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

基本信息

批准号：
9906197
负责人：
Chi-Der Chen
金额：
$ 24.41万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-04-03 至 2022-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9906197
关键词：
Adenoviruses Autoimmune Diseases Autoimmune Responses Award Biological Assay Blood Vessels Bone Marrow Bone Resorption Cell Differentiation process Cell Lineage Cell Therapy Cells Cellular Structures Chronic Clinical Collagen Communication Complex Connective Tissue Diseases Data Deposition Development Disease Disease Management Endothelium Exocytosis FRAP1 gene Fracture Functional disorder Goals Head and neck structure High Prevalence Homeostasis Human Immune Immunoprecipitation Impairment In Vitro Infiltration Infusion procedures Interleukin 4 Receptor Knockout Mice Knowledge Label Mandible Mediating Membrane 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 Supervision Systemic Scleroderma Th2 Cells Therapeutic Therapeutic Effect Tissues Training United States National Institutes of Health Vascular Diseases Vertebral Bone Vesicle bone loss career chemokine clinical application collaborative environment conditional knockout 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 stem cells trafficking translational study tsk mouse

项目摘要

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.

项目描述系统性硬化症/硬皮病（SSc）是一种结缔组织疾病，其特征是血管、免疫和皮肤和内脏器官的纤维化变化。众所周知，SSc 患者骨质减少的患病率很高，但引起 SSc 患者骨质疏松的病理生理机制尚不清楚。全身输注间充质干细胞（MSC）对多种自身免疫性疾病产生治疗作用；然而，这种基于 MSC 的疗法背后的原因尚不完全清楚。作为分泌组，间充质干细胞释放大量大量的细胞因子和小囊泡用于免疫调节和与周围细胞的通讯。这该提案的目标是研究小囊泡如何由一组膜运输蛋白控制间充质干细胞移植（MSCT）中的细胞成分重复使用。我的初步数据显示 MSCT 显着通过重建组织稳态和谱系承诺来拯救 SSc 小鼠的疾病表型宿主间充质干细胞。 miRNA-151-5p 通过外泌体运输被受体 MSC 重复利用，从而显着恢复宿主 MSC 的谱系选择。在供体间充质干细胞中组装的微囊泡运输复合体调节外泌体介导的 miRNA 转移的胞吐作用。此外，发育内皮基因座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。该提案的中心假设是细胞成分从供体转移到受体 MSC 通过 FAS 组装的胞吐作用改善自身免疫性疾病表型机械。在 K99 阶段，我将探索 MSCT 对 SSc 小鼠的功效并揭示其相互作用免疫细胞和组织特异性干细胞之间的关系，以阐明 SSc 的病理生理机制（目标 1；K99）。由于 miRNA-151-5p 已被确定可以从供体 MSC 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（目标 2；K99）。在R00阶段，我将通过直接输注来确定外泌体在MSCT中的作用外泌体用于疾病治疗并检查胞吐分子 Rab27a 在调节中的作用外泌体释放。我将从间充质干细胞中鉴定出一种新型微泡运输复合物并确定其功能控制单元组件重用（目标 3；R00）。最后，我将检查Del-1的治疗效果，它可以通过供体 MSC 的外泌体释放，以抑制细胞因子的产生和疾病表型（目的 4； R00）。成功完成具体目标后，这项转化研究将扩展我们的知识 SSc 发病机制并描述基于 MSC 的治疗的详细机制。在我的主要指导下博士。 Hajishengallis 和 Shi 表示，培训计划将提供扩展和发展必要的机会协作环境中的知识和技能。一支公认的优秀导师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 奖将极大地提高我作为独立研究者开始科学生涯的能力。