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

脑膜干细胞信号对改善中风后感觉运动和认知结局的作用 中风后的死亡率已经下降。现在的挑战是如何应对不断积累的负担 存活患者的感觉运动缺陷和卒中后痴呆。在这项建议中，我们将调查 脑膜作为一种新的干细胞信号来源，可能潜在地帮助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(MeSCs)在 局灶性缺血，(Ii)MESC反应可能显著，占CD271“干细胞总数的30% 反应“，(三)这是一个”积极的过程“，因为抑制CXCR4减少了MeSC的迁移，(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- LM11A-31介导干细胞死亡，AMD3100阻断SDF-1信号减少MeSC 迁移和加重血脑屏障(BBB)损伤；(V)MeSC的有益作用可能包括 血管完整性的恢复和转化生长因子-β刺激的巨噬细胞上调进一步促进 神经源性反应，最后(Vi)分析这种现象的模型和方法是可行的，包括 分子工具、血管组图谱、行为测试和活体成像。 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SDF-1分泌到 血管周围空间以吸引MeSCs，(B)MeSCs恢复受损的血脑屏障并拯救血管小体，(C)MeSC 与血管周围巨噬细胞的串扰促进转化生长因子-β介导的神经发生，以及(D)MeSC介导的 神经血管单位恢复可改善感觉运动障碍和卒中后痴呆。 我们有三个具体目标。在目标1中，我们将绘制MeSC的增殖、积累和 局灶性脑缺血的鉴别诊断。在目标2中，我们将研究串扰的细胞机制 MESC和作为MESC迁移、血脑屏障修复、血管小体重整化基础的血管/周围细胞 和神经发生。在目标3中，我们将使用获得和功能损失实验来修饰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/抑制物、CRISPR/Cas9和cre-lox系统，以修饰特定的途径和基因。翻译 相关性将通过活体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 促进感觉运动恢复和改善卒中后痴呆的方法。

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.