Multimodal approach investigating the immunomodulatory effect ofneural stem cells in stroke recovery

人类神经干细胞(HNSC)治疗中风的前景看好。 进行早期临床试验，以治疗中风引起的长期残疾。这 为数百万患有中风的慢性衰弱影响的美国人带来了希望。 然而，主要的问题仍然是干细胞注射到大脑中如何驱动中风 恢复。干细胞作用机制的一个线索是最近发现的阳性 卒中患者脑部MRI信号T2-FLAIR信号与卒中恢复的相关性 接受干细胞治疗的患者。我们已经成功地复制了这种干细胞诱导的天赋 在中风损伤大鼠中的信号，并表明它与炎症有关。这导致了我们的 中心假设：干细胞移植通过诱导再生来推动康复 炎症反应。这项资助的目的是使用皮质下中风的大鼠模型来 探讨卒中慢性期移植人神经干细胞的免疫调节作用。 在区域、细胞和分子层面使用多模式方法。在Aim1中，我们使用MRI 和PET成像，以确定hNSC后哪些脑区显示炎症变化 移植，以及哪些炎症区与康复最相关。这也将考验 这些临床相关的成像方式作为中风恢复的生物标记物的效用。在……里面 目的：我们使用多种工具识别hNSC治疗引起的免疫变化。 描述存在的免疫细胞的类型和分子特征，以及它们的空间 互动。在目标3中，我们确定了哪些hNSC分泌的因子调节免疫。 通过使用CRISPR工具在体外和体内测试几个候选基因来调节反应 关键候选因素的表达。我们将研究操纵这些水平的影响 蛋白质对中风康复和免疫调节的影响。研究完成后，我们将拥有 在理解hNSC诱导的免疫调节如何影响方面取得了重大进展 大脑修复。这一贡献是重大的，因为它将：a)识别潜在的生物标记物，两者 治疗前后，对于hNSC诱导的恢复；b)开始描绘分子 参与脑修复的途径；以及c)最终导致确定新的治疗方法 卒中。

Human neural stem cell (hNSC) therapy for stroke is showing promise as it moves from the bench into early clinical trials to treat the long term disabilities resulting from stroke. This provides hope for the millions of Americans living with the chronic, debilitating effects of stroke. Major questions remain, however, about how stem cells injected into the brain drive stroke recovery. A clue to stem cell mechanism of action is the recent discovery of the positive correlation between stroke recovery and a brain MRI signal – T2-FLAIR signal– in stroke patients treated with stem cells. We have successfully reproduced this stem cell-induced FLAIR signal in stroke-injured rats, and shown that its associated with inflammation. This led to our central hypothesis: stem cell transplantation drives recovery by inducing a regenerative inflammatory response. The objective of this grant is to use a rat model of subcortical stroke to investigate the immunomodulatory effects of hNSC transplanted at the chronic stage of stroke, at the regional, cellular and molecular levels using a multimodal approach. In Aim1 we use MRI and PET imaging to identify which brain regions show inflammatory changes after hNSC transplantation, and which inflammatory regions best correlate with recovery. This will also test the utility of these clinically relevant imaging modalities as biomarkers for stroke recovery. In Aim 2 we identify the immune changes induced by hNSC treatment using multiple tools to characterize the types and molecular signatures of the immune cells present, and their spatial interactions. In Aim 3 we determine which hNSC-secreted factors modulate the immune response by testing several candidates, in vitro and in vivo, using CRISPR tools to modulate expression of key candidate factors. We will study the impact of manipulating the levels of these proteins on stroke recovery and immunomodulation. Upon conclusion of the study, we will have made significant advancements in understanding how hNSC-induced immunomodulation affects brain repair. This contribution is significant because it will: a) identify potential biomarkers, both pre- and post-treatment, for hNSC-induced recovery; b) begin to delineate the molecular pathways involved in brain repair; and c) ultimately lead to identification of novel therapies for stroke.