Extracellular Matrix Vesicles and Intercellular Communication of Osteoblast-lineage Cells

成骨细胞谱系细胞的细胞外基质囊泡和细胞间通讯

• 批准号: 10679255
• 负责人: Anne Michelle Skelton
• 金额: $ 4.43万
• 依托单位: VIRGINIA COMMONWEALTH UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-06-01 至 2027-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10679255
• 关键词: Adult; Alkaline Phosphatase; Autologous; Binding; Biological; Biological Assay; Bone Development; Bone Marrow; Bone Regeneration; Bone Transplantation; Cartilage; Cartilage Matrix; Cell Lineage; Cells; Characteristics; Chemicals; Chondrocytes; Chronic; Coculture Techniques; Collagen Fibril; Collagen Type I; Culture Media; Defect; Deposition; Differentiation Antigens; Embryo; Enzymes; Epiphysial cartilage; Experimental Models; Extracellular Matrix; Femur; Foundations; Fracture; Goals; Human body; Hybrid Cells; Hydrogels; Hydroxyapatites; In Vitro; Injectable; Integrin Binding; Lipids; Literature; Luciferases; Mature Bone; Measures; Mediating; Mediation; Membrane; MicroRNAs; Minerals; Modeling; Morphology; Mus; Nucleic Acids; Operative Surgical Procedures; Organelles; Orthopedics; Osteoblasts; Osteocalcin; Osteogenesis; Pathology; Pathway interactions; Patients; Pattern; Phenotype; Phospholipids; Physiologic Ossification; Physiologic calcification; Play; Population; Population Heterogeneity; Process; Proliferating; Property; Proteins; Regenerative Medicine; Research; Role; Signal Pathway; Signal Transduction; Signaling Molecule; Site; Stromal Cells; System; Testing; Therapeutic; Tissues; Translating; Transmission Electron Microscopy; Trypsin; Untranslated RNA; Vesicle; Western Blotting; body system; bone; bone fracture repair; bone healing; bone repair; calcification; calcium phosphate; exosome; extracellular vesicles; fetal; healing; improved; in vivo; in vivo Model; intercellular communication; long bone; mRNA Expression; microCT; mineralization; nanoparticle; nanosized; novel therapeutics; osteoblast differentiation; osteogenic; protein expression; regenerative therapy; repaired; response; standard care; success; therapeutic evaluation; vesicular release

PROJECT SUMMARY Autologous bone grafting, the current gold standard for treating critical sized bone defects and chronic nonunions, has limited success, leaving a need for adjunct treatments to contribute to overall bone healing. Extracellular vesicles, of which there are many types, are a promising new cell-free, membrane-bound therapeutic in the field of regenerative medicine. Specific to the field of bone regeneration and repair are matrix vesicles; small extracellular vesicles released by mineralization-competent cells that become anchored in the extracellular matrix and are a necessary component of mineralized bone formation via endochondral ossification. They are involved in cellular signaling via small, non-coding microRNA in the growth plate, suggesting that they may play a similar role in bone. Previous research indicates that chondrocyte-derived matrix vesicles are enriched with microRNA and other cell signaling molecules that contribute to their ability to influence proliferation and differentiation of target cells, however the role and mechanism of action of osteoblast-derived matrix vesicles, particularly in bone as opposed to the growth plate, remains to be elucidated. With this information, the biological mediation of bone development and regeneration that matrix vesicles provide may translate into novel therapeutic potential for orthopedic pathologies, including critical size bone defects and fracture nonunions. Therefore, we hypothesize that matrix vesicles produced by osteoblast-lineage cells, as a specific subset of extracellular vesicle, use their microRNA cargo to enhance osteogenic differentiation and proliferation, drive osteogenesis, and aid in bone defect healing. First, we aim to determine the relationship between osteoblast- derived matrix vesicles and another class of extracellular vesicle, the exosome. Next, we aim to determine the pathways targeted by matrix vesicle microRNA cargo. Finally, we aim to determine the extent to which osteoblast-derived matrix vesicles modulate osteogenesis and bone defect healing. To test these aims, we will use osteoblast-lineage cells to derive matrix vesicles and characterize protein expression, size, and morphology. We will also use in vitro experimental models to assess pathway involvement and evaluate co-culture response with osteoblast-like cells. To test the therapeutic potential, we will use a translatable in vivo model of a mouse long bone defect and a biorthogonal injectable hydrogel to deliver matrix vesicles. We expect to find that osteoblast-derived matrix vesicles are a specialized subclass of extracellular vesicle with microRNA cargo that targets the canonical Wnt pathway to activate cellular signaling and leads to increased osteoblastic differentiation of target cells when co-cultured in vitro. Our in vivo model is expected to demonstrate improved healing upon treatment with hydrogel-delivered matrix vesicles. All of which together demonstrates the role matrix vesicles play in the coordinated effort to form new bone and their viability as a therapeutic option to improve bone healing.

项目概要 自体骨移植是目前治疗严重骨缺损和慢性骨缺损的金标准 骨不连的成功率有限，因此需要辅助治疗来促进整体骨愈合。 细胞外囊泡有多种类型，是一种有前途的新型无细胞、膜结合的囊泡 再生医学领域的治疗。专门针对骨再生和修复领域的是基质 囊泡；由矿化活性细胞释放的小细胞外囊泡，锚定在 细胞外基质，是通过软骨内骨化形成矿化骨的必要成分。 它们通过生长板中的小非编码 microRNA 参与细胞信号传导，这表明它们 可能在骨骼中发挥类似的作用。先前的研究表明软骨细胞衍生的基质囊泡 富含 microRNA 和其他细胞信号分子，有助于影响增殖 和靶细胞的分化，然而成骨细胞衍生基质的作用和作用机制 囊泡，特别是在骨中而不是生长板中的囊泡，仍有待阐明。有了这些信息， 基质囊泡提供的骨发育和再生的生物介导可能转化为新的 骨科病理的治疗潜力，包括临界尺寸骨缺损和骨折不愈合。 因此，我们假设由成骨细胞谱系细胞产生的基质囊泡是 细胞外囊泡，利用其 microRNA 货物增强成骨分化和增殖，驱动 成骨，并有助于骨缺损愈合。首先，我们的目的是确定成骨细胞之间的关系 衍生的基质囊泡和另一类细胞外囊泡，外泌体。接下来，我们的目标是确定 基质囊泡 microRNA 货物靶向的途径。最后，我们的目标是确定 成骨细胞衍生的基质囊泡调节成骨和骨缺损愈合。为了测试这些目标，我们将 使用成骨细胞谱系细胞衍生基质囊泡并表征蛋白质表达、大小和形态。 我们还将使用体外实验模型来评估通路参与并评估共培养反应 与成骨细胞样细胞。为了测试治疗潜力，我们将使用可翻译的小鼠体内模型 长骨缺损和双正交可注射水凝胶以输送基质囊泡。我们期望发现 成骨细胞衍生的基质囊泡是细胞外囊泡的一个特殊亚类，其含有 microRNA 靶向经典 Wnt 通路激活细胞信号传导并导致成骨细胞分化增加 体外共培养时的靶细胞。我们的体内模型预计将展示出改善的愈合效果 用水凝胶递送的基质囊泡进行治疗。所有这些共同证明了基质囊泡的作用 协调努力形成新骨及其作为改善骨愈合的治疗选择的可行性。