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: 10935538
• 负责人: Xianghong Luan
• 金额: $ 34.83万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-26 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10935538
• 关键词: New; craniofacial; bone; engineering; through

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.

血管影响骨生理的各个方面，包括矿物质生长 在新的骨形成期间。将骨头设计为血管化组织，我们现在有 专注于microRNA作为分子工程工具，因为它们的能力 可逆地针对多个监管网络。来自我们和其他的最新研究 实验室表明，miRNA促进骨谱系分化和 通过Wnt，BMP和Notch信号通路形成新的骨骼，也可以控制 骨骼分辨率。在我们寻求促进骨骼耦合骨骼的新颖疗法中 重塑和血管生成用于治疗大骨缺损的方法 将miR-27〜23〜24 microRNA簇确定为高度有希望的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'的未翻译区域（UTR）， 在血管生成抑制剂的UTR上与miR-24结合位点的联系 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，我们证明了miR-23-27-24群集成员 炎症组织中的表达大大降低。相反，mir-27模仿 患有严重垂直骨缺损的患病牙周组织的治疗导致 出乎意料的广泛的新骨形成，由80％的完整修复证明 垂直骨高和矿化组织的新沉积在大颅骨中 MiR-27应用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，新血管形成和 临界尺寸缺陷中大量的颅骨再生。在细胞上 水平，miR-27的过表达增加了酒精磷酸酶的活性和骨标记 基因表达，而抑制miR-27导致了急剧增加 骨质发生相关基因和骨骼分辨率的表达。 Mir-24和 miR-27促进了内皮细胞增殖和血管管的形成。 这些令人兴奋的初步数据促使我们提出了一种新颖的分子 可以利用的工程策略 miR-23/27/24群集成员与合适的脚手架和重新平衡 新的颅面骨形成的肺泡和颅骨稳态。