New craniofacial bone engineering through miR-23-27-24 cluster mediated osteogenic-angiogenic coupling

通过miR-23-27-24簇介导的成骨-血管生成耦合的新颅面骨工程

• 批准号: 10252923
• 负责人: Xianghong Luan
• 金额: $ 32.49万
• 依托单位: TEXAS A&M UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-03 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10252923
• 关键词: Affect; Alkaline Phosphatase; Alveolar; American; Angiogenesis Inhibitors; Animal Model; Animals; BCL2L11 gene; Binding Sites; Biomedical Engineering; Blood Vessels; Bone Marrow; Bone Regeneration; Bone Resorption; Bone remodeling; Calvaria; Cell Culture Techniques; Cell Proliferation; Coupled; Coupling; Craniofacial Abnormalities; Data; Defect; Deposition; Endothelial Cells; Engineering; Equilibrium; Family; Gene Expression; Genes; Growth; Health; Health Benefit; Height; Homeostasis; Implant; In Vitro; Inflammation; Inflammatory; Laboratories; Mediating; Methodology; MicroRNAs; Minerals; Molecular; Natural regeneration; Notch Signaling Pathway; Oral; Osteogenesis; Periodontal Diseases; Periodontitis; Physiology; Reagent; Role; Small RNA; Testing; Tissue Engineering; Tissues; Tube; Untranslated Regions; Vascularization; angiogenesis; base; blood vessel development; bone; bone cell; bone engineering; bone loss; controlled release; craniofacial bone; experimental study; implantation; improved; in vitro Model; in vivo; member; monocyte; nanoparticle; novel; novel therapeutics; osteoclastogenesis; osteogenic; overexpression; progenitor; restoration; scaffold; subcutaneous; therapeutic miRNA; tool

Blood vessels affect all aspects of bone physiology, including mineral growth during new bone formation. To engineer bone as a vascularized tissue we have now focused on microRNAs as molecular engineering tools because of their ability to reversibly target multiple regulatory networks. Recent studies from our and other laboratories have demonstrated that miRNAs promote bone lineage differentiation and new bone formation through WNT, BMP, and Notch signaling pathways and also control bone resorption. In our quest for novel therapeutics that promote coupled bone remodeling and angiogenesis for the treatment of large bone defects we have here identified the miR-27~23~24 microRNA cluster as a highly promising miRNA candidate reagent. Our preliminary analysis revealed multiple miR-23 and miR-27 binding sites on the untranslated region (UTR) of the Wnt antagonist Sfrp1 and Nlk 3’, which in conjunction with miR-24 binding sites on the UTR of the angiogenesis inhibitors BIM and SEMA4A demonstrate the potential of the miR-27-23-24 microRNA cluster for coupled osteogenic and angiogenic molecular engineering strategies. In preliminary studies, we have demonstrated that miR-23-27-24 cluster member expression was greatly reduced in inflammatory tissues. Conversely, miR-27 mimic treatment of diseased periodontal tissues with severe vertical bone defects resulted in unexpectedly extensive new bone formation as evidenced by 80% complete restoration of vertical bone height and new deposition of mineralized tissue in large calvarial bone defects after 8 weeks of miR-27 application. Implants containing PEG-PLGA-PLL-miR- 27 nanoparticles promoted matrix remodeling, new blood vessel formation, and substantial amounts of calvarial bone regeneration in critical size defects. On a cellular level, miR-27 overexpression increased alkaline phosphatase activity and bone marker gene expression, while inhibition of miR-27 resulted in a dramatic increase in the expression of osteoclastogenesis related genes and bone resorption. Both miR-24 and miR-27 promoted endothelial cell proliferation and blood vessel tube formation. Together, these exciting preliminary data prompted us to propose a novel molecular engineering strategy that will harness the osteogenic and pro-angiogenic qualities of miR-23/27/24 cluster members in conjunction with suitable scaffolds and re-balance alveolar and calvarial bone homeostasis for new craniofacial bone formation.

血管影响骨骼生理学的各个方面，包括矿物质生长 在新骨形成过程中。为了将骨骼设计为血管化组织，我们现在拥有 专注于将 microRNA 作为分子工程工具，因为它们能够 可逆地针对多个监管网络。我们和其他人的最新研究 实验室已经证明 miRNA 促进骨谱系分化 通过 WNT、BMP 和 Notch 信号通路控制新骨形成 骨吸收。在我们寻求促进骨耦合的新疗法的过程中 我们这里有用于治疗大骨缺损的重塑和血管生成 鉴定出 miR-27~23~24 microRNA 簇是非常有前途的 miRNA 候选者 试剂。我们的初步分析揭示了多个 miR-23 和 miR-27 结合位点 Wnt 拮抗剂 Sfrp1 和 Nlk 3' 的非翻译区 (UTR)，其中 与血管生成抑制剂 BIM 的 UTR 上的 miR-24 结合位点结合 和 SEMA4A 证明了 miR-27-23-24 microRNA 簇的潜力 耦合成骨和血管生成分子工程策略。 在初步研究中，我们已经证明 miR-23-27-24 簇成员 炎症组织中的表达大大降低。相反，miR-27 模拟 治疗具有严重垂直骨缺损的患病牙周组织导致 80% 的完全修复证明出乎意料地广泛的新骨形成 大颅骨中垂直骨高度的变化和矿化组织的新沉积 应用 miR-27 8 周后出现缺陷。含有 PEG-PLGA-PLL-miR- 的植入物 27 纳米颗粒促进基质重塑、新血管形成， 临界尺寸缺损处有大量颅骨再生。在蜂窝网络上 水平，miR-27 过表达增加碱性磷酸酶活性和骨标志物 基因表达，而抑制 miR-27 导致 破骨细胞生成相关基因和骨吸收的表达。 miR-24 和 miR-27促进内皮细胞增殖和血管形成。 总之，这些令人兴奋的初步数据促使我们提出了一种新的分子 工程策略将利用成骨和促血管生成的特性 miR-23/27/24簇成员与合适的支架结合并重新平衡 牙槽骨和颅骨骨稳态，促进新颅面骨的形成。