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，细胞来源的囊泡可以通过 RNA 种类的转移来改变单独靶细胞的基因组表型。进一步的研究表明，间充质干细胞来源的囊泡（MSC-dv）可以通过 microRNA 的转移介导受损肾脏和骨髓组织的愈合。迫切需要确定 MSC-dv 是否可以用于逆转患有此类疾病的人类的肾脏或骨髓损伤。目前的建议是确定分离和表征MSC-dv的最佳方法，该MSC-dv携带“愈合”microRNA到因甘油暴露或缺血/再灌注而损伤的肾组织或因辐射而损伤的骨髓组织（干细胞）。然后，将通过深度测序确定“治愈”MSC-dv 中的 microRNA，并根据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来确定特定的“治愈”microRNA。这将通过脂质转染受损细胞系并分析增殖和凋亡来进行。我们还将描述用特定的“治疗”microRNA 加载“治疗”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 micro RNA 在组织修复中的作用，2.) 负责细胞命运变化的特定 micro RNA 或 microRNA 子集的定义，以及 3.) 开发一种递送 MSC 囊泡 microRNA 以恢复受损肾脏或骨髓组织的方法。这构成了该提案的第一阶段。在第 2 阶段，在确定 MCS-dv 中最佳包装的“愈合”microRNA 用于递送至受损骨髓或肾组织后，我们接下来需要优化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“愈合”表型的稳定性。最佳愈合水平将通过时间过程实验进行评估，其中评估不同时间和频率的输注在我们的模型中的愈合效果。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，我们将在最佳输注计划后跟踪组织分布，并确定 MSC-dv 在不同条件下随时间的稳定性。第一阶段和第二阶段概述的研究总体来说是独特且创新的。 MSC-dv 中递送 microRNA 来治愈组织损伤旨在为以前未尝试过的不同肾脏和骨髓疾病的治疗干预奠定基础。