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：
10307121
负责人：
Kazuhide Hayakawa
金额：
$ 39.59万
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/10307121
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 Cre lox recombination system 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 endothelial repair experimental study functional outcomes improved in vivo in vivo Model in vivo imaging inhibitor 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 relating to nervous system repaired response restoration spatiotemporal stem cell migration stem cell niche stem cell proliferation stem cells 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，用于改善中风后的感觉运动和认知结果中风后的死亡率恶化。现在的挑战是如何应对累积的燃烧感觉运动定义和震后痴呆症中的生存患者。在此提案中，我们将调查脑膜是一种新型的干细胞信号来源，可能有助于神经血管单位恢复。我们的试点数据表明：（i）脑膜多能干细胞（MESC）在局灶性缺血，（ii）MESC响应可能很明显，最多占CD271的30％“干细胞” 响应”，（iii）这是一个“主动过程”，因为抑制CXCR4降低了MESC迁移，（iv）MESC 反应可能是相关的治疗靶标，因为它可以通过阻止CD271--可以显着放大。 LM11A-31介导的干细胞死亡，而AMD3100的SDF-1信号传导降低了MESC 迁移和恶化的血脑屏障（BBB）损伤；（v）MESC的有益行动可能涉及恢复血管完整性和TGF-β刺激的巨噬细胞的上调，这些巨噬细胞进一步促进神经源性反应，最后是（vi）模型和剖析这种现象的方法，包括分子工具，脉管映射，行为测试和我们的协作实验室中的体内成像。根据我们的试验数据，我们假设（a）脑内皮细胞和星形胶质细胞将Secret SDF-1纳入血管周空间吸引MESC，（b）MESC恢复受损的BBB并营救Vasculome，（c）MESC 与周围巨噬细胞的串扰可促进TGF-β介导的神经发生，并且（d）MESC介导神经血管单位恢复可以改善感觉运动定义和触诊后痴呆症。我们有3个具体目标。在AIM 1中，我们将绘制MESC增殖，积累和局灶性脑缺血的分化。在AIM 2中，我们将研究串扰的细胞机制 MESC和血管/血管细胞，其基于MESC迁移，BBB修复，血管重新归一化的基础和神经发生。在AIM 3中，我们将使用获得和功能丧失实验来修改MESC介导的神经血管重塑，并在体内评估感觉运动和认知结果。我们的实验会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系统可修改特定的途径和基因。翻译相关性将通过体内成像和长期结局进行评估。这个项目应该定义小说脑膜干细胞与神经血管单元进行通信的机制，并希望提供新的改善感觉运动恢复和改善势后痴呆症的方法。