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: 8738733
• 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 2015-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8738733
• 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

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.

描述（由申请人提供）： 皮质中间神经元代表了一大类抑制性神经元，它们对于控制大脑兴奋性和协调行为至关重要。 抑制回路的破坏与许多脑部疾病有关，包括癫痫、智力障碍、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，多能细胞可用于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如何从干细胞来源的外源性抑制神经元可能被用作细胞疗法，以修改体内的神经回路。这项K99/R 00应用的总体目标是确定来自小鼠和人类诱导多能干细胞（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弗朗西斯科博士的指导下进行，该项目将在获得独立教师职位后在我自己的实验室继续进行。在具体目标1和2中，我将使用基于启动子的报告构建体纯化从iPS细胞产生的皮质中间神经元前体，并使用一系列解剖学、分子和电生理学方法表征其在体外（目标1）和移植后（目标2）的分化。在Aim 3（R 00阶段），我将确定移植到出生后大脑中的iPS细胞衍生的中间神经元的连接模式。了解从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，并且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回路组织的机制，并开发新的干细胞策略，用于大脑修复和再生，特别是与中间神经元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的广泛培训，和UCSF是一个优秀的机构，以完成这一应用的指导阶段，主要是由于杰出的神经科学家和临床医生进行神经干细胞研究和UCSF在干细胞领域的先驱作用丰富的社区。除了巴拉班博士出色的指导外，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。总的来说，这些培训经验对我成功获得学术教师职位和建立我的独立研究计划至关重要。