A novel role for Oligodendrocyte Progenitor Cells (OPC) in opiate addiction

项目总结/摘要 海洛因成瘾是一种慢性脑部疾病，其特征是强迫性药物摄入， 存在负面后果。海洛因成瘾者终生容易复发，部分原因是 寻求毒品的动机增强。不幸的是，海洛因成瘾的药物治疗仍然有限 而且大部分无效。为了鉴定新的治疗靶点以帮助预防复发， 必须更好地理解导致复发的行为的分子机制，特别是动机。是 已知滥用药物诱导中脑边缘多巴胺系统关键区域的可塑性，包括 前额叶皮层（PFC）。具体来说，该区域内的改变与适应不良有关。 与成瘾相关的行为，包括吸毒者的动机，这可以通过使用 在人类条件和成瘾动物模型中的渐进比率测试。虽然许多细胞 海洛因诱导的适应主要集中在PFC内的神经元细胞，我们的初步研究表明， 在目的1中提供的数据已经确定了神经胶质细胞，特别是少突胶质细胞， 海洛因自我给药后，成瘾的动物模型。我已经证明了一个转录因子 在少突胶质祖细胞（OPC）分化的调节中，Sox 10在PFC中上调， 通过染色质重塑剂BRG 1自我施用海洛因后。此外，我已经确定Sox 10是一种 关键调解人在一个标志性的行为与成瘾，动机寻求药物。虽然这些 新的研究增加了我们对神经胶质转录物在成瘾状态中作用的理解， 与整个谱系的细胞相比，在祖细胞群体中特异性发生的变化仍然未知。 因此，我建议检查OPC的一种特定亚型（NG 2+细胞）的细胞变化， 与其他神经胶质细胞不同的生理特性。我在目标2中的中心假设是，海洛因诱导 与生理盐水对照相比，PFC中NG 2+细胞内的转录组学变化。我将决定 使用遗传小鼠模型组合，在该细胞亚群中基因转录的细胞类型特异性改变 使用翻译核糖体亲和性，通过病毒介导的基因转移从NG 2细胞中选择性纯化RNA 纯化（TRAP）。将使用RNA测序分析RNA的转录物变化。科学家， 该建议将确立NG 2细胞在海洛因成瘾状态中的关键作用。虽然这些研究 在进行中，我将参加一个多方面的培训计划，旨在发展必要的非板凳技能 我的最终目标是成为一名学术界的终身教职员工。此外，目标3是 旨在帮助确定一个资金充足的博士后导师谁继续帮助我发展 从博士后学者过渡到研究密集型独立研究员所需的技能 这将包括通过K99奖获得我自己的资金。

Project Summary / Abstract Heroin addiction is a chronic brain disease that is characterized by compulsive drug intake despite the presence of negative consequences. Heroin addictions have a lifelong vulnerability to relapse, in part due to heightened motivation to seek the drug. Unfortunately, pharmacotherapies for heroin addiction remain limited and largely ineffective. In order to identify novel therapeutic targets to aid in the prevention of relapse, the molecular mechanism of behaviors leading to relapse, specifically motivation, must be better understood. It is known that drugs of abuse induce plasticity in key regions of the mesolimbic dopamine system, including the prefrontal cortex (PFC). Specifically, alterations within this region have been associated with the maladaptive behaviors associated with addiction, including motivation for drug reinforcers, which can be assed using a progressive ratio test in both the human condition and animal models of addiction. While many of the cellular adaptions induced by heroin have predominately focused on the neuronal cells within the PFC, our preliminary data, presented in Aim 1, has identified a dysregulation in glial cells, and specifically oligodendrocytes, following heroin self-administration, an animal model of addiction. I have shown that a transcription factor key in the regulation of oligodendrocyte progenitor cell (OPC) differentiation, Sox10, is upregulated in the PFC following heroin self-administration via chromatin remodeler BRG1. Furthermore, I have identified Sox10 as a critical mediator in a hallmark behavior associated with addiction, motivation to seek the drug. While these novel studies have added to our understanding of the role of glial transcripts in the addicted state, the changes that are occurring specifically in progenitor population, versus cells of the whole lineage, remain unknown. Therefore, I am proposing to examine cellular changes in a specific subtype of OPCs, NG2+ cells, which have distinct physiology properties from other glial cells. My central hypothesis in Aim 2 is that heroin induces transcriptomic changes within the NG2+ cells in the PFC compared to saline controls. I will determine the cell-type specific alterations in genes transcription in this subset of cells using genetic mouse models combined with viral-mediated gene transfer to selectively purify RNA from NG2 cells using Translating Ribosomal Affinity Purification (TRAP). The RNA will be analyzed for transcript changes using RNA-sequencing. Scientifically, this proposal will establish a critical role for NG2 cells in the heroin-addicted state. While these studies are underway, I will participate in a multifaceted Training Plan designed to develop the non-bench skills necessary to reach my ultimate goals of becoming a tenured faculty in an academic setting. In addition, Aim 3 was designed to aid in the identification of a well-funded postdoctoral mentor who continue to help me develop the skills necessary to transition from a postdoctoral scholar to an independent researcher at a research intensive institution, which will include receiving my own funding through a K99 award.