Regulation of Tendon Repair with Stem Cell-Derived Extracellular Vesicles

干细胞来源的细胞外囊泡对肌腱修复的调节

• 批准号: 10929503
• 负责人: Hua Shen
• 金额: $ 50.15万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-18 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10929503
• 关键词: Affect; Anabolism; Biochemical; Biogenesis; Biological; Biological Assay; Cell Proliferation; Cell membrane; Cell physiology; Cells; Clinical; Collagen; Cumulative Trauma Disorders; Development; Disease; Economic Burden; Effectiveness; Equilibrium; Excision; Exhibits; Future; Generations; Goals; In Vitro; Individual; Infiltration; Inflammation; Inflammatory; Inflammatory Response; Injury; Knowledge; Ligaments; Macrophage; Mediating; Medical; Messenger RNA; Metabolism; Methods; MicroRNAs; Molecular; Molecular Target; Morphogenesis; Multivesicular Body; Mus; Natural regeneration; Operative Surgical Procedures; Orthopedics; Outcome; Pain; Parents; Pathway interactions; Patients; Persistent pain; Phenotype; Production; Proliferating; Proteomics; Quality of life; Recovery of Function; Regulation; Rehabilitation therapy; Signal Transduction; Sorting; Structure; Subgroup; Surface; TLR4 gene; Tendon Injuries; Tendon structure; Therapeutic; Time; Treatment Efficacy; United States; Vesicle; achilles tendon; adipose derived stem cell; clinical application; clinically relevant; cost; effectiveness evaluation; exosome; extracellular vesicles; gain of function; healing; improved; injury and repair; ligament injury; loss of function; microvesicles; overexpression; pre-clinical; regenerative; repair model; repaired; response; response to injury; scale up; side effect; stem cells; transcriptome sequencing

PROJECT SUMMARY/ABSTRACT Tendon and ligament injuries are among the most common and challenging orthopedic conditions. Many patients suffer from long-term pain and reduced function after injury. Substantial evidence indicates that these unsatisfactory outcomes result primarily from an imbalanced healing response with excessive and/or sustained inflammation and inadequate tendon regeneration. We discovered that extracellular vesicles produced by inflammation-primed adipose-derived stem cells (iEV) could potentially restore balanced healing by attenuating inflammation mediated by macrophages and stimulating regeneration driven by tendon cells. However, bulk iEV are heterogeneous. Differentially generated iEV carry different mRNA and microRNA molecules, which determine iEV functions. There is strong evidence that only a fraction of iEV carry the active molecules that balance the injury responses of macrophages and tendon cells, whereas many other iEV could potentially compromise tendon healing or cause unwanted side effects. Therefore, it is necessary to identify the iEV subpopulation and active cargo molecules specific for tendon repair and use them to develop disease-specific therapy to greatly enhance tendon repair. To achieve this goal, bulk iEV will be sorted into hierarchically organized subgroups based on vesicle generation and cargo sorting machineries that determine vesicle cargo composition. The subpopulation and active cargo molecules that specifically enhance tendon healing will be identified by their potency in regulating tendon cell and macrophage activities and functions. The therapeutic efficacy of identified iEV and cargo molecules for tendon repair will be examined in a preclinical mouse Achilles tendon injury and repair model. We anticipate that removal of functionally non-specific iEV or selective enrichment of functionally specific cargo molecules in bulk iEV will greatly increase the therapeutic efficacy and avoid potential adverse side effects of bulk iEV for tendon injury treatments. This study is a critical proof-of- concept analysis toward clinical application of extracellular vesicle (EV)-based therapy for tendon and ligament injuries, which will benefit millions of patients. This study also will discover the mechanism of iEV action for tendon injuries, and mechanisms underlying the molecular regulation of tendon cell and macrophage activities and functions. Knowledge gained about the active molecules, surface markers, and biogenesis of the iEV subpopulation specific for tendon repair will be used to increase the yield and release of these iEV by parent cells and scale up iEV production via immunoaffinity-based method. Our established approaches can be used for the future development of EV-based and disease-specific therapies for other medical conditions, which will improve the quality of life and outcomes of more patients.

项目概要/摘要 肌腱和韧带损伤是最常见和最具挑战性的骨科疾病之一。很多病人 受伤后遭受长期疼痛和功能下降。大量证据表明，这些 不令人满意的结果主要是由于过度和/或持续的治疗反应不平衡造成的 炎症和肌腱再生不足。我们发现细胞外囊泡是由 炎症引发的脂肪干细胞（iEV）可能通过减弱来恢复平衡愈合 由巨噬细胞介导的炎症和由肌腱细胞驱动的刺激再生。然而，批量iEV 是异质的。差异化产生的 iEV 携带不同的 mRNA 和 microRNA 分子， 确定 iEV 功能。有强有力的证据表明，只有一小部分 iEV 携带活性分子， 平衡巨噬细胞和肌腱细胞的损伤反应，而许多其他 iEV 可能 影响肌腱愈合或引起不良副作用。因此，有必要对iEV进行识别 肌腱修复特异性亚群和活性货物分子，并利用它们开发疾病特异性 疗法大大增强肌腱的修复。为了实现这一目标，批量iEV将分层分类 根据囊泡生成和确定囊泡货物的货物分类机器组织小组 作品。专门增强肌腱愈合的亚群和活性货物分子将是 通过其调节肌腱细胞和巨噬细胞活动和功能的效力来确定。治疗性的 已确定的 iEV 和货物分子用于肌腱修复的功效将在临床前小鼠跟腱中进行检查 肌腱损伤和修复模型。我们预计去除功能上非特异性的 iEV 或选择性 大量 iEV 中功能特异性货物分子的富集将大大提高治疗效果 避免批量 iEV 用于肌腱损伤治疗的潜在不良副作用。这项研究是一个重要的证明—— 基于细胞外囊泡（EV）的肌腱和韧带治疗临床应用的概念分析 伤，这将使数百万患者受益。这项研究还将发现 iEV 的作用机制 肌腱损伤以及肌腱细胞和巨噬细胞活动的分子调节机制 和功能。获得有关 iEV 活性分子、表面标记和生物发生的知识 肌腱修复特异的亚群将用于增加亲代这些 iEV 的产量和释放 细胞并通过基于免疫亲和力的方法扩大 iEV 生产。我们既定的方法可以使用 为未来开发针对其他医疗状况的基于 EV 的疾病特异性疗法，这将 改善更多患者的生活质量和治疗结果。