的 通过直接调查人类海洛因滥用者的大脑来填补知识空白。通过这样的策略， 最近发现了一种以前未被认识到的阿片类药物的神经生物学影响，表明阿片类药物诱导的 与FYN相关的表观遗传改变，其介导谷氨酸能信号传导并磷酸化tau（pTau）; 我们还在海洛因滥用者中检测到显著的谷氨酸能改变和高pTau病理学。FYN是一个Src 突触后致密物内的一种酪氨酸激酶家族，使tau蛋白磷酸化，参与微管稳定性 和动力学，因此是细胞骨架重塑的调节剂，这是成瘾的关键特征。我们验证 在自我施用海洛因的大鼠和我们的慢性阿片类药物中，FYN靶向Tau磷酸化水平升高。 体外细胞培养模型。此外，我们能够抑制海洛因自我管理（SA）和寻求行为 在用Fyn抑制剂和Fyn敲低治疗的动物中。这些多学科和综合数据提供了 一个强大的基础上，询问FYN在阿片类药物滥用与治疗发展的目标。我们 假设Fyn上调及其导致的中皮质边缘脑中的下游靶向损伤 这些区域有助于海洛因成瘾行为，可以作为治疗干预的目标。我们建议（1） 确定与海洛因相关的中皮质边缘区FYN相关网络的分子特征 在人类和大鼠海洛因SA模型中以细胞特异性方式通过RNA测序确定滥用，以及（2）表征 Fyn介导的细胞特异性下游改变是海洛因SA和寻求行为的基础。的 下游机制利用病毒介导的Fyn对细胞内信号级联的细胞特异性作用 和pTau以及细胞骨架组织。从我们的综合多学科研究中获得的结果将 推进阿片类药物滥用相关的Fyn相关异常的知识，并提供基于科学的 药理学的目标是扩大阿片类药物成瘾的治疗选择。

Project Summary Opioid addiction is a national epidemic contributing to the deadliest drug overdose crisis in US history and accompanied by excessive healthcare burdens due to the misuse of heroin and opioid prescription medications. There continues to be a lack of neurobiological knowledge about opioid use disorder to drive novel therapies critically needed to provide options to the current medications that are predominantly opioid-based and thus of abuse liability themselves. A fundamental core of our reverse translational research efforts has been to fill critical gaps of knowledge by direct investigation of the brains of human heroin abusers. Through such strategies we recently discovered a previously unrecognized neurobiological impact of opioids suggesting opioid-induced epigenetic alteration linked to the FYN which mediates glutamatergic signaling and phosphorylates tau (pTau); we also detected significant glutamatergic alterations and hyper-pTau pathology in heroin abusers. FYN is a Src family tyrosine kinase within the postsynaptic density that phosphorylates tau, involved in microtubule stability and dynamics, and thus a regulator of cytoskeletal remodeling which is a key feature of addiction. We verified elevation of FYN-targeted Tau phosphorylation in rats that self-administered heroin and in our chronic opioid in vitro cell culture model. Moreover, we were able to inhibit heroin self-administration (SA) and seeking behaviors in animals treated with a Fyn inhibitor and Fyn knockdown. These multidisciplinary and integrative data provides a strong foundation on which to interrogate FYN in opioid abuse with the goal of therapeutic development. We hypothesize that upregulation of Fyn and its resulting downstream target impairments in mesocorticolimbic brain areas contribute to heroin addiction behavior and can be targeted for treatment interventions. We propose to (1) determine the molecular signature of FYN-related networks in mesocorticolimbic regions associated with heroin abuse by RNA-sequencing in a cell-specific manner in human and the rat heroin SA model and to (2) characterize cell-specific downstream alterations mediated by Fyn underlying heroin SA and seeking behaviors. The downstream mechanisms leverage viral mediated cell-specific effects of Fyn on intracellular signaling cascades and pTau as well as cytoskeletal organization. Results gained from our integrative multidisciplinary study will advance knowledge of Fyn-related abnormalities underlying opioid abuse and provide science-based pharmacotherapeutic targets to expand treatment options for opioid addiction.