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: 10180941
• 负责人: Anh D Le
• 金额: $ 24.38万
• 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 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10180941
• 关键词: Animals; Anti-Inflammatory Agents; Apoptotic; Applications Grants; Autologous Transplantation; Biological Process; 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 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 repair; nerve stem cell; neurogenesis; novel; novel strategies; novel therapeutics; particle; peripheral nerve regeneration; peripheral nerve repair; pleiotropism; reconstruction; regeneration potential; regenerative therapy; repaired; response to injury; sciatic nerve; sciatic nerve injury; self-renewal; small hairpin RNA; standard care; stem cell therapy; 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.

项目概要/摘要 周围神经损伤（PNI）是一种常见且复杂的临床挑战。自体神经移植仍然是“黄金” 标准”用于重建具有较大间隙的 PNI，但供体神经的可用性有限，并且存在一些严重的问题 供体部位的发病率严重阻碍了其临床应用。组织工程的组合应用 具有支持细胞和生物活性因子的神经引导导管（NGC）有可能成为替代品 自体神经移植物。基于间充质干细胞（MSC）的疗法在再生方面显示出希望 由于其多能、免疫调节/抗炎和再生潜力而成为医学领域。这 间充质干细胞的治疗效果很可能依赖于它们的分泌组，特别是分泌性细胞外 囊泡（EV）通过转移各种生物活性在细胞间通讯中发挥重要作用 分子，例如脂质、蛋白质和非编码 RNA。由于其治疗效果与父母相当 细胞，MSC-EV 已被探索作为治疗一系列疾病的新型无细胞疗法。然而， MSC-EV 在周围神经再生中的潜在用途在很大程度上仍未被探索。我们最近有 研究表明，人牙龈来源的间充质干细胞 (GMSC) 及其释放的 EV 具有相当的治疗效果 对大鼠味蕾/味觉神经再生及挤压损伤修复/再生的影响 小鼠坐骨神经可能通过促进有髓雪旺细胞重编程修复 表型。我们的初步研究表明，将 GMSC-EV 注入定制的 AxoGuard Nerve 连接器显着提高了横断大鼠坐骨神经再生的功效。通过 通过蛋白质组分析，我们鉴定了一组在 GMSC-EV 中显着富集的蛋白质因子，其中 乳脂肪球-表皮生长因子-因子 VIII (MFG-E8) 由于其多效性而特别令人感兴趣 生物学功能。我们的初步数据还表明MFG-E8激活pSTAT3并上调基因 参与调节雪旺细胞修复表型转换的表达。我们假设 富集的 MFG-E8 在 GMSC-EV 介导的神经再生治疗作用中发挥重要作用 通过促进雪旺细胞的修复表型转换。我们提出了两个具体目标来测试 假设：1）确定富含GMSC-EV的MFG-E8对修复表型的多效性作用 雪旺细胞的转化； 2) 研究MFG-E8在GMSC EV介导的治疗中的关键作用 对横断大鼠坐骨神经再生的影响。我们的长期目标是开发一种无细胞/干细胞- 基于产品作为神经自体移植的潜在替代品，用于具有较大间隙的 PNI 再生。结果 这项研究将引导我们提交 R01 拨款申请，这将使我们能够更好地了解 MSC-EV 介导的神经再生治疗作用的机制，并进一步探讨 MSC-EV 在大型动物神经再生和早期人体临床试验中的安全性和有效性。

项目成果

Gingiva-Derived Mesenchymal Stem Cells: Potential Application in Tissue Engineering and Regenerative Medicine - A Comprehensive Review.

• DOI: 10.3389/fimmu.2021.667221
• 发表时间: 2021
• 期刊: Frontiers in immunology
• 影响因子: 7.3
• 作者: Kim D;Lee AE;Xu Q;Zhang Q;Le AD
• 通讯作者: Le AD

Implantation of a nerve protector embedded with human GMSC-derived Schwann-like cells accelerates regeneration of crush-injured rat sciatic nerves.

• DOI: 10.1186/s13287-022-02947-4
• 发表时间: 2022-06-20
• 期刊: Stem cell research & therapy
• 影响因子: 7.5

Harnessing 3D collagen hydrogel-directed conversion of human GMSCs into SCP-like cells to generate functionalized nerve conduits.

• DOI: 10.1038/s41536-021-00170-y
• 发表时间: 2021-09-30
• 期刊: NPJ Regenerative medicine
• 影响因子: 7.2
• 作者: Zhang Q;Nguyen P;Burrell JC;Zeng J;Shi S;Shanti RM;Kulischak G;Cullen DK;Le AD
• 通讯作者: Le AD