Human Mesenchymal Stem Cell Microvesicles for the Treatment of Acute Lung Injury

人间充质干细胞微泡治疗急性肺损伤

• 批准号: 9173912
• 负责人: Jae Woo Lee
• 金额: $ 56.57万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-05-01 至 2021-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9173912
• 关键词: ABCC1 gene; ATP-Binding Cassette Transporters; Acute Lung Injury; Adjuvant; Adult Respiratory Distress Syndrome; Affect; Agonist; Alveolar; Alveolar Macrophages; Alveolus; Animal Model; Anti-Inflammatory Agents; Anti-inflammatory; Antibiotics; Bacteria; Bacterial Counts; Bacterial Pneumonia; Biological; Blood Circulation; Blood Vessels; Bone Marrow; Cell Communication; Cell membrane; Cells; Chloride Channels; Clinical Trials; Cystic Fibrosis Transmembrane Conductance Regulator; Endothelium; Epithelial; Escherichia coli; Glycocalyx; Goals; Grant; Growth; Growth Factor; Home environment; Human; Iatrogenesis; Impairment; Inflammation; Inflammatory; Innovative Therapy; Leukotriene A4; Leukotriene B4; Leukotrienes; Lipopolysaccharides; Liquid substance; Lung; Malignant - descriptor; Mediating; Membrane; Mesenchymal Stem Cells; Messenger RNA; MicroRNAs; Modeling; Multidrug Resistance-Associated Proteins; Mus; Nitric Oxide; Organelles; Parents; Patients; Peptide Receptor; Permeability; Pharmacological Treatment; Phenotype; Pneumonia; Poly I-C; Pre-Clinical Model; Property; Proteins; Pseudomonas; Pseudomonas aeruginosa pneumonia; Pulmonary Edema; Pulmonary artery structure; Reporting; Research Personnel; Resistance; Risk; Role; Safety; Sepsis; Sheep; Site; Sodium Channel; Stem cells; Surface; Syndrome; TLR3 gene; Testing; Therapeutic; Therapeutic Effect; Tissues; Translations; Type II Epithelial Receptor Cell; Vesicle; antimicrobial; antimicrobial peptide; apical membrane; cell type; cytokine; epithelial Na+ channel; exosome; experimental study; hemodynamics; improved; injured; killings; lipid metabolism; lung injury; macrophage; microvesicles; monocyte; mortality; mouse model; neutrophil; novel; paracrine; preclinical study; preconditioning; pressure; prevent; smoke inhalation; tissue repair; tumor; tumor growth

PROJECT SUMMARY/ABSTRACT Acute respiratory distress syndrome remains a devastating syndrome affecting more than 200,000 patients annually in the U.S. with a mortality rate approaching 40%. Currently, there are no pharmacological treatments available that reduce mortality. Consequently, innovative therapies are needed. In various preclinical models of acute lung injury, mesenchymal stem cells (MSC) have been shown to secrete multiple paracrine factors that can reduce lung endothelial and epithelial permeability, decrease inflammation, enhance tissue repair, and inhibit bacterial growth, ultimately decreasing mortality. Despite a favorable safety profile in early clinical trials, however, MSC have the capacity for spontaneous malignant transformation following multiple passages in culture as well as the ability to promote tumor growth in small animal models. In the prior grant period, we found that microvesicles (MV) released by human MSCs were as biologically active as the parent stem cells in both inflammatory and infectious causes of acute lung injury. Once considered cellular debris, MVs, anuclear circular membrane bound vesicles 50 nm to 200 nm in size that are constitutively released by many cell types, are now recognized as important mechanisms for cell-cell communication. Our overall hypothesis for the current proposal is that MSC MVs are biologically active, and its therapeutic activity in severe bacterial pneumonia is mediated through transfer of mRNAs, microRNAs, proteins and/or organelles from the MVs to the injured alveolus. In Aim 1, we will further uncover the therapeutic mechanisms by studying the effect of human MSC MVs on lipid metabolism and the activity of ATP-binding cassette transporter, multidrug resistance associated protein (MRP)1, in mice injured with severe pneumonia. We hypothesize that MSC MVs will inhibit MRP1 activity in monocytes/macrophages and decrease the enzymatic conversion and secretion of leukotriene (LT)A4 to LTC/D/E4, leukotrienes involved in lung protein permeability, and increase the conversion into LTB4, leukotriene involved in bacterial killing. In Aim 2, we will test the effect of human MSC MV on net fluid transport in both a novel ex vivo perfused human lung injured with severe bacterial pneumonia and in primary cultures of human alveolar epithelial type II cells injured by an inflammatory insult. We hypothesize that MSC MVs will prevent pulmonary edema formation by restoring the apical membrane expression of the major epithelial sodium channel, αENaC. In Aim 3, we will determine if human MSC MVs are therapeutic in a sheep model of sepsis following smoke inhalation and Pseudomonas aeruginosa pneumonia. We hypothesize that MSC MVs will improve oxygenation (PaO2/FiO2) by restoring alveolar fluid clearance, reducing lung protein permeability and alveolar inflammation, increasing bacterial killing, and decreasing pulmonary vascular pressure by releasing nitric oxide and restoring the glycocalyx layer on the endothelium. The overall goal of the current submission is to further develop the biological rationale for using MSC MVs in acute respiratory distress syndrome in anticipation for a possible FDA IND submission.

项目摘要/摘要 急性呼吸窘迫综合征仍然是一种破坏性综合征，影响了20万多名患者 在美国，死亡率接近40％。目前，没有药物治疗 可降低死亡率。因此，需要创新的疗法。在各种临床前模型中 急性肺损伤，间充质干细胞（MSC）已被证明是秘密多种旁分泌因子 可以降低肺内皮和上皮渗透性，降低注射，增强组织修复和 抑制细菌生长，最终降低死亡率。尽管在早期临床试验中具有良好的安全性，但 但是，MSC具有多次通过后发起的恶性转化的能力 培养以及促进小动物模型中肿瘤生长的能力。在以前的赠款期内，我们 发现由人MSC释放的微泡（MV）与母体在生物学上一样活跃 急性肺损伤的炎症和传染性原因。曾经考虑过细胞碎片，MVS，Anclear 圆形膜结合的蔬菜50 nm至200 nm，这些蔬菜始终由许多细胞类型释放， 现在被认为是细胞 - 细胞通信的重要机制。我们对 当前的建议是MSC MV具有生物活性，并且其在严重细菌中的治疗活性 肺炎是通过转移MRNA，microRNA，蛋白质和/或细胞器的转移来介导的。 MVS到受伤的肺泡。在AIM 1中，我们将通过研究 人类MSC MVS对脂质代谢和ATP结合盒转运蛋白的活性的影响 耐药性相关蛋白（MRP）1，患有严重肺炎的小鼠。我们假设MSC MVS 将抑制单核细胞/巨噬细胞中的MRP1活性，并减少酶促转化和分泌 白细胞（LT）A4至LTC/D/E4，涉及肺蛋白渗透性的白细胞，并增加转化率 进入LTB4，白三苯涉及细菌杀死。在AIM 2中，我们将测试人类MSC MV对网的影响 在一个新型的外体内灌注肺炎损伤的液体转运 炎症损伤受伤的人肺泡上皮II型细胞的原发性培养物。我们假设 MSC MVS将通过恢复肺膜表达来预防肺水肿 主要上皮钠通道，αENAC。在AIM 3中，我们将确定人类MSC MV在A中是否具有治疗性 吸入烟雾后败血症的绵羊模型和铜绿假单胞菌肺炎。我们假设 MSC MVS将通过恢复肺泡液清除率来改善氧合（PAO2/FIO2），从而减少肺蛋白 渗透性和肺泡感染，增加细菌杀伤并减少肺血管 通过释放一氧化氮并在内皮上恢复糖椰子层的压力。总体目标 当前的提交是进一步发展用于在急性呼吸中使用MSC MVS的生物学原理 遇险综合症预期可能提交FDA。