Proangiogenic microRNA in Microvesicles Released from Adipose-derived Stem Cells

脂肪干细胞释放的微泡中的促血管生成 microRNA

• 批准号: 8642653
• 负责人: Dong Liu
• 金额: $ 14.15万
• 依托单位: MOREHOUSE SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-04-10 至 2015-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8642653
• 关键词: 3' Untranslated Regions; Address; Adipose tissue; Animal Model; Animals; Attention; Binding; Biological Assay; Cardiac; Cell physiology; Cells; Cellular biology; Cerebrum; Clinical; Clinical Trials; Complement; Disease; Distant; Endocrine; Endothelial Cells; Epidemic; Event; Fatty acid glycerol esters; Future; Gene Targeting; Genetic Materials; Goals; Growth Factor; Histocompatibility Antigens; Human; Intervention; Ischemia; Knowledge; Light; Limb structure; Lipids; Location; Luciferases; Mediating; Mediator of activation protein; Medical; Messenger RNA; MicroRNAs; Molecular; Morbidity - disease rate; Myocardial Ischemia; Nucleic Acid Regulatory Sequences; Operative Surgical Procedures; Patients; Play; Process; Property; Protein Microchips; Proteins; Publications; RNA; Recombinant Proteins; Reporter; Reporting; Reverse Transcriptase Polymerase Chain Reaction; Role; Signal Transduction; Small RNA; Source; Stem cells; Stroke; Subfamily lentivirinae; Surface; Therapeutic; Time; Tissues; Transplantation; Tube; Vascular Diseases; Vascular Endothelial Cell; Western Blotting; angiogenesis; base; cell motility; gene therapy; inhibitor/antagonist; innovation; intercellular communication; loss of function; mRNA Expression; migration; mortality; novel; novel therapeutic intervention; paracrine; preconditioning; protein expression; research study; stem cell therapy; transdifferentiation; treatment strategy

DESCRIPTION (provided by applicant): Ischemic cardiac and cerebral vascular diseases continue to represent a significant and growing source of morbidity and mortality despite advances in traditional treatments. Recently, novel emerging therapeutic strategies focus on stem cells. Adipose tissue derived stem cells (ASCs), which are easily acquired and have reduced surface histocompatibility antigens increasing their allo-transplantation potential, were demonstrated to have beneficial effect on ischemic heart and limb in animals likely due to a paracrine function rather than transdifferentiation. Besides secreted soluble growth factors, cell-released microvesicles (MVs) have been recently described as a new mechanism of intercellular communication. MVs play an important role in cell biologic processes not only by specifically targeting recipient cells to deliver proteins, lipids and/or trigger downstream signalng events, but also by transferring genetic material, mRNA and microRNA. Our preliminary studies for the first time demonstrated that 1). MVs are released from human ASCs (hASCs) and promote vascular endothelial cell migration, 2). They contain RNA, mostly small RNA, 3). These small RNAs, rich in angiogenesis-regulating miRNAs, are proangiogenic. Although the importance of cognate miRNA in angiogenesis and endothelial function has been addressed, the mechanism for of angiogenic effect of miRNAs present in stem cell-released MVs is so far unknown. The goal of this proposal is to unravel the mechanistic aspects of the proangiogenic effect of hASCs-MVs in detail. We hypothesize that microRNAs in MVs secreted by hASCs is proangiogenic. In Aim 1 we will examine the angiogenic effects of miRNAs in hASCs-released MVs using two loss-of-function strategies. hASCs will be transduced with Lentivirus-based specific anti-miRNA hairpin expression construct and the angiogenic potential of the ensuing MVs on the endothelial cells will be assessed. Vascular endothelial cells will be co-transfected with specific anti-miRNA inhibitor and small RNAs isolated from hASCs-released MVs and assessed for angiogenesis. In Aim 2, we will determine the target genes of proangiogenic miRNA from hASCs-released MVs in vascular endothelial cells by profiling analysis of mRNA and proteins. The predicted target genes of miRNA from hASCs-released MVs in recipient cells will be confirmed by using a 3'UTR insertion reporter constructs. These studies will expand our currently scant knowledge of mechanistic aspects of proangiogenic miRNA present in stem cell-released MVs as well as shed light on their paracrine/endocrine properties and set the basis for their use as a novel therapeutic approach for cerebro-vascular disease.

描述（由申请人提供）：尽管传统治疗方面取得了进步，但缺血性心脏和脑血管疾病仍然代表着发病率和死亡的重要来源。最近，新的新兴治疗策略集中在干细胞上。脂肪组织衍生的干细胞（ASC）很容易获得，并且降低了表面组织相容性抗原增加其同种转移潜力，被证明对动物的缺血性心脏和肢体具有有益的作用，可能是由于旁分泌功能而不是转化差异。除了分泌的可溶性生长因子外，细胞释放的微泡（MV）最近被描述为细胞间通信的新机制。 MVS在细胞生物过程中起着重要作用，不仅是通过专门针对受体细胞来传递蛋白质，脂质和/或触发下游信号事件，还通过转移遗传材料，mRNA和microRNA来发挥了重要作用。我们的初步研究首次证明了1）。 MV从人类ASC（HASC）释放，并促进血管内皮细胞迁移，2）。它们包含RNA，主要是小RNA，3）。这些富含血管生成调节的miRNA的小RNA具有促血管生成。尽管已经解决了同源miRNA在血管生成和内皮功能中的重要性，但迄今为止，尚不清楚干细胞释放的MVS中存在的miRNA的血管生成作用机制。该提案的目的是详细阐明HASCS-MV的促血管生成作用的机械方面。我们假设HASC分泌的MVS中的microRNA具有促生态性。在AIM 1中，我们将使用两种功能丧失策略检查miRNA在HASCS释放的MV中的血管生成作用。将通过基于慢病毒的特异性抗MIRNA发夹表达构建体转导HASC，并评估随后的MV在内皮细胞上的血管生成潜力。血管内皮细胞将与特异性抗MIRNA抑制剂共转染，并从HASCS释放的MVS分离并评估血管生成。在AIM 2中，我们将通过分析mRNA和蛋白质分析血管内皮细胞中HASCS释放的MVS的促血管生成miRNA的靶基因。通过使用3'UTR插入报告基因构建体，将确认来自HASCS释放的MVS中miRNA的预测靶基因。这些研究将扩大我们目前对存在于干细胞释放的MV中的促血管生成miRNA的机械方面的知识，并阐明了其旁分泌/内分泌特性，并为它们用作大脑血管疾病的新型治疗方法树立了基础。