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

项目摘要 药物滥用和成瘾仍然是一个日益严重的重大社会问题。它是在反复吸毒时形成的 引起一系列的分子、转录组和表观遗传修饰， 导致成瘾行为的中脑边缘通路。尽管进行了广泛的研究，但治疗选择仍然有限， 因为导致成瘾行为的潜在机制仍然没有完全被理解。大多数先前 已经在动物模型特别是小鼠和大鼠中进行了研究潜在分子机制的研究， 它允许科学家操纵一个特定的变量，并评估药物寻求反应的变化。尽管 尽管这些模型的能力有限，但小鼠的分子通过组织生理学与人类的相关性仍存在局限性。 例如，它们各自的细胞在数量、细胞类型组成和功能上存在显著差异。 中脑边缘通路神经元因此，需要一种捕获人类遗传， 中边缘细胞类型和连接的表观遗传、转录和多细胞特性。我们将有助于 这一目标是通过将人类干细胞分化成含有多巴胺能和中等多刺神经元的类器官来实现的。 mesolimbic途径，并在两者之间产生功能性突触。将通过暴露 培养的神经元可卡因和研究他们的分子，转录和表观遗传反应，并比较这些 对现有小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， 这有助于成瘾的形成，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.