Rapid and sensitive in vivo detection of opioid peptides

项目摘要 这项研究的目的有两个：更好地了解强啡肽在芬太尼戒断和 开发创新、新颖的方法，以便在体内检测内源性释放的阿片类药物。一种能力 为了更好地了解强啡肽是如何驱动与药物戒断相关的负面影响的，在 减少复发和服药过量的发生率，并使我们能够考虑长期更有效的替代方案 治疗上瘾的方法。美国卫生部表示，42249人死于过量服用阿片类药物 2016年，210万人患有阿片类药物使用障碍，这种流行病造成的经济损失为5040亿美元， 但随着受灾人数的增加，这一数字仍在上升。原因是多方面的，但包括一个 对阿片类药物如何改变大脑回路以导致止痛--这是好的，但也是什么--缺乏深入的了解 导致上瘾的大脑回路发生了变化，这是不好的。我们也知道痛苦和上瘾是并存的。 其他精神健康疾病，如焦虑、抑郁和压力，这使得研究具有凝聚力 很有挑战性。这项提议的第一个目的是直接测量体内核内强啡肽水平的变化。 芬太尼停药期间的伏安奈德。芬太尼在体内的效力是吗啡的80倍；它的快速 开始行动不仅会增加成瘾的风险，而且可能会增加戒断症状的严重程度。 我们将使用液相色谱/质谱仪，这将允许在亚fmol范围内进行检测。第二 目的关注微免疫电极的发展，以实现更时空可控的检测 阿片肽在体内的释放。这项技术将使我们能够在一秒到一分钟内检测到多肽的变化 时间刻度。这些技术的结合将第一次使我们能够检测到 体内的阿片肽。重要的是，这将有助于我们更好地治疗成瘾的知识和能力，但 也非常适用于一般的多种疾病状态和神经肽生物学。

Project Summary The goals of this research are two fold: to better understand the role of dynorphin during fentanyl withdrawal and to develop innovative, novel approaches to allow in vivo detection of endogenously release opioids. The ability to better understand how dynorphin drives negative affect associated with drug withdrawal will be critical in reducing the incidence of relapse and overdose and allow us to consider alternative more affective long-term treatments for addiction. The US Department of Health states that 42,249 people died from overdosing on opioids in 2016, 2.1 million people had an opioid use disorder and the economic cost of this epidemic was $504 billion, but is still rising along with the number of people afflicted. The reasons for this are multifaceted, but include a deep lack of understanding of how opioids alter brain circuitry to cause analgesia- which is good, but also what happens to brain circuitry to drive addiction- which is bad. We also know that pain and addiction are co-morbid with other mental health diseases such as anxiety, depression and stress, which makes cohesive research challenging. The first aim of this proposal will directly measure in vivo changes in dynorphin levels in the nucleus accumbens during withdrawal from Fentanyl. Fentanyl is 80 times more potent than morphine in vivo; its rapid onset of action not only increases the risk of addiction but likely increases the severity of withdrawal symptoms. We will use liquid chromatography/mass spectrometry which will allow detection in sub-fmol range. The second aim focuses on the development of micro-immunoelectrodes to allow more spatiotemporally controlled detection of opioid peptide release in vivo. This technology will allow us to detect peptides changes on a second to minute timescale. The combination of these techniques will, for the first time, allow us to detect dynamic changes in opioid peptides in vivo. Importantly, this will facilitate our knowledge and ability to better treat addiction, but is also very much applicable to multiple disease states and neuropeptides biology in general.