Lung-Brain Crosstalk as a target of cell therapy in ischemic stroke

肺脑串扰作为缺血性中风细胞治疗的目标

基本信息

批准号：
10494236
负责人：
Nikunj B Satani
金额：
$ 19.5万
依托单位：
UNIVERSITY OF TEXAS HLTH SCI CTR HOUSTON
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-09-30 至 2024-02-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10494236
关键词：
Address Adhesions Animals Anti-Inflammatory Agents Applications Grants Binding Blood Circulation Bone Marrow Brain Brain-Derived Neurotrophic Factor Cell Communication Cell Therapy Cells Clinical Research Clinical Trials Coculture Techniques Coupled Culture Media Data Endocrine Endothelial Cells Etiology Experimental Animal Model Exposure to Generations Glucose Goals Health Human Hydrogen Peroxide ICAM1 gene In Vitro Inflammation Inflammatory Interferon Type II Intravenous Ischemia Ischemic Stroke Lead Liver Lung Measures Mediating Mediator of activation protein Messenger RNA Middle Cerebral Artery Occlusion Modeling Mus Neurologic Deficit Neurological outcome Neurons Oxygen Pathway Analysis Pathway interactions Phenotype Plasma Play Primary Cell Cultures Production Protein Tyrosine Kinase RNA Interference Receptor Protein-Tyrosine Kinases Recovery Recovery of Function Research Role Signaling Molecule Site Small Interfering RNA Source Spleen Stimulus Stroke Surgical sutures TNF gene Testing Therapeutic Therapeutic Effect Vascular Endothelial Cell antagonist deprivation design experimental study healing improved in vitro Model in vivo inhibitor ischemic injury mRNA Expression mesenchymal stromal cell neuroprotection neurotrophic factor neutralizing antibody new therapeutic target novel novel therapeutic intervention post stroke promoter receptor regenerative release factor research clinical testing response sham surgery stroke model stroke patient stroke recovery stroke therapy transcriptome sequencing

项目摘要

PROJECT SUMMARY Cell based therapies represent a promising therapeutic approach to enhance stroke recovery. Various research groups have found that bone marrow derived mesenchymal stromal cells (MSCs) improve stroke recovery in experimental animal models. Despite promising results, mechanisms of how MSCs enhance recovery remain unclear. The majority of intravenously (IV) administered MSCs (an approach normally used experimentally and in initial clinical studies), are entrapped in the lungs, while remaining MSCs migrate to liver and spleen, only few cells reach the brain. Studies have shown that while MSCs are short-lived in lungs, their beneficial effects extend for weeks, suggesting that some lung cells are modified by MSCs to deliver endocrine effects that target inflammation and provide trophic responses. Others have tried to improve lung passage of MSCs after IV administration, without considering that this entrapment could in fact be the contributing factor to MSC-mediated benefit. Hence, the main goal of this project is to study the importance of lung-brain crosstalk after IV administration of MSCs in brain recovery. Our preliminary data show that MSCs interact directly with lung endothelial cells (ECs), which increases the release of neurotrophins such as brain-derived neurotrophic factor (BDNF). We also find a robust increase in BDNF mRNA expression in lung ECs upon exposure to MSCs as well as an increase in BDNF levels in plasma from stroke mice after MSC treatment. BDNF exerts its actions through tyrosine receptor kinase B (TrkB). Hence, we hypothesize that MSCs entrapped in lungs prime ECs to release BDNF that acts as a key lung-brain crosstalk mediator, and imparts neuroprotection after stroke by promoting neuronal health and trophicity after stroke. We will pursue the following aims: (1) Using in vitro model of stroke, we will elucidate that lung EC-MSC interaction is the source of BDNF produced in lungs, which benefits brain neuronal integrity. To establish a causal role of lung-released BDNF in neuronal health, we will employ primary lung EC-MSC co-cultures under inflammatory and anti-inflammatory stimulus and perform media transfer experiments on cultured cortical neurons (naïve vs TrkB-deficient), coupled with inhibitor studies using selective TrkB receptor antagonist (ANA-12) and BDNF neutralizing antibody. Specifically to pin down the source of BDNF, we will use RNA-interference (RNAi) studies and silence BDNF in lung EC and/or MSCs to perform co- cultures and media transfer studies. (2) We will determine that “healing” BDNF released from lung ECs after IV MSC treatment in vivo, mediates lung-brain crosstalk and improves functional recovery after stroke. To determine this, we will perform lung specific in vivo silencing of BDNF by intratracheal administration of BDNF siRNA and use these mice to investigate functional recovery from IV MSCs after ischemic stroke. Considering the current limitations in available treatment options for stroke, the advantages of our proposed studies are two- fold: 1) Exploring novel mechanisms (mediated by BDNF) on how MSCs mediate lung-brain crosstalk to promote stroke recovery, and 2) Elucidating a novel targetable pathway that may lead to new treatments for stroke.

项目摘要基于细胞的疗法代表了一种有希望的治疗方法来增强中风恢复。各种研究小组发现，骨髓衍生的间充质基质细胞（MSC）改善了实验动物模型中的中风恢复。尽管结果有希望，但MSC如何增强恢复的机制仍不清楚。大多数静脉内（IV）施用的MSC（通常在实验和最初的临床研究中使用的方法）被插入肺部，而剩余的MSC则迁移到肝脏和脾脏，只有很少的细胞到达大脑。研究表明，尽管MSC在肺部短暂，但其有益作用延长了数周，这表明某些肺部细胞是通过MSC修饰的，以提供靶向注射并提供营养反应的内分泌作用。其他人则试图改善IV施用后MSC的肺部通过，而没有考虑到这种陷阱实际上可能是MSC介导的收益的促成因素。因此，该项目的主要目的是研究静脉注射MSC在大脑恢复中给药后肺脑串扰的重要性。我们的初步数据表明，MSC与肺内皮细胞（ECS）直接相互作用，这增加了神经营养蛋白的释放，例如脑衍生的神经营养因子（BDNF）。我们还发现，在接触MSC时，肺EC中的BDNF mRNA表达有强烈的增加，并且MSC处理后，中风小鼠血浆中血浆中的BDNF水平升高。 BDNF通过酪氨酸受体激酶B（TRKB）发挥作用。因此，我们假设MSC在肺prime EC中释放出充当关键的肺脑串扰介质的BDNF，并通过促进中风后神经元健康和营养性在中风后赋予神经保护作用。我们将追求以下目的：（1）使用中风体外模型，我们将阐明肺EC-MSC相互作用是肺中产生的BDNF的来源，肺部受益于脑神经元完整性。为了在炎症和抗炎刺激下建立肺释放的BDNF在神经元健康中的因果作用，我们将利用培养的皮质神经元（幼稚的与TRKB-DIFIC）对培养的皮质神经元进行培养基转移实验，并使用抑制剂研究中的抑制性Trkniber andf andf andf and-niminf andn-nakogy and-niminf trkbb-niminf and-niminf trkbb-niminf intrk and-niminf trkniby andf。特别是为了固定BDNF的来源，我们将在肺EC和/或MSC中使用RNA - 区分（RNAI）研究（RNAi）研究，并进行静音BDNF来执行共培养和媒体转移研究。（2）我们将确定在体内IV MSC治疗后从肺ECS释放的“愈合” BDNF，介导肺脑串扰并改善中风后的功能恢复。为了确定这一点，我们将通过气管内给药BDNF siRNA对BDNF进行肺特异性体内沉默，并使用这些小鼠研究缺血性中风后从IV MSC中进行功能恢复。考虑到可用治疗方案的当前局限性，我们提出的研究的优势是两倍：1）探索有关MSC如何介导肺脑串扰的新机制（由BDNF介导的）（由BDNF介导）以促进中风恢复，而2）阐明了一种新的目标途径，这些途径可能会导致新的治疗方法。