Meningeal stem cell signals for improving sensorimotor and cognitive outcomes after stroke

脑膜干细胞信号可改善中风后的感觉运动和认知结果

• 批准号: 10087979
• 负责人: Kazuhide Hayakawa
• 金额: $ 39.96万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-02-01 至 2024-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10087979
• 关键词: AMD3100; Accounting; Animal Model; Astrocytes; BMP2 gene; Behavior assessment; Blood - brain barrier anatomy; Blood Vessels; Brain; Brain Injuries; Brain-Derived Neurotrophic Factor; CCL3 gene; CD 200; CRISPR/Cas technology; CXCR4 gene; Cell Culture Techniques; Cell Death; Cells; Cerebral Ischemia; Cerebral cortex; Coculture Techniques; Cognitive; Data; Defect; Development; Endothelium; Female; Genes; Goals; Growth Factor; HDAC3 gene; Human; Infarction; Injury; Ischemia; Label; Literature; Location; Maps; Mediating; Mediator of activation protein; Membrane; Meningeal; Meninges; Methods; Modeling; Modification; Molecular; Multipotent Stem Cells; Mus; Neuroglia; Neuronal Injury; Neurons; Outcome; Pathway interactions; Patients; Pericytes; Pharmacology; Process; Proliferating; Publishing; Radial; Rattus; Recovery; Signal Transduction; Source; Stroke; Stromal Cell-Derived Factor 1; Testing; Therapeutic; Transforming Growth Factor beta; Up-Regulation; Western Blotting; activin A; base; behavior test; blood damage; brain endothelial cell; cognitive recovery; experimental study; functional outcomes; improved; in vivo; in vivo Model; in vivo imaging; inhibitor/antagonist; loss of function; macrophage; male; migration; mortality; mouse model; neurogenesis; neurological recovery; neurovascular; neurovascular unit; neutralizing antibody; new therapeutic target; novel; novel strategies; optical imaging; post stroke; post stroke dementia; receptor; recombinase-mediated cassette exchange; relating to nervous system; repaired; response; restoration; spatiotemporal; stem cell migration; stem cell niche; stem cell proliferation; stem cells; stroke recovery; therapeutic target; tool

Meningeal stem cell signals for improving sensorimotor and cognitive outcomes after stroke Mortality after stroke has decreased. The challenge now is how to deal with the accumulating burden of sensorimotor deficits and post-stroke dementia in surviving patients. In this proposal, we will investigate the meninges as a novel source of stem cell signals that may potentially assist with neurovascular unit recovery. Our pilot data suggest that: (i) meningeal multipotent stem cells (MeSCs) migrate to perivascular space after focal ischemia, (ii) MeSC response may be significant, accounting for up to 30% of CD271 “total stem cell response”, (iii) this is an “active process” since inhibiting CXCR4 decreased MeSC migration, (iv) MeSC response may be a relevant therapeutic target since it can be significantly amplified by blocking CD271- mediated stem cell death with LM11A-31, whereas blocking SDF-1 signaling with AMD3100 decreased MeSC migration and worsened blood-brain barrier (BBB) injury; (v) beneficial actions of MeSC may involve restoration of vascular integrity and upregulation of TGF-β-stimulated macrophages that further promote neurogenic responses, and finally (vi) models and methods to dissect this phenomenon are feasible including molecular tools, vasculome mapping, behavioral tests, and in vivo imaging in our collaborating labs. Based on our pilot data, we hypothesize that (a) brain endothelial cells and astrocytes secrete SDF-1 into perivascular space to attract MeSCs, (b) MeSCs restore damaged BBB and rescue the vasculome, (c) MeSC crosstalk with perivascular macrophages promotes TGF-β-mediated neurogenesis, and (d) MeSC-mediated neurovascular unit recovery ameliorates sensorimotor deficits and post-stroke dementia. We have 3 specific aims. In Aim 1, we will map the profile of MeSC proliferation, accumulation, and differentiation in focal cerebral ischemia. In Aim 2, we will investigate cellular mechanisms of crosstalk between MeSCs and vascular/perivascular cells that underlie MeSC migration, BBB repair, vasculome renormalization and neurogenesis. In Aim 3, we will use gain and loss-of-function experiments to modify MeSC-mediated neurovascular remodeling, and assess sensorimotor and cognitive outcomes in vivo. Our experiments will utilize cell cultures and animal models. Molecular tools include a combination of pharmacologic activators/inhibitors, CRISPR/cas9, and cre-lox systems to modify specific pathways and genes. Translational relevance will be assessed with in vivo imaging and long-term outcomes. This project should define a novel mechanism wherein meningeal stem cells communicate with the neurovascular unit, and hopefully provide new approaches to improve sensorimotor recovery and ameliorate post-stroke dementia.

脑卒中后脑膜干细胞信号改善感觉运动和认知结果 中风后死亡率下降。现在的挑战是如何处理日益加重的 存活患者的感觉运动缺陷和卒中后痴呆。在本提案中，我们将调查 脑膜作为干细胞信号的新来源，可能有助于神经血管单位的恢复。 我们的初步数据表明：（i）脑膜多能干细胞（MeSC）在移植后迁移到血管周围空间， （ii）MeSC反应可能是显著的，占CD 271-总干细胞的高达30%， （iii）这是一个“主动过程”，因为抑制CXCR 4降低了MeSC的迁移，（iv）MeSC 反应可能是相关的治疗靶点，因为它可以通过阻断CD 271- 用LM 11 A-31介导干细胞死亡，而用AMD 3100阻断SDF-1信号传导降低MeSC 迁移和恶化的血脑屏障（BBB）损伤;（v）MeSC的有益作用可能涉及 血管完整性的恢复和TGF-β刺激的巨噬细胞的上调， 神经性反应，最后（vi）剖析这种现象的模型和方法是可行的，包括 分子工具、血管组绘图、行为测试和我们合作实验室的体内成像。 基于我们的初步数据，我们假设（a）脑内皮细胞和星形胶质细胞分泌SDF-1进入脑内， 血管周围空间以吸引MeSC，（B）MeSC恢复受损的BB B并拯救血管组，（c）MeSC 与血管周围巨噬细胞的串扰促进TGF-β介导的神经发生，和（d）MeSC介导的神经发生。 神经血管单位恢复改善感觉运动缺陷和中风后痴呆。 我们有三个具体目标。在目标1中，我们将绘制MeSC增殖、积累和增殖的分布图。 局灶性脑缺血的分化。在目标2中，我们将研究细胞之间的串扰机制， MeSC和血管/血管周围细胞是MeSC迁移、BBB修复、血管组重整的基础 和神经发生。在目标3中，我们将使用功能获得和丧失实验来修饰MeSC介导的 神经血管重塑，并评估体内感觉运动和认知结果。我们的实验将 利用细胞培养和动物模型。分子工具包括药理学的组合， 激活剂/抑制剂、CRISPR/cas9和cre-lox系统来修饰特定的途径和基因。平移 相关性将通过体内成像和长期结果进行评估。这个项目应该定义一部小说 脑膜干细胞与神经血管单位沟通的机制，并有望提供新的 改善感觉运动恢复和改善中风后痴呆的方法。