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.

项目概要/摘要 急性呼吸窘迫综合征仍然是一种破坏性综合征，影响超过 200,000 名患者 在美国，每年的死亡率接近40%。目前尚无药物治疗 可以降低死亡率。因此，需要创新疗法。在各种临床前模型中 急性肺损伤时，间充质干细胞（MSC）已被证明可以分泌多种旁分泌因子， 可以降低肺内皮和上皮的通透性，减少炎症，增强组织修复， 抑制细菌生长，最终降低死亡率。尽管早期临床试验的安全性良好， 然而，间充质干细胞在多次传代后具有自发恶性转化的能力 培养以及促进小动物模型中肿瘤生长的能力。在之前的资助期内，我们 发现人类 MSC 释放的微泡 (MV) 与亲本干细胞具有相同的生物活性 急性肺损伤的炎症和感染原因。曾经被认为是细胞碎片、MV、无核细胞 大小为 50 nm 至 200 nm 的圆形膜结合囊泡，由许多细胞类型组成型释放， 现在被认为是细胞间通讯的重要机制。我们的总体假设 目前的建议是，MSC MV 具有生物活性，其对严重细菌的治疗活性 肺炎是通过 mRNA、microRNA、蛋白质和/或细胞器的转移介导的 MVs 到受伤的肺泡。在目标 1 中，我们将通过研究 人 MSC MV 对脂质代谢和 ATP 结合盒转运蛋白活性的影响，多药 抵抗相关蛋白（MRP）1，在患有严重肺炎的小鼠中。我们假设 MSC MV 会抑制单核细胞/巨噬细胞中的 MRP1 活性，并减少酶的转化和分泌 白三烯 (LT)A4 转为 LTC/D/E4，白三烯参与肺蛋白通透性，并增加转化 转化为LTB4，白三烯参与细菌杀灭。在目标2中，我们将测试人类MSC MV在网络上的效果 患有严重细菌性肺炎的新型离体灌注人肺中的液体运输以及 因炎症损伤而损伤的人肺泡上皮 II 型细胞的原代培养物。我们假设 MSC MV 将通过恢复根尖膜表达来预防肺水肿形成 主要上皮钠通道，αENaC。在目标 3 中，我们将确定人类 MSC MV 是否具有治疗作用 吸入烟雾后败血症和铜绿假单胞菌肺炎的绵羊模型。我们假设 MSC MV 将通过恢复肺泡液清除率、减少肺蛋白来改善氧合 (PaO2/FiO2) 通透性和肺泡炎症，增加细菌杀灭，减少肺血管 通过释放一氧化氮并恢复内皮上的糖萼层来消除压力。总体目标为 当前提交的内容是进一步开发在急性呼吸道疾病中使用 MSC MV 的生物学原理 预期可能向 FDA 提交 IND 的痛苦综合症。