Engineering human interneurons for neural repair

工程人类中间神经元用于神经修复

• 批准号: 10285320
• 负责人: Lyandysha Viktorovna Zholudeva
• 金额: $ 6.64万
• 依托单位: J. DAVID GLADSTONE INSTITUTES
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10285320
• 关键词: Engineering; human; interneurons; neural; repair

PROJECT SUMMARY/ABSTRACT Emerging technologies, such as cell-based repair strategies, offer new promise for some of the most devastating medical conditions that currently lack treatments. However, to harness the full therapeutic potential of stem cells, it will be necessary to understand how to direct their differentiation to appropriate cell phenotypes and ensure that their phenotype and function persist after transplantation into a pathologic environment. These gaps in knowledge are the cornerstones of my long-term training plan. My research vision is focused on creating human induced pluripotent stem cell (iPSC)-based tissues that enable the study of human central nervous circuits to accelerate the identification of therapeutic targets for neural circuit restoration in the setting of disease and injury. To do so, I will build upon my doctoral studies and publications using pre-clinical spinal cord injury as a testbed for my hypotheses regarding the therapeutic potential of transplanted human iPSC-derived neural tissue. Spinal cord injury (SCI) is a devastating condition, resulting in irreversible, life-changing disabilities. However, pre- clinical studies and clinical reports have demonstrated a remarkable innate neuroplastic potential of the injured central nervous system. Key to this neuroplasticity are spinal interneurons. Spinal interneurons are recognized as having potential for serving as synaptic relays across sites of spinal trauma and altering their activity to facilitate functional plasticity. The primary goal of this proposal is to determine if specific interneuronal sub-types generated from human stem cells can be used to restore functional connectivity in the injured nervous system. Building upon my previous work, I will use cutting edge technology to generate and phenotype human excitatory spinal interneurons to investigate formation of functional neural networks in vitro (Aim 1) and assess their phenotypic persistence and functional changes after transplantation into the intact and injured spinal cord (Aim 2). I will transplant human iPSC-derived neurons into a transgenic mouse model (excitatory and inhibitory V2a- DREADD) with SCI, which will enable functional interrogation of host-donor-host connections with the use of designer drugs. Successful completion of this project will reveal the first insight into the therapeutic potential of engineered human interneurons for neural repair and will provide the preliminary data I will use in future career development grant applications. Completing this work at Gladstone Institutes will enable me to develop an impeccable professional network to learn cellular engineering, master single cell RNA techniques for characterization, and build novel analytical workflows to visualize multi-dimensional data. More importantly, it will give me the opportunity to take advantage of resources at Gladstone and UCSF to strengthen my scientific communication skills and pursue professional development opportunities.

项目摘要/摘要 新兴技术，如基于细胞的修复策略，为一些最具破坏性的 目前缺乏治疗的医疗状况。然而，为了充分利用干细胞的治疗潜力， 有必要了解如何引导它们分化为合适的细胞表型，并确保 它们的表型和功能在移植到病理环境中后仍然存在。这些差距存在于 知识是我长期培训计划的基石。我的研究目标是创造人类 基于诱导多能干细胞(IPSC)的组织，使人类中枢神经回路的研究能够 加快确定疾病和损伤背景下神经回路修复的治疗靶点。 为了做到这一点，我将在我的博士研究和出版物的基础上，将临床前脊髓损伤作为试验床 关于移植的人IPSC来源的神经组织的治疗潜力的假设。脊椎 脊髓损伤(SCI)是一种破坏性的疾病，会导致不可逆转的、改变生活的残疾。然而，Pre- 临床研究和临床报告表明，伤者具有显著的先天神经再生潜能。 中枢神经系统。这种神经可塑性的关键是脊髓中间神经元。脊髓中间神经元是公认的 有潜力作为跨越脊柱创伤部位的突触中继器，并改变它们的活动 促进功能可塑性。这项建议的主要目标是确定特定的神经元间亚型 从人类干细胞中产生的蛋白质可以用于恢复受损神经系统的功能连接。 在我以前工作的基础上，我将使用尖端技术来产生和表现出人类的兴奋性 脊髓中间神经元在体外形成功能神经网络的研究(目标1)并评估其 移植到完整和损伤脊髓后的表型持久性和功能变化(目的 2)。我将把人类IPSC来源的神经元移植到转基因小鼠模型中(兴奋性和抑制性V2a- DREADD)和SCI，这将允许通过使用 特制毒品。这个项目的成功完成将揭示出对治疗潜力的第一次洞察 设计了用于神经修复的人类中间神经元，并将提供我在未来职业生涯中使用的初步数据 发展赠款申请。在格莱斯顿学院完成这项工作将使我能够开发一种 无懈可击的专业网络学习细胞工程，掌握单细胞RNA技术 特性，并构建新的分析工作流以可视化多维数据。更重要的是，它 将使我有机会利用格拉德斯通和加州大学旧金山分校的资源来加强我的科学研究 沟通能力强，追求职业发展机会。