Regulation of renal and bone marrow injury by extracellular vesicle non-coding RN

细胞外囊泡非编码RN对肾和骨髓损伤的调节

• 批准号: 9128771
• 负责人: PETER J. QUESENBERRY
• 金额: $ 31.87万
• 依托单位: RHODE ISLAND HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-01 至 2018-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9128771
• 关键词: Animal Organ; Animals; Apoptosis; Bone Marrow; Cardiac; Cell Culture System; Cell Line; Cell physiology; Cells; Conditioned Culture Media; Development; Disease; Dose; Drug or chemical Tissue Distribution; Engineering; Epitopes; Frequencies; Genomics; Glycerol; Healed; Health; Histology; Human; In Vitro; Individual; Infusion procedures; Injury; Ischemia; Kidney; Lead; Liver; Lung; Marrow; Mediating; Mesenchymal Stem Cells; Messenger RNA; MicroRNAs; Modeling; Monitor; Mus; Normalcy; Outcome; Phenotype; Ploidies; Population; Pre-Clinical Model; Preparation; Proteins; Proteomics; RNA; Regulation; Renal Tissue; Reperfusion Therapy; Role; Schedule; Staging; Stem cells; Surface; Tail; Testing; Therapeutic Intervention; Time; Tissues; Toxic effect; Transfer RNA; Untranslated RNA; Vesicle; Work; base; cohort; deep sequencing; density; design; exosome; extracellular vesicles; healing; injured; innovation; intravenous drip; irradiation; kidney cell; preclinical study; relating to nervous system; repaired; research study; response; restoration; time interval; tissue repair

DESCRIPTION (provided by applicant): Recent studies indicate that cellular derived vesicles can alter the genomic phenotype of separate target cells by transfer of RNA species. Further work has indicated that mesenchymal stem cell-derived vesicles (MSC-dv) can mediate healing of injured renal and marrow tissue by transfer of microRNA. A critical need is to determine whether MSC-dv can be utilized to reverse kidney or marrow damage in humans so afflicted. The present proposal is to define the optimum approach for separating and characterizing MSC-dv carrying "healing" microRNA to renal tissue injured by glycerol exposure or by ischemia/reperfusion or marrow tissue (stem cells) injured by irradiation. The microRNA in "healing" MSC-dv will then be determined by deep sequencing and the specific "healing" microRNA determined by its capacity to be delivered to injured renal and marrow cell line models and restore proliferation and reduce apoptosis. This will be carried out by lipofecting injured cell lines and analyzing proliferation and apoptosis. We will also characterize optimal approaches to loading "healing" MSC-dv with specific "healing" microRNA(s). It is anticipated that these studies will yield the following expected outcomes: 1.) An understanding of the role of micro RNAs in tissue restoration, 2.) a definition of the specific micro RNA or subset of microRNAs responsible for cell fate change and 3.) the development an approach for the delivery of MSC vesicle microRNA to restore injured renal or marrow tissue. This constitutes Stage 1 of the proposal. In Stage 2, after the determination of a "healing" microRNA packaged optimally in MCS-dv for delivery to injured marrow or renal tissue, we next need to optimize delivery schedule for tissue repair, evaluate early and late toxicity, and distribution of the infued agent. In addition, we need to evaluate the stability of "healing" phenotype of the microRNA rich MSC-d vesicles with storage. Optimal healing levels will be assessed in time course experiments where different times and frequencies of infusion are evaluated as to healing effects in our models. During the course of these studies we will also determine early and late toxicities in the infused animals. Total animal organ histology and survival will also be assessed. Next we will track tissue distribution after an optimal infusion schedule and determine the stability of MSC-dv over time under different conditions. The totality of the studies outlined in Stage 1 and 2 are unique and innovative. The delivery of microRNA in MSC-dv to heal tissue injuries is designed to prepare the basis for therapeutic interventions in different renal and marrow diseases which have not been previously attempted.

描述（由申请人提供）：最近的研究表明，细胞衍生的囊泡可以通过RNA种类的转移改变单独靶细胞的基因组表型。进一步的研究表明，间充质干细胞衍生的囊泡（MSC-DV）可以通过转移microRNA介导损伤的肾脏和骨髓组织的愈合。一个关键的需要是确定MSC-dv是否可以用于逆转受此折磨的人类的肾脏或骨髓损伤。目前的建议是定义分离和表征MSC-dv的最佳方法携带“愈合”microRNA的甘油暴露或缺血/再灌注损伤的肾组织或骨髓组织（干细胞）辐射损伤。然后通过深度测序确定“愈合”MSC-dv中的microRNA，并通过其递送至受损肾和骨髓细胞系模型并恢复增殖和减少凋亡的能力确定特异性“愈合”microRNA。这将通过脂质转染损伤的细胞系并分析增殖和凋亡来进行。我们还将表征用特异性“愈合”microRNA装载“愈合”MSC-dv的最佳方法。预计这些研究将产生以下预期结果：1.理解微小RNA在组织修复中的作用，2。负责细胞命运改变的特定微小RNA或微小RNA子集的定义，以及3.）开发一种用于递送MSC囊泡microRNA以恢复受损的肾或骨髓组织的方法。这是提案的第一阶段。在第2阶段，在确定了最佳包装在MCS-dv中以递送至受损骨髓或肾组织的“愈合”microRNA之后，我们接下来需要优化用于组织修复的递送时间表，评估早期和晚期毒性以及输注剂的分布。此外，我们需要评估富含microRNA的MSC-d囊泡在储存时的“愈合”表型的稳定性。最佳愈合水平将在时程实验中评估，其中不同的输注时间和频率对我们模型中的愈合效果进行了评估。在这些研究过程中，我们还将确定输注动物的早期和晚期毒性。还将评估总动物器官组织学和存活率。 接下来，我们将在最佳输注时间表后跟踪组织分布，并确定不同条件下MSC-dv随时间的稳定性。第一阶段和第二阶段概述的所有研究都是独特和创新的。在MSC-dv中递送microRNA以愈合组织损伤旨在为先前未尝试过的不同肾脏和骨髓疾病的治疗干预奠定基础。