Therapeutic Potential of Gingival mesenchymal Stem Cell-derived Extracellular Vesicles Enriched with MFG-E8 in Peripheral Nerve Regeneration

富含 MFG-E8 的牙龈间充质干细胞来源的细胞外囊泡在周围神经再生中的治疗潜力

• 批准号: 10042927
• 负责人: Anh D Le
• 金额: $ 20.25万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2022-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10042927
• 关键词: Animals; Anti-Inflammatory Agents; Apoptotic; Applications Grants; Autologous Transplantation; Biological Process; Cell Therapy; Cells; Cerebral Ischemia; Cicatrix; Clinic; Clinical; Clinical Trials; Collagen; Complex; Crush Injury; Custom; Data; Defect; Disease; Epidermal Growth Factor; Evaluation; Factor VIII; Female; Gene Expression; Genes; Genetic; Genetic Transcription; Gingiva; Glycoproteins; Goals; Gold; Histologic; Homeostasis; Human; Hydrogels; Implant; Infection; Inferior; Inflammation; Infusion procedures; JUN gene; Lead; Lipids; Mediating; Mesenchymal Stem Cells; Modeling; Morbidity - disease rate; Mus; Natural regeneration; Nerve; Nerve Regeneration; Neurodegenerative Disorders; Neuroglia; Neuroma; Operative Surgical Procedures; Outcome; Peripheral Nerves; Peripheral nerve injury; Phagocytosis; Pharmacology; Phenotype; Play; Proteins; Proteomics; Radial; Rattus; Regenerative Medicine; Regulation; Role; STAT3 gene; Safety; Schwann Cells; Sensory; Signal Pathway; Site; Taste Buds; Testing; Therapeutic; Therapeutic Effect; Tissue Engineering; Tongue; Transplantation; Treatment Efficacy; Untranslated RNA; afferent nerve; base; cell motility; combinatorial; exosome; extracellular vesicles; immunoregulation; improved; intercellular communication; male; milk fat globule; nerve autograft; nerve injury; nerve stem cell; neurogenesis; novel; novel strategies; novel therapeutics; particle; peripheral nerve regeneration; pleiotropism; reconstruction; regenerative; regenerative therapy; repaired; response to injury; sciatic nerve; self-renewal; small hairpin RNA; standard care; stem cells; tissue regeneration

PROJECT SUMMARY/ABSTRACT Peripheral nerve injury (PNI) is a common and complex clinical challenge. Nerve autografts remain the “gold standard” for reconstruction of PNI with a large gap, but limited availability of donor nerves and several severe donor site morbidities significantly impede their clinic application. The combinatorial use of tissue engineered nerve guidance conduits (NGCs) with supportive cells and bioactive factors has the potential to be alternatives to nerve autografts. Mesenchymal stem cell (MSC)-based therapy has shown promises in regenerative medicine due to their multipotent, immunomodulatory/anti-inflammatory, and regenerative potentials. The therapeutic effects of MSCs most probably rely on their secretome, particularly the secretory extracellular vesicles (EVs) which play important roles in intercellular communication through transfer of various bioactive molecules e.g. lipids, proteins, and noncoding RNAs. Due to their comparable therapeutic efficacy to parental cells, MSC-EVs have been explored as novel cell-free therapeutics for a spectrum of diseases. However, the potential use of MSC-EVs in peripheral nerve regeneration remains largely unexplored. We have recently shown that human gingiva-derived MSCs (GMSCs) and their released EVs displayed comparable therapeutic effects on regeneration of taste bud/taste sensory nerves of rats and repair/regeneration of crush-injured sciatic nerves of mice possibly by promoting reprogramming of myelinated Schwann cells toward a repair phenotype. Our preliminary study showed that infusion of GMSC-EVs into the customized AxoGuard Nerve Connectors significantly improved their efficacy on regeneration of transected rat sciatic nerves. Through proteomic profiling, we identified a group of protein factors significantly enriched in GMSC-EVs, among which milk fat globule-epidermal growth factor-factor VIII (MFG-E8) is particularly interesting because of its pleiotropic biological functions. Our preliminary data also showed that MFG-E8 activated pSTAT3 and upregulated gene expressions involved in regulating repair phonotypic conversion of Schwann cells. We hypothesize that enriched MFG-E8 contributes a major role in GMSC-EVs mediated therapeutic effects on nerve regeneration by promoting the repair phenotypic conversion of Schwann cells. We proposed two specific aims to test the hypothesis: 1) determine the pleiotropic effects of MFG-E8 enriched in GMSC-EVs on the repair phenotypic conversion of Schwann cells; 2) investigate the critical role of MFG-E8 in GMSC EV-mediated therapeutic effects on regeneration of transected rat sciatic nerves. Our long-term goal is to develop a cell-free/stem cell- based product as a potential alternative to nerve autografts for regeneration of PNI with a large gap. Results from this study will lead us to submit an R01 grant application that will allow us to better understand the mechanisms underlying MSC-EV mediated therapeutic effects on nerve regeneration and to further explore the safety and efficacy of MSC-EVs in nerve regeneration in large animals and early human clinical trials.

项目摘要/摘要 周围神经损伤（PNI）是常见且复杂的临床挑战。神经自体移植仍然是“黄金 标准”用于重建PNI的差距很大，但供体神经的可用性有限，几个严重 供体现场的病毒严重阻碍了他们的诊所应用。组织工程的组合使用 具有支持性细胞和生物活性因子的神经引导导管（NGC）有可能成为替代方案 神经自体移植。间充质干细胞（MSC）基于基于的治疗已显示出再生的承诺 由于其多能，免疫调节/抗炎和再生潜力，药物。 MSC的治疗作用很可能依赖于其分泌型，尤其是分泌细胞外的 通过转移各种生物活性在细胞间交流中起重要作用的蔬菜（EV） 分子，例如脂质，蛋白质和非编码RNA。由于其可比的治疗效率与父母 细胞，MSC-EV已被探索为多种疾病的新型无细胞治疗。但是， MSC-EV在周围神经再生中的潜在使用仍然很大程度上是出乎意料的。我们最近有 显示人类牙龈衍生的MSC（GMSC）及其释放的电动汽车显示出可比的治疗 对大鼠的味蕾/味觉感官神经的再生的影响以及粉碎受伤的修复/再生 小鼠的坐骨神经通过促进髓鞘的雪旺细胞重新编程以修复 表型。我们的初步研究表明，将GMSC-evs输注到定制的Axoguard神经中 连接器可显着提高其对转移大鼠坐骨神经再生的效率。通过 蛋白质组学分析，我们确定了一组蛋白质因子在GMSC-EV中显着富集，其中 牛奶脂肪球 - 表皮生长因子因素VIII（MFG-E8）特别有趣，因为其多效性 生物功能。我们的初步数据还表明，MFG-E8激活了PSTAT3并更新了基因 涉及调节schwann细胞的修复语音转换的表达式。我们假设这一点 富集的MFG-E8在GMSC-EVS介导的治疗作用中对神经再生有重要作用 通过促进雪旺细胞的修复表型转化。我们提出了两个特定目标，以测试 假设：1）确定富含GMSC-EV的MFG-E8对修复表型的多效效应 雪旺细胞的转化； 2）研究MFG-E8在GMSC EV介导的疗法中的关键作用 对转移大鼠坐骨神经再生的影响。我们的长期目标是开发无细胞/干细胞 - 基于较大间隙的PNI再生的神经自体移植物的潜在替代品。结果 从这项研究中将导致我们提交R01赠款申请，这将使我们能够更好地了解 MSC-EV介导的治疗作用对神经再生的治疗作用，并进一步探索 MSC-EV在大型动物和早期人类临床试验中的神经再生中的安全性和效率。