Functional engraftment of stem cell-derived cortical interneurons

摘要 皮质中间神经元代表一大类抑制神经元，对控制大脑是必不可少的。 兴奋性和协调行为。抑制回路的中断与许多脑部疾病有关 精神障碍，包括癫痫、智力残疾、自闭症、精神分裂症和头部损伤。最近，进展 在小鼠和人类干细胞研究表明，多能细胞可以用来产生丰富的 体外培养的皮质神经元和中间神经元群体。然而，很少有研究系统地研究 干细胞来源的外源性抑制神经元如何作为一种细胞疗法来修改 活体内的神经回路。这个K99/R00应用程序的总体目标是确定皮质中间神经元如何 来源于小鼠和人类诱导的多能干细胞(iPS细胞)在功能上整合到 出生后的大脑。该奖项的指导阶段将在加州大学旧金山分校进行 在Scott Baraban博士的指导下，该项目将在我自己的实验室中继续进行 独立教员的职位得到了保障。在具体的目标1和2中，我将使用基于推广者的记者 IPS细胞产生的皮质间神经元前体细胞的纯化及其分化特性的构建 在体外(目标1)和移植后(目标2)使用一系列解剖、分子和 电生理学方法。在目标3(R00阶段)中，我将确定iPS细胞的连接模式- 衍生的中间神经元移植到出生后的大脑中。了解皮质中间神经元是如何从 在功能上结合到受体电路中的干细胞来源将提供关于其 功能可塑性是将这些发现转化为新的基于中间神经元的细胞的关键一步 治疗。我的长期目标是建立一个独立的致力于了解神经机制的 电路组织和开发用于脑修复和再生的新的干细胞策略，特别是 与神经元间功能障碍相关的脑部疾病。这项研究需要在STEM方面进行广泛的培训 细胞生物学，加州大学旧金山分校是完成这项申请的指导阶段的杰出机构， 主要是因为有很多杰出的神经学家和临床医生在进行神经干细胞研究 加州大学旧金山分校在干细胞领域的研究和先驱作用。除了巴拉班博士杰出的 指导，我已经组建了一个由国际公认的科学家组成的团队，他们将为我提供帮助- 在本提案的两个阶段，在技术培训、正式课程和职业指导方面。总的来说， 这些培训经历对我成功获得学术教职和 建立我的独立研究计划。

Abstract Cortical interneurons represent a broad class of inhibitory neurons that are essential for controlling brain excitability and coordinating behavior. Disruption of inhibitory circuits has been implicated in a number of brain disorders, including epilepsy, intellectual disability, autism, schizophrenia, and head injury. Recently, advances in mouse and human stem cell research suggest that pluripotent cells can be used to generate enriched populations of cortical neurons and interneurons in vitro. However, few studies have systematically examined how exogenous inhibitory neurons derived from stem cell sources might be used as a cell-therapy to modify neural circuitry in vivo. The overall goal of this K99/R00 application is to determine how cortical interneurons derived from mouse and human induced pluripotent stem cells (iPS cells) functionally incorporate into the postnatal brain. The mentored phase of the award will be conducted at University of California, San Francisco under the guidance of Dr. Scott Baraban and the project will be continuted in my own laboratory after an independent faculty position is secured. In Specific Aims 1 and 2, I will use a promoter-based reporter construct to purify coritcal interneuron precursors generated from iPS cells and characterize their differentiation in vitro (Aim 1) and after transplantation (Aim 2) using a series of anatomical, molecular, and electrophysiological approaches. In Aim 3 (R00 phase), I will determine the connectivity patterns of iPS cell- derived interneurons grafted into the postnatal brain. Understanding how cortical interneurons generated from stem cell sources functionally incorporate into the recipient circuitry will provide new information about their functional plasticity and is a critical step toward translating these findings into new interneuron-based cell therapies. My long term goal is to build an independent dedicated to understanding mechanisms of neural circuit organization and to develop novel stem cell strategies for brain repair and regeneration, particularly for brain disorders associated with interneuron dysfunction. This research will require extensive training in stem cell biology, and UCSF is an outstanding institution to complete the mentored phase of this application, primarily due to the rich community of prominent neuroscientists and clinicians performing neural stem cell research and the pioneering role of UCSF in the stem cell field. In addition to Dr. Baraban's outstanding mentorship, I have assembled a team of internationally recognized scientists who will provide me with hands- on technical training, formal coursework, and career guidance during both phases of this proposal. Overall, these training experiences will be critical for me to successfully obtain an academic faculty position and establish my independent research program.