Intravital imaging of transplant evoked glia repair in stroke

移植诱发中风神经胶质修复的活体成像

基本信息

批准号：
10741583
负责人：
Timothy Mark O'Shea
金额：
$ 24.75万
依托单位：
BOSTON UNIVERSITY (CHARLES RIVER CAMPUS)
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-07-01 至 2025-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10741583
关键词：
Acceleration Acute Address Adult Astrocytes Axon Behavioral Blood Vessels Blood flow Cell Count Cell Transplantation Cells Central Nervous System Certification Cicatrix Clinical Cognitive deficits Competence Contrast Media Cues Effectiveness Environment Extravasation Fibrosis Fostering Future Goals Growth Healthcare Hour Immune Immunohistochemistry Infarction Infiltration Injury Intravenous Ischemia Ischemic Stroke Knowledge Lesion Mammals Maps Methods Modeling Molecular Morphology Multimodal Imaging Mus Natural regeneration Nature Nerve Regeneration Neuroglia Neurons Neurophysiology - biologic function Optical Coherence Tomography Organism Outcome Perfusion Peripheral Permeability Phase Phenotype Population Positioning Attribute Process Proliferating Recovery Recovery of Function Reporter Saline Serum Proteins Source Stroke System Time Tissues Translations Transplantation Work Zebrafish angiogenesis care burden central nervous system injury cohort density imaging modality improved in vivo insight intravital imaging multimodality neonate nerve stem cell neural circuit neural repair neurovascular novel permissiveness post stroke repair function repaired restoration stroke therapy two photon microscopy wound healing wound response

项目摘要

Ischemic stroke is a major healthcare burden and there is an important unmet need for new treatments that can be applied beyond the narrow acute phase of injury after which ischemic tissue becomes infarcted. A hallmark of natural wound responses after stroke is the formation of compartmentalized lesions that contain non-neural cell cores that lack specialized glia cells that are fundamental to supporting neural circuits and barrier functions. In organisms capable of neural regeneration such as zebrafish and the mammalian neonate, immature glia cells readily repopulate lesion cores to effectively drive neural repair, but this competency is lost in adult mammals. With the overall goal of promoting scar-free glia repair in stroke to transform lesion cores into neural regeneration permissive environments, this project’s main objective is to use cutting-edge intravital imaging methods to longitudinally track cell graft evoked changes in wound repair outcomes following cortical strokes to identify critical graft parameters that lead to effective glia-repair. Our hypothesis is that multiple intravital imaging modalities can be used to effectively track temporal changes in grafted cell number and phenotype as well as quantify graft induced alterations to microvasculature density, perfusion, and permeability in lesion cores. Grafting cells during the sub-acute injury phase to alter the nature of adult central nervous system (CNS) wound healing and drive glia repair represents a novel and potentially transformative strategy that would have broad impact for treatment of stroke and other CNS injuries. In the first aim, we will longitudinally track differences in glia repair in cortical strokes directed by neural progenitor cells (NPC) and immature astroglia grafts. Priming grafts into proliferating immature astrocytes prior to transplantation may accelerate and better guide glia repair processes. Using two-photon microscopy (2PM) we will evaluate graft cell number, density, and morphology and optical coherence tomography (OCT) to quantify graft induced changes in vascular density and quantitative blood flow in and around stroke lesion cores. To evaluate return of CNS barrier functions we will quantify the leakage of intravenous contrast agents using 2PM. In the second aim, we evaluate the effect of post stroke grafting timepoint on glia repair outcomes. This project will provide mechanistically validated, proof-of-principle evidence into how cell grafts, including immature astrocyte grafts, can remodel lesion cores by directing angiogenesis and restoration of glia barriers. This project will advance our understanding of cell transplantation and certify a toolkit for evaluating transplant functions in vivo that will position us to pursue advanced stroke functional recovery studies in future work.

缺血性中风是伯宁的主要医疗保健，对新疗法的重要需求很重要可以超出狭窄的损伤急性阶段的应用，此后缺血组织被梗塞。一个中风后自然伤口反应的标志是形成了包含的隔室性病变缺乏专门的神经胶质细胞的非神经细胞核，这些细胞是支持神经回路和障碍功能。在能够进行神经再生的生物中，例如斑马鱼和哺乳动物新生儿，未成熟的神经胶质细胞容易复制病变核心以有效驱动神经修复，但这种能力丧失在成年哺乳动物中。总体目的是促进中风中无疤痕的维修以改变病变核心进入神经代理的允许环境，该项目的主要目标是使用尖端的插入纵向跟踪细胞移植的成像方法诱发了皮质后伤口修复结果的变化中风以识别导致有效神经胶质修复的关键移植参数。我们的假设是多个插入成像方式可用于有效跟踪移植细胞数和表型以及量化移植物引起的微脉管密度，灌注和渗透率的改变在病变核心。在亚急性损伤阶段嫁接细胞以改变成人中枢神经的性质系统（CNS）伤口愈合和驱动神经胶质修复代表了一种新颖而潜在的变革策略这将对中风和其他中枢神经系统损伤的治疗产生广泛的影响。在第一个目标中，我们将纵向跟踪神经祖细胞（NPC）和未成熟的星形胶质细胞移植物。在移植之前将移植物启动到增殖的未成熟星形胶质细胞中加速和更好的指导胶质修复过程。使用两光子显微镜（下午2点），我们将评估移植物细胞数，密度和形态和光学相干断层扫描（OCT），以量化移植物诱导的中风病变核心内外的血管密度和定量血流的变化。评估回报 CNS屏障功能的函数我们将使用2pm量化静脉造影剂的泄漏。在第二个目的，我们评估了中风后移植时间点对胶质修复结果的影响。这个项目将提供通过机械验证的原则证据证明了细胞移植的方式，包括未成熟的星形胶质细胞移植物，可以通过指导血管生成和恢复神经胶质屏障的重塑病变核心。这个项目将进步我们对细胞移植的理解并认证用于评估体内移植功能的工具包将我们定位为在未来的工作中追求先进的中风功能恢复研究。