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

多模式方法研究神经干细胞在中风恢复中的免疫调节作用

• 批准号: 10331043
• 负责人: GARY K STEINBERG
• 金额: $ 60.98万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-08-01 至 2024-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10331043
• 关键词: Address; Affect; Aftercare; American; Animal Model; Area; Astrocytes; Biological Markers; Biological Process; Brain; Brain Injuries; Brain region; CRISPR/Cas technology; Cell Therapy; Cell Transplantation; Cell model; Cells; Chronic; Clinical; Clinical Data; Clinical Trials; Coculture Techniques; Control Groups; Coupled; Data; Distant; Encephalitis; Exhibits; Family; Female; Flow Cytometry; Genes; Goals; Grant; Human; Image; Immune; Immune response; Immunohistochemistry; In Vitro; Inflammation; Inflammatory; Inflammatory Response; Injections; Injury; Lead; Magnetic Resonance Imaging; Microglia; Modeling; Molecular; Molecular Profiling; Monitor; Myelogenous; Needles; Ontology; Pathway interactions; Patients; Peripheral; Positron-Emission Tomography; Proteins; Rattus; Recovery; Recovery of Function; Reporting; Rodent; Running; Signal Transduction; Site; Stem cell transplant; Stroke; System; Techniques; Testing; Therapeutic; Therapeutic Effect; Time; Tracer; Transplantation; brain magnetic resonance imaging; brain repair; cell type; clinical imaging; clinically relevant; disability; early phase clinical trial; imaging modality; immune activation; immunoregulation; improved; in vivo; injured; interest; macrophage; male; multimodality; nerve stem cell; new therapeutic target; novel; novel therapeutics; patient population; post stroke; potential biomarker; regenerative; side effect; single-cell RNA sequencing; societal costs; stem cell therapy; stem cells; stroke model; stroke patient; stroke recovery; stroke therapy; tool; transcriptome

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.

人类神经干细胞 (hNSC) 治疗中风的前景随着它从 进入早期临床试验以治疗中风引起的长期残疾。这 为数百万遭受中风慢性、衰弱影响的美国人带来了希望。 然而，关于注射到大脑中的干细胞如何驱动中风仍然存在主要问题 恢复。干细胞作用机制的一个线索是最近发现的积极的 中风恢复与脑部 MRI 信号（T2-FLAIR 信号）之间的相关性 接受干细胞治疗的患者。我们已经成功复制了这种干细胞诱导的 FLAIR 中风损伤大鼠的信号，并表明其与炎症有关。这导致了我们的 中心假设：干细胞移植通过诱导再生来促进恢复 炎症反应。该拨款的目的是利用皮质下中风的大鼠模型来 研究中风慢性期移植的 hNSC 的免疫调节作用， 使用多模式方法在区域、细胞和分子水平上进行研究。在 Aim1 中，我们使用 MRI 和 PET 成像以确定 hNSC 后哪些大脑区域显示炎症变化 移植，以及哪些炎症区域与恢复最相关。这也将测试 这些临床相关成像方式作为中风恢复生物标志物的用途。在 目标 2 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，我们确定哪些 hNSC 分泌因子调节免疫 通过使用 CRISPR 工具在体外和体内测试几种候选物来调节反应 关键候选因素的表达。我们将研究操纵这些水平的影响 中风恢复和免疫调节的蛋白质。研究结束后，我们将得到 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。这一贡献意义重大，因为它将：a) 识别潜在的生物标志物， 治疗前和治疗后，用于 hNSC 诱导的恢复； b) 开始描绘分子 参与大脑修复的途径； c）最终导致新疗法的确定 中风。