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

• 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
• 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

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.

缺血性中风是一种主要的医疗负担，对新的治疗方法的需求尚未得到满足 可应用于损伤后狭窄的急性期，在此之后，缺血组织会变成脑梗塞。一个 中风后自然伤口反应的特点是形成包含 非神经细胞核心缺乏特殊的胶质细胞，而神经胶质细胞是支持神经回路和 屏障功能。在斑马鱼和哺乳动物新生儿等具有神经再生能力的生物中， 未成熟的胶质细胞很容易重新填充病变核心，以有效地驱动神经修复，但这种能力已经丧失。 在成年哺乳动物身上。总的目标是促进卒中患者无疤痕的胶质细胞修复以转化病变核心 在允许神经再生的环境中，该项目的主要目标是使用尖端的体内 纵向追踪细胞移植引起的皮质损伤修复结果变化的成像方法 中风，以确定关键的移植参数，导致有效的胶质细胞修复。我们的假设是 活体成像方式可以有效地跟踪移植细胞数量和时间的变化 并量化移植物诱导的微血管密度、灌注量和通透性的变化。 在病变核心。亚急性损伤时移植细胞改变成年中枢神经的性质 系统(CNS)伤口愈合和驱动神经胶质细胞修复代表了一种新的、潜在的变革性策略 这将对中风和其他中枢神经系统损伤的治疗产生广泛影响。在第一个目标中，我们将 纵向追踪神经前体细胞(NPC)和神经前体细胞(NPC)诱导的皮质卒中中神经胶质细胞修复的差异 未成熟的星形胶质细胞移植物。在移植前将移植物植入增殖中的未成熟星形胶质细胞可能会 加快并更好地指导神经胶质细胞的修复过程。我们将使用双光子显微镜(下午2：00)评估移植物 细胞数量、密度、形态和光学相干断层扫描(OCT)对移植诱导的量化 卒中病变核心内及周围血管密度和定量血流的变化。评估回报 对于中枢神经系统屏障功能，我们将使用2 PM对静脉造影剂的渗漏进行量化在第二个 目的：评价卒中后移植时间点对神经胶质细胞修复结果的影响。该项目将提供 细胞移植，包括未成熟的星形胶质细胞移植， 可以通过引导血管生成和修复胶质细胞屏障来重塑病变核心。这项工程将会取得进展 我们对细胞移植的理解，并认证了一个用于评估体内移植功能的工具包，它将 为我们在未来的工作中进一步开展卒中功能恢复研究奠定了基础。