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

基本信息

批准号：
10528477
负责人：
Kazuhide Hayakawa
金额：
$ 39.21万
依托单位：
MASSACHUSETTS GENERAL HOSPITAL
依托单位国家：
美国
项目类别：
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/10528477
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 CD 200 CRISPR/Cas technology CXCR CXCR4 gene Cell Communication Cell Culture Techniques Cell Death Cells Cerebral Ischemia Cerebral cortex Coculture Techniques Cognitive Collaborations Cre lox recombination system Data Defect Development Endothelium Female Genes Goals Growth Factor HDAC3 gene Human Infarction Injury Ischemia Label Literature Location Macrophage Maps Mediating Mediator Membrane Meningeal Meninges Methods Modeling Modification Molecular Multipotent Stem Cells Mus Neuroglia Neuronal Injury Neurons Outcome Pathway interactions Patients Pericytes Process 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 behavior test blood damage brain endothelial cell cognitive recovery endothelial repair experimental study functional outcomes gain of function improved in vivo in vivo Model in vivo imaging inhibitor loss of function male migration mortality mouse model neural neurogenesis neurological recovery neurovascular neurovascular unit neutralizing antibody new therapeutic target novel novel strategies optical imaging pharmacologic post stroke post stroke dementia receptor repaired response restoration spatiotemporal stem cell migration stem cell niche stem cell proliferation stem cells stroke cognitive outcome stroke recovery therapeutic target tool

项目摘要

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.

脑膜干细胞信号可改善中风后的感觉运动和结果中风后的死亡率有所下降，现在的挑战是如何应对不断累积的负担。在本提案中，我们将调查幸存患者的感觉运动缺陷和中风后痴呆。脑膜作为干细胞信号的新来源，可能有助于神经血管单元的恢复。我们的试验数据表明：(i) 脑膜多能干细胞 (MeSC) 迁移至血管周围空间局灶性缺血，(ii) MeSC 反应可能很显着，占 CD271“总干细胞的 30%” 响应”，(iii) 这是一个“主动过程”，因为抑制 CXCR4 会减少 MeSC 迁移，(iv) MeSC 反应可能是一个相关的治疗靶点，因为它可以通过阻断 CD271- 来显着放大。用 LM11A-31 介导干细胞死亡，而用 AMD3100 阻断 SDF-1 信号传导可减少 MeSC 迁移和加剧血脑屏障（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，进一步促进神经源性反应，最后（vi）剖析这种现象的模型和方法是可行的，包括我们合作实验室的分子工具、血管组图谱、行为测试和体内成像。根据我们的试验数据，我们发现 (a) 脑内皮细胞和星形胶质细胞将 SDF-1 分泌到血管周围空间吸引 MeSC，(b) MeSC 恢复受损的 BBB 并拯救血管，(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，以及 (d) MeSC 介导神经血管单位恢复可改善感觉运动缺陷和中风后痴呆。我们有 3 个具体目标，在目标 1 中，我们将绘制 MeSC 增殖、积累和分布的概况。在目标 2 中，我们将研究细胞之间串扰的机制。 MeSC 和血管/血管周围细胞是 MeSC 迁移、BBB 修复、血管正常化的基础在目标 3 中，我们将使用功能获得和丧失实验来修改 MeSC 介导的功能。我们的实验将进行神经血管重塑，并评估体内感觉运动和认知结果。利用细胞培养物和动物模型，包括药理学的组合。激活剂/抑制剂、CRISPR/cas9 和 cre-lox 系统来修改特定的途径和基因。该项目将通过体内成像和长期结果来评估相关性。机制提供脑膜干细胞与神经血管单元的沟通，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。