Oral and Craniofacial Bone Regeneration using MicroRNA Modulation

使用 MicroRNA 调制进行口腔和颅面骨再生

• 批准号: 10192700
• 负责人: BRAD A AMENDT
• 金额: $ 36.22万
• 依托单位: UNIVERSITY OF IOWA
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-07-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10192700
• 关键词: Affect; Allografting; Autologous Transplantation; Bioinformatics; Bone Development; Bone Regeneration; Bone Transplantation; Calvaria; Collagen; Craniofacial Abnormalities; DNA; Data; Defect; Effectiveness; Family; Gene Expression Regulation; Genetic Transcription; Glycolates; Goals; Human; In Vitro; Inflammation; Mediating; Mesenchymal Stem Cells; MicroRNAs; Modeling; Molecular; Morbidity - disease rate; NF-kappa B; Non-Viral Vector; Oral; Osteogenesis; Pathway interactions; Periodontitis; Physiological; Plasmids; Porifera; Publishing; Rattus; Regenerative capacity; Regulation; Reporter Genes; Repression; Role; Signal Pathway; Signal Transduction; Site; System; Testing; Tissue Engineering; Transfection; Untranslated RNA; WNT Signaling Pathway; base; beta catenin; biomaterial compatibility; bone; bone marrow mesenchymal stem cell; craniofacial; craniofacial bone; cytokine; disease transmission; disorder risk; gene delivery system; gene therapy; improved; in vivo; in vivo regeneration; inhibitor/antagonist; member; nanoparticle; novel therapeutics; osteogenic; overexpression; programs; standard care; therapeutic miRNA; transcription factor; transcriptome sequencing

Project Summary Autografts and allografts are current standard strategies for bone repair of oral and craniofacial defects. However, each possesses limitations, such as donor-site morbidity with the use of autograft and the risk of dis- ease transmission with the use of allograft. Synthetic bone-graft substitute based on a tissue engineering strat- egy has represented an alternative approach to overcome these inherent limitations. MicroRNAs (miRs) are small non-coding RNAs that have emerged as important transcriptional regulators in both physiologic and pathophysiological conditions. We have previously shown that members of the miR-200 family actively regulate osteogenic differentiation, bone development, and inflammation. Our long-term goal is to develop a miR-based gene therapy program that can be used to effectively promote osteogenic differentiation and bone regeneration for restoring oral and craniofacial bone defects. Our recent studies showed that miR-200c loaded in collagen sponge promoted bone formation in vivo. We also observed that miR-200c up-regulated the activity of Wnt sig- naling and reduced Sox2 and Klf4. Relatedly, miR-200a has been demonstrated to directly suppress Wnt and BMP/TGF signal pathways. In our preliminary studies we have shown that inhibiting miR-200a using our Plas- mid-based miR Inhibitor System (PMIS) can effectively improve osteogenic differentiation and bone formation in vivo. Our objectives in this application are to understand and optimize the molecular function of miR-200c and PMIS-200a (miR-200a inhibitor) on osteogenic differentiation and bone formation and to validate that their local administration can be used to improve bone regeneration. The central hypothesis of this project is that miR-200c combined with miR-200a inhibition improve osteogenic differentiation by up-regulating Wnt and TGF- /BMP signaling. Plasmid miR-200c and PMIS-200a delivery can be optimized to promote bone regeneration for oral and craniofacial defects. In this project, we will determine the molecular function of miR-200c on Wnt sig- naling and the roles of Sox2 and Klf4 in the osteogenic differentiation mediated by miR-200c (Aim 1). We will also determine the function of PMIS-200a on osteogenic differentiation and bone formation by understanding the regulation of PMIS-200a on Wnt and BMP/TGF signaling (Aim 2). We will improve the effectiveness of miR-200c and PMIS-200a by optimizing a gene delivery system using biodegradable nanoparticles. In the Aim 3 we will use critical-sized calvarial defects and periodontitis-induced bone defects in rat models to investigate whether miR-200c combined with PMIS-200a can induce significant bone regeneration in vivo. At the comple- tion of this project, we will have significantly expanded our understanding of the molecular function of miR-200c and miR-200a inhibition and demonstrated the translational capabilities of these miRs for oral and craniofacial bone regeneration.

项目摘要 自体骨和同种异体骨是目前口腔和颅面骨缺损修复的标准策略。 然而，每一种方法都有局限性，例如使用自体移植物的供体部位发病率和手术后并发症的风险。 通过同种异体移植减轻传染。基于组织工程的人工骨替代物 egy代表了克服这些固有局限性的另一种方法。microRNAs（miRs） 小的非编码RNA已成为重要的转录调节因子，在生理和 病理生理条件。我们之前已经证明miR-200家族的成员积极调节 成骨分化、骨发育和炎症。我们的长期目标是开发一种基于miR的 可用于有效促进成骨分化和骨再生的基因治疗程序 用于修复口腔和颅面骨缺损。我们最近的研究表明，胶原蛋白中的miR-200 c 海绵促进体内骨形成。我们还观察到，miR-200 c上调了Wnt sig-1的活性。 使Sox 2和Klf 4减少。相关地，已经证明miR-200 a直接抑制Wnt和 BMP/TGF信号通路。在我们的初步研究中，我们已经表明，使用我们的Plas抑制miR-200 a， 中基miR抑制剂系统（PMIS）可有效促进成骨分化和骨形成 in vivo.本申请的目的是了解和优化miR-200 c的分子功能 和PMIS-200 a（miR-200 a抑制剂）对成骨分化和骨形成的影响，并验证它们在成骨分化和骨形成中的作用。 局部给药可用于改善骨再生。这个项目的核心假设是， miR-200 c联合miR-200 a抑制通过上调Wnt和TGF-β 1促进成骨分化 /BMP信号。质粒miR-200 c和PMIS-200 a的递送可以被优化以促进骨再生， 口腔和颅面缺陷。在这个项目中，我们将确定miR-200 c在Wnt信号通路中的分子功能。 naling以及Sox 2和Klf 4在miR-200 c介导的成骨分化中的作用（Aim 1）。我们将 通过了解PMIS-200 a在成骨分化和骨形成中的作用， PMIS-200 a对Wnt和BMP/TGF信号通路的调节（目的2）。我们将提高 miR-200 c和PMIS-200 a的基因表达。在Aim 3我们将使用大鼠颅骨缺损和牙周炎引起的骨缺损模型来研究 miR-200 c与PMIS-200 a联合是否能在体内诱导显著的骨再生。在完成- 通过这个项目，我们将大大扩展我们对miR-200 c分子功能的理解 和miR-200 a抑制，并证明了这些miR对口腔和颅面的翻译能力。 骨再生

项目成果

microRNA-126 inhibits vascular cell adhesion molecule-1 and interleukin-1beta in human dental pulp cells.

• DOI: 10.1002/jcla.24371
• 发表时间: 2022-05
• 期刊: Journal of clinical laboratory analysis
• 影响因子: 2.7

The miR-200 family is required for ectodermal organ development through the regulation of the epithelial stem cell niche.

• DOI: 10.1002/stem.3342
• 发表时间: 2021-06
• 期刊: Stem cells (Dayton, Ohio)
• 作者: Sweat M;Sweat Y;Yu W;Su D;Leonard RJ;Eliason SL;Amendt BA
• 通讯作者: Amendt BA

Six2 regulates Pax9 expression, palatogenesis and craniofacial bone formation.

• DOI: 10.1016/j.ydbio.2019.11.010
• 发表时间: 2020-02-15
• 期刊: Developmental biology
• 影响因子: 2.7
• 作者: Sweat YY;Sweat M;Mansaray M;Cao H;Eliason S;Adeyemo WL;Gowans LJJ;Eshete MA;Anand D;Chalkley C;Saadi I;Lachke SA;Butali A;Amendt BA
• 通讯作者: Amendt BA