FAS Controls Exosome-Mediated miRNA Transfer in MSC-Based Therapy

FAS 在基于 MSC 的治疗中控制外泌体介导的 miRNA 转移

• 批准号: 9243614
• 负责人: Chi-Der Chen
• 金额: $ 7.08万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-07 至 2018-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9243614
• 关键词: Adenoviruses; Autoimmune Diseases; Autoimmune Responses; Award; Biological Assay; Blood Vessels; Bone Marrow; Bone Resorption; Cell Lineage; Cell Therapy; Cells; Cellular Structures; Chronic; Clinical; Collagen; Communication; Complex; Connective Tissue Diseases; Data; Deposition; Development; Disease; Disease Management; Exocytosis; FRAP1 gene; Fracture; Functional disorder; Goals; Head and neck structure; High Prevalence; Homeostasis; Human; Immune; Immunoprecipitation; In Vitro; Infiltration; Infusion procedures; Interleukin 4 Receptor; Knockout Mice; Knowledge; Label; Mandible; Mediating; Membrane Protein Traffic; Mentorship; Mesenchymal Stem Cell Transplantation; Mesenchymal Stem Cells; MicroRNAs; Mus; Organ; Osteogenesis; Osteopenia; Osteoporosis; Osteoporotic; Outcome; Pathogenesis; Pathogenicity; Patients; Phase; Phenotype; Play; Production; Property; Regulation; Research; Research Personnel; Role; Scleroderma; Sclerosis; Signal Transduction; Skin; Small Interfering RNA; Stem cells; Supervision; Systemic Scleroderma; Th2 Cells; Therapeutic; Therapeutic Effect; Tissues; Training; Transplantation; United States National Institutes of Health; Vascular Diseases; Vertebral Bone; Vesicle; bone loss; career; chemokine; clinical application; collaborative environment; craniofacial; cytokine; disease phenotype; epigenetic regulation; exosome; gene therapy; human disease; immunoregulation; improved; in vivo; inhibitor/antagonist; knock-down; lipid biosynthesis; microscopic imaging; migration; neutrophil; novel; novel therapeutics; overexpression; public health relevance; skills; skin disorder; stem cell differentiation; stem cell therapy; trafficking; translational study; tsk mouse

Project Description Systemic sclerosis/scleroderma (SSc) is a connective tissue disease characterized by vascular, immune and fibrotic changes in the skin and internal organs. SSc patients are known to have high prevalence of osteopenia, but the pathophysiological mechanism causing this osteoporosis in SSc patients is unknown. Systemic infusion of mesenchymal stem cells (MSCs) yields a therapeutic effect in a variety of autoimmune diseases; however, the reasons behind this MSC-based therapy are not fully understood. As a secretome, MSCs release large amounts of cytokines and small vesicles for immunoregulation and communication with surrounding cells. The goal of this proposal is to investigate how small vesicles are controlled by a set of membrane traffic proteins for cell component reuse in MSC transplantation (MSCT). My preliminary data show that MSCT significantly rescues disease phenotypes in SSc mice through rebuilding tissue homeostasis and lineage commitment of host MSCs. miRNA-151-5p is reused by recipient MSCs through exosome trafficking resulting in markedly recovered lineage selection of host MSCs. A micro-vesicle trafficking complex assembled in donor MSCs regulated exocytosis for exosome-mediated miRNA transfer. In addition, developmental endothelial locus-1 (Del-1), an immune suppressive molecule, expresses in MSCs and releases via exosomes to inhibit neutrophil migration. The central hypothesis of this proposal is that cell component transferred from donor to recipient MSCs ameliorates autoimmune disease phenotypes through FAS-assembled exocytotic machinery. During the K99 phase, I will explore the efficacy of MSCT in SSc mice and uncover the interplay between immune cells and tissue-specific stem cells to elucidate the pathophysiological mechanism of SSc (Aim 1; K99). Since miRNA-151-5p has been identified to transfer from donor to recipient MSCs, I will determine the function of miRNA-151-5p in MSC differentiation and use miRNA-151-5p gene therapy in SSc mice (Aim 2; K99). During the R00 phase, I will determine the role of the exosome in MSCT by direct infusion of exosomes for disease treatment and examine the role of the exocytosis molecule Rab27a in regulation of exosome release. I will identify a novel micro-vesicle trafficking complex from MSCs and determine its function in controlling cell component reuse (Aim 3; R00). Finally, I will examine the therapeutic effect of Del-1, which can be released via exosomes from donor MSCs to restrain cytokine production and disease phenotypes (Aim 4; R00). Upon successful completion of the Specific Aims, this translational study will extend our knowledge of SSc pathogenesis and describe detailed mechanisms of MSC-based therapy. Under my primary mentorship of Drs. Hajishengallis and Shi, the training plan will provide the opportunity to extend and develop the necessary knowledge and skills within a collaborative environment. An identified outstanding mentorship team and collaborators will broaden my training in different research fields. Under their supervision and guidance, this NIH K99/R00 Award will largely improve my ability to begin my scientific career as an independent investigator.

项目描述 全身性硬化/硬皮病（SSC）是一种结缔组织疾病，其特征是血管，免疫和 皮肤和内部器官的纤维化变化。已知SSC患者的骨质减少症患病率很高， 但是，导致SSC患者这种骨质疏松症的病理生理机制尚不清楚。全身输注 间充质干细胞（MSC）在多种自身免疫性疾病中产生治疗作用；然而， 这种基于MSC的治疗的原因尚未完全了解。作为一个秘密，MSC释放大型 用于免疫调节和与周围细胞通信的细胞因子和小囊泡量。这 该建议的目标是研究如何通过一组膜交通蛋白来控制小囊泡 MSC移植（MSCT）中的细胞组件再利用。我的初步数据表明MSCT显着 通过重建组织稳态和谱系承诺，从而在SSC小鼠中营救疾病表型 主持人MSC。 MiRNA-151-5p通过外部贩运重复使用MSC，导致明显 恢复主机MSC的谱系选择。在供体MSC中组装的微维型贩运复合物 调节外胞吐作用，用于外泌体介导的miRNA转移。此外，发育性内皮基因座-1 （DEL-1）是一种免疫抑制分子，在MSC中表达并通过外泌体释放以抑制中性粒细胞 迁移。该提议的中心假设是细胞成分从供体转移到 接受者MSC通过FAS组装的外胞毒性改善自身免疫性疾病表型 机械。在K99阶段，我将探索MSCT在SSC小鼠中的功效并发现相互作用 免疫细胞和组织特异性干细胞之间以阐明SSC的病理生理机理 （AIM 1; K99）。由于已经确定了MiRNA-151-5P从捐助者转移到受体MSC，我将 确定miRNA-151-5p在MSC分化中的功能，并在SSC中使用miRNA-151-5p基因治疗 小鼠（AIM 2; K99）。在R00阶段，我将通过直接输注来确定外泌体在MSCT中的作用 外泌体进行疾病治疗，并检查胞吐分子Rab27a在调节中的作用 外泌体释放。我将从MSC中确定一种新型的微维化合物，并确定其功能 在控制单元组件的重复使用时（AIM 3; R00）。最后，我将研究Del-1的治疗作用， 可以通过供体MSC的外泌体释放以限制细胞因子的产生和疾病表型（AIM 4; R00）。成功完成具体目标后，这项翻译研究将扩展我们对 SSC发病机理并描述基于MSC的治疗的详细机制。在我的主要指导下 博士。 Hajishengallis和Shi，培训计划将为扩展和开发必要的机会提供机会 在协作环境中的知识和技能。确定的杰出指导团队和 合作者将扩大我在不同研究领域的培训。在他们的监督和指导下， NIH K99/R00奖将在很大程度上提高我开始从事独立研究者科学生涯的能力。