Functional engraftment of stem cell-derived cortical interneurons

描述(申请人提供)：皮质中间神经元代表一大类抑制性神经元，对于控制大脑兴奋性和协调行为是必不可少的。 抑制回路的中断与许多大脑疾病有关，包括癫痫、智力残疾、自闭症、精神分裂症和头部损伤。最近，小鼠和人类干细胞研究的进展表明，多能细胞可以在体外产生丰富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来自干细胞来源的外源性抑制性神经元如何作为一种细胞疗法来改变体内的神经回路。这个K99/R00应用程序的总体目标是确定来自小鼠和人类诱导的多能干细胞(iPS细胞)的皮质中间神经元如何在功能上整合到出生后的大脑中。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继续进行，直到获得一个独立的教员职位。在特定目标1和2中，我将使用基于启动子的报告构建来纯化iPS细胞产生的皮质间神经元前体，并使用一系列解剖学、分子和电生理方法表征它们在体外(目标1)和移植后(目标2)的分化。在目标3(R00阶段)，我将确定iPS细胞来源的中间神经元移植到出生后大脑的连接模式。了解干细胞来源产生的皮质中间神经元如何在功能上整合到受体回路中，将提供有关其功能可塑性的新信息，并是将这些发现转化为新的基于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研究中心，致力于了解神经回路组织的机制，并开发新的干细胞策略来修复和再生大脑，特别是治疗与神经元间功能障碍相关的大脑疾病。这项研究将需要在干细胞生物学方面进行广泛的培训，加州大学旧金山分校是完成这一应用的指导阶段的杰出机构，这主要是由于进行神经干细胞研究的杰出神经科学家和临床医生的丰富社区，以及加州大学旧金山分校在干细胞领域的先驱作用。除了Baraban博士出色的指导外，我还组建了一个由国际公认的科学家组成的团队，他们将在本提案的两个阶段为我提供实践技术培训、正式课程和职业指导。总体而言，这些培训经验将是我成功获得学术教员职位和建立独立研究计划的关键。

DESCRIPTION (provided by applicant): 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 cortical interneuron precursors generated from iPS cells and characterize their differentiation in vitro (Aim 1) and after transplantation (Aim 2) using a serie 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 successfull obtain an academic faculty position and establish my independent research program.