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：
8693007
负责人：
Luis Alberto Ortiz
金额：
$ 38.72万
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/8693007
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，目前尚无成功的治疗方法。实验性矽肺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发挥重要作用。基于干细胞的纤维化肺部疾病治疗尚未实现。我们之前报道过全身施用骨髓间充质干细胞（MSC）可改善实验性肺纤维化。受损肺中保留的间充质干细胞数量很少，这与间充质干细胞通过其组织再生能力促进肺稳态的假设不一致。 MSC 通过影响靶细胞生物学的分泌组表现出重要的旁分泌活性，我们之前报道过 MSC 的有益作用部分是通过其产生限制巨噬细胞炎症的 IL1 受体拮抗剂来介导的。现在我们提供的数据表明，除了可溶性介质外，MSC 还转移线粒体并挤出负载微小 RNA (miRNA) 的外泌体，作为细胞通讯机制，在肺纤维化过程中重新编程先天免疫。我们的中心假设是 MSC 利用微泡作为传递肽、miRNA 和线粒体的手段，以重新编程先天免疫并改善矽肺。我们提出以下具体目标：1) 确定 MSC 衍生的微泡是否解释了 MSC 对二氧化硅刺激的巨噬细胞的影响。 MSC利用微泡的产生来建立与巨噬细胞通讯的细胞独立机制，以重新编程其免疫活性。我们将确定将 MSC 衍生的微泡注射到暴露于二氧化硅的小鼠中是否与完整的 MSC 细胞一样有效地防止 Ly6C/Ghi 巨噬细胞在肺部积聚，从而保护小鼠免受矽肺病的影响。 2) 确定 miRNA 转移在矽肺期间 MSC 介导的先天免疫重编程中的作用。 MSC 衍生的微泡含有 miRNA，并且这些在不同的人类 MSC 中高度保守。我们将确定 MSC 衍生的微泡内的特定 miRNA 转录物是否概括了微泡介导的巨噬细胞重编程，并确定抑制 MSC 中 miRNA 的产生是否消除了巨噬细胞中 MSC 或微泡介导的反应。将进行功能获得和丧失的研究，以确定选定的 miRNA 在调节巨噬细胞功能中的作用。 3) 定义自噬和外壳蛋白复合物 II (COPII) 作为 MSC 线粒体转移决定因素的作用。 MSC 将其线粒体转移到周围的巨噬细胞，并且线粒体样结构存在于 MSC 衍生的微泡中。我们将研究自噬和 COPII 外壳系统是否调节线粒体负载到微泡中。我们将测试以下假设：通过将巨噬细胞与 MSC 衍生的微泡一起孵育，可以重现线粒体转移，并恢复暴露于二氧化硅的巨噬细胞中线粒体复合物 I 的酶活性。最后，我们将确定暴露于二氧化硅的小鼠的肺中是否发生线粒体转移。