Developing an Ex Vivo Model of the Mesolimbic Pathway for Studying Addiction Phenotypes

项目摘要 吸毒和吸毒成瘾仍然是一个日益严重的重大社会问题。它是在反复使用药物的过程中发展起来的 这会导致一系列分子、转录和表观遗传修饰，重塑大脑中的神经元 中脑边缘通路引导成瘾行为。尽管进行了广泛的研究，但治疗选择在一定程度上仍然有限 因为导致上瘾行为的潜在机制仍然不完全清楚。最新版本 研究潜在分子机制的研究已经在动物模型中进行，特别是在小鼠和大鼠身上， 这使得科学家能够操纵特定的变量，并评估药物寻找反应的变化。尽管 尽管这些模型的威力很大，但小鼠的组织生理分子与人类的相关性有多大程度上的局限性。 例如，它们各自在数量、细胞类型组成和功能方面存在显著差异 中脑边缘通路神经元。因此，需要体外模型系统来捕捉人类的基因， 中边缘细胞类型和连接的表观遗传学、转录和多细胞特性。我们将为 这一目标是通过将人类干细胞分化为含有多巴胺能神经元和中等刺神经元的有机体来实现的 中脑边缘通路，并在两者之间产生功能性突触。模特将通过揭露 培养神经元对可卡因的反应，研究它们的分子、转录和表观遗传学反应，并进行比较 对可用的老鼠和人类数据的反应。有了这个模型，我们可以进一步了解关键的潜在机制 这有助于形成成瘾，可能导致药物开发的特定目标。

Project Summary Drug abuse and addiction continues to be a major and growing societal problem. It develops during repeated drug use which causes a series of molecular, transcriptomic, and epigenetic modifications which remodel the neurons in the mesolimbic pathway leading addictive behavior. Despite extensive studies, treatment options remain limited in part because the underlying mechanisms contributing to addictive behavior are still not fully understood. Most previous research to study underlying molecular mechanisms have been performed in animal models specifically mice and rats, which allow scientists to manipulate a specific variable and assess alterations in drug seeking responses. Despite the power of these models, there are limitations to how well mice molecular through tissue physiologies correlate to humans. For example, there are significant differences in the number, cell type compositions, and functions of their respective mesolimbic pathway neurons. Therefore, there is a need for ex vivo model systems which capture the human genetic, epigenetic, transcriptional, and multicellular properties of mesolimbic cell types and connections. We will contribute to this goal by differentiating human stem cells into organoids containing dopaminergic and medium spiny neurons of the mesolimbic pathway and generate functional synapses between the two. The model will be interrogated by exposing the cultured neurons to cocaine and studying their molecular, transcriptional, and epigenetic responses and comparing these responses to available mice and human data. With this model, we can further understand the key underlying mechanisms that contribute to addiction formation potentially leading to specific targets for drug development.