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: 8581373
• 负责人: PETER J. QUESENBERRY
• 金额: $ 25.66万
• 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 2015-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8581373
• 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; Functional RNA; Genomics; Glycerol; Healed; 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; Vesicle; Work; base; cohort; deep sequencing; density; design; extracellular; healing; injured; innovation; intravenous drip; irradiation; kidney cell; preclinical study; public health relevance; 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转移RNA物种改变单独靶细胞的基因组表型。进一步的工作表明，间充质干细胞衍生的囊泡（MSC-DV）可以通过转移microRNA介导受伤的肾脏和骨髓组织的愈合。一个关键的需求是确定MSC-DV是否可以用于扭转如此苦难的人类的肾脏或骨髓损害。目前的建议是定义最佳方法，用于分离和表征MSC-DV携带“愈合” microRNA的MSC-DV与甘油暴露或通过辐照受伤受伤的缺血/再灌注或骨髓组织（干细胞）伤害的肾脏组织。然后，“愈合” MSC-DV中的microRNA将通过深层测序和特定的“愈合” microRNA确定，取决于其将其交付到受伤的肾脏和骨髓细胞系模型中并恢复增殖并减少凋亡的能力确定。这将通过Lipofect损伤细胞系并分析增殖和凋亡来进行。我们还将表征使用特定“愈合” microRNA加载“治愈” MSC-DV的最佳方法。预计这些研究将产生以下预期结果：1。）了解微RNA在组织恢复中的作用的理解，2。）。2。）对细胞命运变化的特定微RNA或microRNA的子集的定义，以及3.）开发MSC MicroRNA的方法，用于恢复受伤的肾脏肾脏或骨髓组织。这构成了提案的第1阶段。在第2阶段，在确定MCS-DV中最佳包装的“愈合” microRNA以递送到受伤的骨髓或肾脏组织之后，我们接下来需要优化组织修复的输送时间表，评估早期和晚期毒性，并分布INFOUDER剂。此外，我们需要评估具有储存的MicroRNA富MSC-D囊泡“愈合”表型的稳定性。最佳的愈合水平将在时间过程中评估，其中评估了不同时间和输注频率以在我们的模型中评估愈合效应。在这些研究过程中，我们还将确定注入动物的早期和晚期毒性。总动物器官组织学和生存率也将被评估。 接下来，我们将在最佳输注时间表后跟踪组织分布，并在不同条件下确定MSC-DV的稳定性。第1阶段和第2阶段概述的整体研究都是独特而创新的。 MSC-DV中microRNA的递送以治愈组织损伤，旨在为以前尚未尝试过的不同肾脏和骨髓疾病的治疗干预措施做好准备。