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: 8830995
• 负责人: Jae Woo Lee
• 金额: $ 38.05万
• 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 2016-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8830995
• 关键词: Actins; Acute Lung Injury; Acute Renal Failure with Renal Papillary Necrosis; Acute respiratory failure; Affect; Alveolar; Alveolar Macrophages; Animals; Anti-Inflammatory Agents; Anti-inflammatory; Apical; Beds; Biological; Biology; Bone Marrow; CD44 gene; Cell Therapy; Cell membrane; Cells; Clinical Trials; Critical Illness; Data; Development; Edema; Endothelial Cells; Endothelium; Endotoxins; Epithelial; Epithelial Cells; Epithelium; Escherichia coli; Fibroblasts; Fluid Balance; Growth; Growth Factor; Guanosine Triphosphate Phosphohydrolases; Human; Human Activities; In Vitro; Infection; Inflammation; Inflammatory; Inflammatory Response; Injury; Interferons; Interleukin-1; Kidney; Liquid substance; Lung; Malignant - descriptor; Malignant Neoplasms; Mediating; Membrane; Membrane Proteins; Mesenchymal Stem Cells; Messenger RNA; MicroRNAs; Mitochondria; Modeling; Mus; Organelles; Patients; Permeability; Phenotype; Pre-Clinical Model; Protein Biosynthesis; Proteins; Pulmonary Edema; RNA Synthesis Inhibitors; Receptor Cell; Recovery of Function; Reporting; Research; Research Personnel; Resolution; Role; Safety; Sodium Channel; Stem cells; Stress Fibers; Surface; Syndrome; TNF gene; Testing; Therapeutic; Therapeutic Effect; Therapeutic Uses; Tissues; Translations; Tubular formation; Type II Epithelial Receptor Cell; antimicrobial peptide; apical membrane; cytokine; epithelial Na+ channel; in vivo; in vivo Model; injured; insight; keratinocyte growth factor; lung injury; macrophage; mortality; novel; novel strategies; novel therapeutic intervention; paracrine; pre-clinical; prevent; receptor; repaired; tissue repair; tumor growth

DESCRIPTION (provided by applicant): Acute lung injury (ALI) 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 pharmacologic therapies that reduce mortality. Consequently, further research into translational therapies is needed. Stem cell-based therapy with mesenchymal stem cells (MSC) is one attractive new approach. MSC have the capacity to secrete multiple paracrine factors that can regulate lung endothelial and epithelial permeability, decrease inflammation, enhance tissue repair, and inhibit bacterial growth. In over 150 clinical trials registered with clinicaltrials.gov using MSC as therapy, over 2000 patients have received the cells without any major complications. Despite a favorable safety profile, however, MSC have the capacity for spontaneous malignant transformation following multiple passages in vitro as well as the ability to promote tumor growth in vivo. Recently, some investigators have found that microvesicles (MV) released by human MSC are as biologically active as the stem cells in part through the transfer and expression of MV mRNA in the injured tissue bed. In this application, I propose to study the biology and test the potential therapeutic use of human bone-marrow derived MSC MV as an alternative to cellular therapy in models of ALI. The overall hypothesis is that human MSC MV are biologically active, and that their therapeutic activity is primarily mediated through transfer of mRNA from the MV to injured lung epithelium and lung endothelium. In Aim 1, the primary objective is to study the biology of MSC MV and determine which components of the MSC MV are functionally active, using inhibitors of RNA and protein synthesis and transport. I hypothesize that MSC MV require the transfer of mRNA for key paracrine soluble factors from the MV to the injured lung epithelium or endothelium using a cell membrane receptor, such as CD44, for their therapeutic effect. In Aim 2, I will test the functional activity of human MSC MV on net fluid transport in human alveolar epithelial type II cells and on lung endothelial permeability to protein in human lung microvascular endothelial cells injured by an inflammatory insult, the main pathological features of ALI. I hypothesize that MSC MV will prevent the decrease in net fluid transport in injured type II cells by restoring the apical membrane expression of the major epithelial sodium channel, ¿ENaC, and will reduce the increase in protein permeability in injured lung endothelial cells by preventing the formation of actin stress fibers. In Aim 3, I will determine if human MSC MV are biologically active in mice injured with E.coli endotoxin-induced ALI. I hypothesize that MSC MV will reduce endotoxin-induced ALI in mice by restoring lung endothelial and epithelial protein permeability, lung fluid balance and by reducing alveolar inflammation. These studies will provide novel insights into how MVs are released and the underlying mechanisms that explain why MSC MVs may be effective in tissue repair. Furthermore, the results may provide preclinical data that could facilitate development of MSC MV as a therapy for ALI.

描述（由申请人提供）：急性肺损伤 (ALI) 仍然是一种毁灭性综合征，每年影响美国超过 200,000 名患者，死亡率接近 40%。目前，没有药物疗法可以降低死亡率。因此，需要对转化疗法进行进一步的研究。使用间充质干细胞（MSC）进行干细胞治疗是一种有吸引力的新方法。间充质干细胞具有分泌多种旁分泌因子的能力，这些因子可以调节肺内皮和上皮通透性、减少炎症、增强组织修复和抑制细菌生长。在 ClinicalTrials.gov 注册的 150 多项使用 MSC 作为疗法的临床试验中，超过 2000 名患者接受了细胞治疗，没有出现任何重大并发症。然而，尽管安全性良好，MSC 在体外多次传代后仍具有自发恶性转化的能力，并且具有促进体内肿瘤生长的能力。最近，一些研究人员发现，人间充质干细胞释放的微泡（MV）与干细胞一样具有生物活性，部分是通过MV mRNA在受损组织床中的转移和表达来实现的。在此应用中，我建议研究生物学并测试人骨髓来源的 MSC MV 作为 ALI 模型细胞治疗替代方案的潜在治疗用途。总体假设是人类 MSC MV 具有生物活性，并且其治疗活性主要通过 mRNA 从 MV 转移到受损的肺上皮和肺内皮来介导。目标 1 的主要目标是研究 MSC MV 的生物学，并使用 RNA 和蛋白质合成和运输抑制剂确定 MSC MV 的哪些成分具有功能活性。我假设 MSC MV 需要使用细胞膜受体（例如 CD44）将关键旁分泌可溶性因子的 mRNA 从 MV 转移到受损的肺上皮或内皮，以发挥治疗作用。在目标 2 中，我将测试功能 人MSC MV对人肺泡上皮II型细胞中的净液体运输的活性以及对因炎症损伤而损伤的人肺微血管内皮细胞中的肺内皮对蛋白质的通透性的活性，这是ALI的主要病理特征。我假设 MSC MV 将通过恢复主要上皮钠通道 ENaC 的顶膜表达来防止受损 II 型细胞中净液体转运的减少，并通过阻止肌动蛋白应力纤维的形成来减少受损肺内皮细胞中蛋白质通透性的增加。在目标 3 中，我将确定人 MSC MV 在大肠杆菌内毒素诱导的 ALI 损伤的小鼠中是否具有生物活性。我推测 MSC MV 将通过恢复肺内皮和上皮蛋白通透性、肺液平衡以及减少肺泡炎症来减少内毒素诱导的小鼠 ALI。这些研究将为 MV 如何释放以及解释为什么 MSC MV 在组织修复中有效的潜在机制提供新的见解。此外，结果可能提供临床前数据，促进 MSC MV 作为 ALI 治疗方法的开发。