Application of optogenetics in iPS cell transplantation therapy for ischemic stro

光遗传学在iPS细胞移植治疗缺血性卒中中的应用

基本信息

批准号：
9067533
负责人：
ROBERT E GROSS
金额：
$ 34.13万
依托单位：
EMORY UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-09-01 至 2019-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9067533
关键词：
Address Adult Animal Model Animals Attention Autologous Transplantation Basic Science Biological Neural Networks Biophysics Brain Cations Cell Differentiation process Cell Survival Cell Transplantation Cell Transplants Cells Cessation of life Clinical Clinical Research Combined Modality Therapy Coupled Cultured Cells Data Development Dose Electrophysiology (science)Embryo Environment Ethical Issues Goals Growth Health Human Hypoxia Imaging Techniques In Vitro Investigation Ischemia Ischemic Stroke Laboratory Research Lasers Life Light Luciferases Maps Measurement Measures Membrane Methods Modeling Molecular Molecular and Cellular Biology Mus Natural regeneration Nature Neurodegenerative Disorders Neuronal Differentiation Neuronal Injury Neurons Neurosciences Pathway interactions Patients Peripheral Pluripotent Stem Cells Property Proteins Recovery of Function Replacement Therapy Research Rodent Slice Somatic Cell Specificity Stem cell transplant Stem cells Stimulus Stroke Structure Survival Rate Techniques Technology Testing Thalamic structure Transplantation Vibrissae angiogenesis barrel cortex base brain cell brain tissue cell type chemical genetics clinical application clinically relevant coelenterazine disability effective therapy embryonic stem cell functional restoration genetic approach implantation improved in vivo induced pluripotent stem cell injured innovation membrane activity mouse model nerve stem cell neural circuit neural precursor cell neurogenesis neurovascular novel novel strategies optical imaging optogenetics post stroke pre-clinical preconditioning protective effect reconstruction regenerative regenerative therapy rehabilitation strategy repaired response restoration somatosensory stem stem cell therapy stroke therapy stroke treatment tissue repair tool

项目摘要

DESCRIPTION (provided by applicant): Stroke remains a leading cause of human death and disability while very few effective treatments are available for stroke patients. Stem cell transplantation therapy provides the possibility to regenerate and repair damaged brain tissues after ischemic stroke. The investigation takes a comprehensive and unprecedented approach to promote both trophic supports as well as cell replacement potential of pluripotent stem cells to develop a highly effective stem cell therapy for ischemic stroke. We propose that enhancing the survival and regenerative properties of transplanted cells as well as an improved host environment are critical for a successful stem cell stroke therapy. To reach this goal, our previous and preliminary studies have demonstrated a marked protective effect and increased functional benefits of combining hypoxia preconditioning (HP) and other regenerative strategies including optogenetic techniques and up regulated multiple trophic factors in the ischemic brain promoted by peripheral stimulation. Our central hypothesis is that a combination strategy of HP-primed NPCs subjected to optogenetic manipulations and improved host environment will allow better survival of transplanted as well as endogenous cells, enhance neurogenesis/angiogenesis via both exogenous and endogenous mechanisms, and results in optimal tissue repair and functional recovery after stroke. In neural progenitor cells (NPCs) derived from mouse induced pluripotent stem (iPS) cells, we will express the blue light- sensitive channelrhodopsin (ChR) channels and test the possibility that activation of ChRs by blue light stimuli or by the luciferase/ChR proten (luminopsis) substrate coelenterazine (CTZ) is a feasible and effective method to improve and evaluate neuronal differentiation, integration into host neural networks and neuronal connections after transplantation into the ischemic brain. We will examine the strategies to promote tissue repair and provide evidence for the morphological and functional restoration of ischemic brain structures in the unique barrel cortex ischemic stroke model of mice. We will demonstrate the feasibility and benefits of expression/activation of ChR channels in iPS-NPCs in vitro (Specific Aim 1) and after implantation into the post-ischemic barrel cortex (Aim 2). Based on the well-defined whisker- thalamus-barrel cortex pathway, structural and functional restoration of disrupted whisker-barrel activities will be evaluated usin a combination of cell specific and neuronal pathway specific measurements, including optogenetic, electrophysiological and optical imaging recordings (Aim 3). The proposal is from three research laboratories with complementary expertise in biophysics, electrophysiology, cellular/molecular biology and clinical neurosciences. The demonstration of cellular and tissue repairing benefits of iPS-NPCs in a particular brain structure is critical for the development of mechanism based cell transplantation therapy and the strategies will have great impacts on pre-clinical and clinical studies.

描述（由申请人提供）：中风仍然是人类死亡和残疾的主要原因，而针对中风患者的有效治疗方法却很少。干细胞移植疗法提供了再生和修复缺血性中风后受损脑组织的可能性。该研究采用全面且前所未有的方法来促进多能干细胞的营养支持和细胞替代潜力，以开发一种针对缺血性中风的高效干细胞疗法。我们认为，增强移植细胞的存活和再生特性以及改善宿主环境对于干细胞中风治疗的成功至关重要。为了实现这一目标，我们之前和初步的研究已经证明，将缺氧预处理（HP）和其他再生策略（包括光遗传学技术和外周刺激促进的缺血性大脑中的多种营养因子上调）相结合具有显着的保护作用和增加的功能益处。我们的中心假设是，HP引发的NPC经过光遗传学操作和改善宿主环境的组合策略将允许移植细胞和内源细胞更好地存活，通过外源和内源机制增强神经发生/血管生成，并导致最佳的组织修复以及中风后的功能恢复。在源自小鼠诱导多能干 (iPS) 细胞的神经祖细胞 (NPC) 中，我们将表达蓝光敏感通道视紫红质 (ChR) 通道，并测试蓝光刺激或荧光素酶/ChR 蛋白激活 ChR 的可能性。（luminopsis）基质腔肠素（CTZ）是一种可行且有效的方法，用于改善和评估移植到宿主神经网络后的神经元分化、整合到宿主神经网络和神经元连接。大脑缺血。我们将研究促进组织修复的策略，并为独特的小鼠桶状皮层缺血性中风模型中缺血性脑结构的形态和功能恢复提供证据。我们将展示 iPS-NPC 体外（具体目标 1）和植入缺血后桶状皮层（目标 2）后表达/激活 ChR 通道的可行性和益处。基于明确的胡须-丘脑-桶皮层通路，将结合细胞特异性和神经元通路特异性测量，包括光遗传学、电生理学和光学成像记录，评估受损胡须桶活动的结构和功能恢复（目标 3））。该提案来自三个研究实验室，在生物物理学、电生理学、细胞/分子生物学和临床神经科学方面拥有互补的专业知识。证明 iPS-NPC 在特定大脑结构中的细胞和组织修复功效对于开发基于机制的细胞移植疗法至关重要，这些策略将对临床前和临床研究产生重大影响。