Mesenchymal stem cell secretome in lung fibrosis: mitochondria and RNA shuttle

肺纤维化中的间充质干细胞分泌组：线粒体和RNA穿梭

基本信息

批准号：
8446569
负责人：
Luis Alberto Ortiz
金额：
$ 38.58万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-07-01 至 2017-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8446569
关键词：
Accounting Adoptive Transfer Aging Alzheimer&apos s Disease Anti-Inflammatory Agents Anti-inflammatory Autophagocytosis Biology Bone Marrow Breathing Capsid Proteins Cardiovascular Diseases Cell Communication Cell Therapy Cells Centers for Disease Control and Prevention (U.S.)Communication Complex Data Disease Endocytic Vesicle Eukaryotic Cell Fibrosis Figs - dietary Gene Expression Hamman-Rich syndrome Homeostasis Human ITGAM gene Immune In Vitro Inflammation Inflammatory Interleukin-1 Receptors Interleukin-10 Link Lung Lung diseases Malignant Neoplasms Mediating Mediator of activation protein Mesenchymal Stem Cells MicroRNAs Mitochondria Mitochondrial RNA Modeling Mus Natural Immunity Organelles Parkinson Disease Pathogenesis Pennsylvania Peptides Play Population Production Publishing Pulmonary Fibrosis Regulation Reporting Role Silicon Dioxide Silicosis Stem cells Structure System Testing Transcript Vesicle gene environment interaction human DICER1 protein in vivo injured loss of function lung injury macrophage mitochondrial dysfunction monocyte origen paracrine peripheral blood prevent protein complex public health relevance response tissue regeneration

项目摘要

DESCRIPTION (provided by applicant): Pulmonary fibrosis is a lethal disease for which no successful therapy is available. Experimental silicosis is an excellent model to study this disease. In the US, more than 2 million people are exposed to silica every year. Macrophages play a fundamental role in lung fibrosis. Stem cell-based therapy for fibrotic lung diseases has yet to be realized. We previously reported that systemic administration of bone marrow derived mesenchymal stem cells (MSCs) ameliorates experimental lung fibrosis. The number of MSCs retained in the injured lung is small and inconsistent with the postulate that MSCs promote lung homeostasis by their capacity for tissue regeneration. MSCs demonstrate an important paracrine activity through a secretome that influences the biology of target cells and we previously reported that beneficial effects of MSCs are mediated, in part, by their production of IL1 receptor antagonist that limits macrophage inflammation. Now we present data that in addition to soluble mediators, MSC transfer mitochondria and extrude micro RNA (miRNA) ladened exosomes as a mechanism of cell communication to reprogram the innate immunity during lung fibrosis. Our central Hypothesis is that MSCs employ microvesicles as a mean to deliver peptides, miRNAs, and mitochondria to reprogram the innate immunity and ameliorate silicosis. We propose the following specific Aims: 1) To determine whether MSC-derived microvesicles account for the effects of MSC on silica stimulated macrophages. MSC use micro vesicle production to establish a cell independent mechanism of communication with macrophages to reprogram their immune activity. We will determine whether administration of MSC-derived microvesicles into silica-exposed mice is as effective as intact MSC cells in preventing accumulation of Ly6C/Ghi macrophages in the lung thus protecting mice from silicosis. 2) To determine the role of miRNA transfer in MSC-mediated reprogramming of innate immunity during silicosis. MSC-derived microvesicles contain miRNAs and these are highly conserved among different human MSCs. We will determine whether specific miRNA transcripts inside MSC-derived microvesicles recapitulate the micro vesicle-mediated reprogramming of macrophages and determine if inhibiting miRNA production in MSCs eliminates MSC or micro vesicle-mediated responses in macrophages. Gain and loss-of-function studies will be done to define the role of select miRNAs in regulating macrophage function. 3) To define the role of autophagy and coat protein complex II (COPII) as determinants of MSC mitochondrial transfer. MSCs transfer their mitochondria to surrounding macrophages and that mitochondria like structures are present in MSC-derived microvesicles. We will examine whether autophagy and COPII coat systems regulate the load of mitochondria into microvesicles. We will test the hypothesis that mitochondrial transfer could be recapitulated by incubation of macrophages with MSC-derived microvesicles and restore the enzymatic activity of mitochondrial complex I in silica-exposed macrophages. Finally, we will determine whether transfer of mitochondria occurs in the lung of silica-exposed mice.

描述（由申请人提供）：肺纤维化是一种致命疾病，没有成功的治疗。实验硅化是研究这种疾病的绝佳模型。在美国，每年有超过200万人接触二氧化硅。巨噬细胞在肺纤维化中起着基本作用。基于干细胞的纤维化肺疾病疗法尚未实现。我们先前报道说，骨髓的全身给药衍生的间充质干细胞（MSC）可以改善实验性肺纤维化。保留在受伤的肺中的MSC数量很小，并且与MSC通过组织再生的能力促进肺稳态的假设不一致。 MSC通过一种影响靶细胞的生物学的分泌组表现出重要的旁分泌活性，我们先前报道说，MSC的有益作用是通过其产生IL1受体拮抗剂的一部分介导的，从而限制了巨噬细胞炎症。现在，我们提出的数据表明，除了可溶性介质外，MSC转移线粒体和挤压微RNA（miRNA）放大的外泌体是细胞通信的一种机制，可以重新编程肺纤维化过程中先天免疫。我们的中心假设是，MSC采用微囊泡作为输送肽，miRNA和线粒体的平均值来重新编程先天免疫和改善硅化作用。我们提出以下特定目的：1）确定MSC衍生的微泡是否解释了MSC对二氧化硅刺激巨噬细胞的影响。 MSC使用微囊泡的产生来建立与巨噬细胞通信的独立细胞机制，以重新编程其免疫活性。我们将确定在暴露于二氧化硅暴露的小鼠中的MSC衍生的微泡是否与完整的MSC细胞一样有效，可以防止LY6C/GHI巨噬细胞在肺部积累，从而保护小鼠免受硅氧化症的影响。 2）确定miRNA转移在MSC介导的硅化过程中先天免疫的重编程中的作用。 MSC衍生的微泡包含miRNA，它们在不同的人类MSC中是高度保守的。我们将确定MSC衍生的微泡中的特定miRNA转录本是否概括了巨噬细胞的微囊泡介导的重编程，并确定在MSC中抑制miRNA的产生是否会消除巨噬细胞中MSC或微囊泡介导的反应。将进行功能丧失研究，以定义某些miRNA在调节巨噬细胞功能中的作用。 3）定义自噬和外套蛋白质复合物II（COPII）作为MSC线粒体转移的决定因素。 MSC将其线粒体转移到周围的巨噬细胞中，并且在MSC衍生的微泡中存在类似的线粒体结构。我们将检查自噬和COPII外套系统是否调节线粒体的负载到微泡中。我们将测试以下假设：通过与MSC衍生的微泡孵育巨噬细胞，可以概括线粒体转移，并在暴露于二氧化硅暴露的巨噬细胞中恢复线粒体复合物I的酶活性。最后，我们将确定线粒体的转移是否发生在二氧化硅暴露的小鼠的肺中。