Adrenergic receptor modulation of MSC exosome cargo to improve wound healing

MSC 外泌体货物的肾上腺素受体调节可改善伤口愈合

基本信息

批准号：
10112831
负责人：
Roslyn Rivkah ISSEROFF
金额：
$ 16.75万
依托单位：
UNIVERSITY OF CALIFORNIA AT DAVIS
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：
2020-03-01 至 2022-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10112831
关键词：
Adrenergic Receptor Affect Agonist American Animal Model Anti-Inflammatory Agents Apoptosis Attention Bone Marrow Catecholamines Cells Chronic Clinical Complement Cytokine Signaling Data Development Environment Epinephrine Etiology Foundations Future Generations Goals Growth Factor Health Health Expenditures Hormones Impaired healing Impaired wound healing In Vitro Infection Inflammation Inflammatory Inflammatory Response Ligands Lipids Mesenchymal Mesenchymal Stem Cells MicroRNAs Neuroimmunomodulation Norepinephrine Parents Patients Phenotype Process Property Proteomics Public Health Quality of life Role Signal Transduction Site Stress System T-Cell Proliferation Therapeutic Therapeutic Agents Timolol Varicose Ulcer Work Wound models beta-2 Adrenergic Receptors chronic ulcer chronic wound cytokine db/db mouse decubitus ulcer diabetic ulcer effective therapy exosome healing immunoregulation improved in vivo interest paracrine receptor binding regenerative repair function standard care stem cell exosomes stem cells therapeutic candidate transcriptomics wound wound bed wound environment wound healing wound treatment

项目摘要

Chronic wounds, defined as those that do not progress through the normal healing process and remain unhealed after one month of standard care, pose tremendous health problems- clinically and economically. There is growing interest in isolating ttherapeutic agents directly from mesenchymal stem cells: particularly the exosome enriched fractions that are now understood to carry the active immunomodulatory and regenerative factors. Their cargo of miRNAs, Wnt ligands, growth factors, cytokines and signaling lipids provide the mechanism for paracrine stimulation of wound resident cells. However, cargo contents of MEX are dependent on culture conditions. MSCs express a full complement of AR, including the β2-AR. Our prior work has demonstrated that β2-AR agonists increase inflammatory cytokine secretion MSCs and conversely, treating MSC with a β2-AR antagonist, timolol, substantially enhances their anti-inflammatory and wound reparative properties. Here we hypothesize that activation of MSC β2-AR by stress catecholamine agonists alters their exosomal cargo contents, to be more pro-inflammatory, and conversely, β2-AR antagonists block agonist- induced changes, and revert exosomal cargo to pro-reparative, anti-inflammatory phenotype. Our long-term goal is to determine whether priming MSC with a β2-AR antagonist can generate MSC exosomes (MEX) that are a viable therapeutic candidate for improving wound healing. Our major objective is to evaluate the effects of β2-AR activation and antagonism on MEX cargo contents and function in improving healing. In Specific Aim 1a: we will valuate the effects of activation of the β2-AR on MEX cargo. Determine if stress catecholamine ligands modify the cargo to become more pro-inflammatory. Proteomic, and transcriptomic analyses of MEX contents will be performed. Their pro-inflammatory potential will be evaluated by cytokine release and in vitro T cell proliferation. In Aim 1b we will evaluate the effects of blockade of the β2-AR on MEX cargo. The endogenous generation of catecholamines by MSC will be determined, and the effect of blockade of their activation of MSC on exosome cargo will be analyzed as in Aim 1a. In Specific Aim 2: we will determine whether β2-AR agonists and antagonists alternatively result in MEX with diminished or enhanced wound healing properties, respectively, in an in vivo impaired healing wound model (db/db mouse). This work will provide foundational information for the future development of MEX as a wound therapeutic.

慢性伤口，定义为未在正常愈合过程中进展并保持的慢性伤口经过一个月的标准护理，在临床和经济上构成了巨大的健康问题。直接从间充质干细胞中分离甲状腺素剂的兴趣越来越大：尤其是外泌体富集的馏分，这些馏分现在已经理解为携带主动免疫调节和再生因素。它们的miRNA货物，Wnt配体，生长因子，细胞因子和信号传导脂质提供了伤口居民细胞的旁分泌刺激机制。但是，MEX的货物是依赖的关于文化条件。 MSC表达了包括β2-AR在内的AR完整完成。我们先前的工作有证明β2-ar激动剂增加了炎性细胞因子分泌MSC，相反，治疗 MSC具有β2-AR拮抗剂Timolol，大大增强了其抗炎和伤口的修复特性。在这里，我们假设应力儿茶酚胺激动剂对MSCβ2-AR的激活改变了它们外泌体货物含量更为促炎，相反，beta2-ar拮抗剂阻止了激动剂 - 诱导的变化，并将外泌体货物恢复为依赖性的抗炎表型。我们的长期目标是确定用β2AR拮抗剂启动MSC是否可以产生MSC外泌体（MEX）是改善伤口愈合的可行治疗候选者。我们的主要目标是评估效果 β2-ar激活和对MEX货物含量的拮抗作用以及改善愈合的功能。具体 AIM 1A：我们将重视β2-AR激活对MEX货物的影响。确定压力儿茶酚胺是否配体修改货物以变得更加促炎。 MEX的蛋白质组学和转录组分析将执行内容。它们的促炎潜力将通过细胞因子释放和体外评估细胞增殖。在AIM 1B中，我们将评估β2-AR封锁对MEX货物的影响。这将确定MSC的内源性儿茶酚胺的生成，并将其阻断的影响在AIM 1A中将分析MSC在外泌体货物上的激活。在特定目标2中：我们将确定 β2-ar激动剂和拮抗剂是否会导致MEX减少或增强在体内损害伤口模型（DB/DB小鼠）中，分别愈合特性。这项工作将为MEX作为伤口疗法的未来发展提供基本信息。