Functional engraftment of stem cell-derived cortical interneurons

干细胞来源的皮质中间神经元的功能植入

• 批准号: 8618809
• 负责人: Robert F Hunt
• 金额: $ 8.8万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-20 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8618809
• 关键词: Acute; Adult; Autistic Disorder; Award; Behavior; Brain; Brain Diseases; Brain Injuries; Brain region; California; Cell Therapy; Cell Transplantation; Cell physiology; Cells; Communities; Craniocerebral Trauma; Data; Development; Electrophysiology (science); Embryo; Engraftment; Epilepsy; Faculty; Functional disorder; Future; Goals; Hippocampus (Brain); Human; Image; In Vitro; Institution; Intellectual functioning disability; Interneurons; Laboratories; Learning; Medial; Memory; Mentors; Mentorship; Methods; Molecular; Morphology; Mus; Natural regeneration; Neocortex; Neurobiology; Neurons; Newborn Infant; Optics; Parvalbumins; Pattern; Phase; Phenotype; Population; Positioning Attribute; Procedures; Property; Protocols documentation; Reporter; Research; Research Personnel; Reverse Transcriptase Polymerase Chain Reaction; Role; San Francisco; Schizophrenia; Scientist; Secure; Seizures; Slice; Source; Staging; Stem Cell Research; Stem cells; Synapses; Techniques; Telencephalon; Therapeutic; Time; Training; Translating; Transplantation; Universities; Work; base; biocytin; brain repair; career; design; experience; fetal; gamma-Aminobutyric Acid; human data; human stem cells; improved; in vivo; induced pluripotent stem cell; inhibitory neuron; migration; mind control; nerve stem cell; nervous system disorder; neural circuit; novel; patch clamp; postnatal; progenitor; programs; promoter; public health relevance; repaired; research study; skills; stem; stem cell biology; stem cell technology; way finding

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.

抽象的 皮质中间神经元代表了一类广泛的抑制性神经元，对于控制大脑至关重要 兴奋性和协调行为。抑制回路的破坏与许多大脑功能有关 疾病，包括癫痫、智力障碍、自闭症、精神分裂症和头部损伤。近期，进展 小鼠和人类干细胞研究表明，多能细胞可用于生成富集的 体外皮质神经元和中间神经元群体。然而，很少有研究系统地考察 如何使用源自干细胞来源的外源抑制性神经元作为细胞疗法来改变 体内神经回路。该 K99/R00 应用程序的总体目标是确定皮质中间神经元如何 源自小鼠和人类的诱导多能干细胞（iPS 细胞）功能性地整合到 产后大脑。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继续进行 独立教师职位得到保障。在具体目标 1 和 2 中，我将使用基于启动子的报告器 构建纯化 iPS 细胞产生的皮质中间神经元前体并表征其分化 体外（目标 1）和移植后（目标 2）使用一系列解剖学、分子学和 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产生的 干细胞来源功能性地融入受体电路将提供有关其的新信息 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 疗法。我的长期目标是建立一个独立的致力于理解神经机制的机构 电路组织并开发用于大脑修复和再生的新型干细胞策略，特别是 与中间神经元功能障碍相关的脑部疾病。这项研究需要在干细胞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该应用程序指导阶段的杰出机构， 主要归功于从事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 加州大学旧金山分校在干细胞领域的研究和先驱作用。除了巴拉班博士的杰出 在指导下，我组建了一支由国际公认的科学家组成的团队，他们将为我提供实践- 关于本提案两个阶段的技术培训、正式课程和职业指导。全面的， 这些培训经历对于我成功获得学术教职职位至关重要 建立我的独立研究计划。