Nanolipidoids-Conjugated MicroRNA Enhance Oral and Cranial Bone Regeneration

纳米脂质结合的 MicroRNA 增强口腔和颅骨再生

• 批准号: 9106764
• 负责人: JAKE JINKUN CHEN
• 金额: $ 33万
• 依托单位: TUFTS UNIVERSITY BOSTON
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-03-01 至 2020-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9106764
• 关键词: Adverse effects; Anatomy; Animal Model; Animals; Biological; Biological Process; Bone Diseases; Bone Marrow; Bone Matrix; Bone Regeneration; Bone Tissue; Cells; Cephalic; Clinical; Clinical Treatment; Clinical Trials; Defect; Deformity; Drug Targeting; Engineering; Failure; Fracture; Fracture Healing; Future; Genes; Genome; Hepatitis C; Implant; In Situ; Individual; Joints; Label; Laboratories; Lead; Life; Malignant Neoplasms; Mammalian Cell; Mesenchymal Stem Cells; MicroRNAs; Molecular; Mus; Muscle; Natural regeneration; Nerve; Operative Surgical Procedures; Oral; Organ Transplantation; Osseointegration; Osteoblasts; Osteogenesis; Osteoporosis; Patients; Periodontitis; Phase; Physiological; Positioning Attribute; Procedures; Regenerative Medicine; Signal Transduction; Site; Stromal Cells; System; Testing; Therapeutic; Therapeutic Agents; Tissue Engineering; Tissues; Transcript; Transgenes; Transgenic Animals; Transgenic Mice; Translational Research; Transplantation; Untranslated RNA; Untranslated Regions; Viral Vector; Work; Wound Healing; adult stem cell; angiogenesis; base; beta catenin; bone; bone mass; clinical application; cost; craniofacial; experience; human disease; in vivo; induced pluripotent stem cell; innovation; interest; mouse genome; nanomaterials; nanoparticle; novel; organ growth; osteogenic; osteogenic protein; overexpression; physical property; preclinical trial; promoter; public health relevance; scaffold; skills; stem cell biology; stem cell differentiation; success; therapeutic miRNA; therapeutic target; tissue regeneration; transcription factor; translational study; wound

 DESCRIPTION (provided by applicant): While over 500,000 surgeries correcting bone deformities and critical size defects occur each year in the US, 50% of the procedures end in failure. Therefore, using molecular engineering approaches to regenerate lost bone with normal anatomic contour, physical properties and biological function is of enormous importance. MicroRNAs (miRNAs) are non-coding RNAs that are emerging as new and powerful drug targets. Currently there are 157 preclinical and clinical trials of miRNA-based therapeutics. We have identified a novel miRNA, miR-335-5p, which targets the 3'-UTR sequence of DKK1, a Wnt signal antagonist, and down-regulates DKK1 expression. We also revealed that miR-335-5p is an important regulator for osteogenic differentiation. We have further generated a transgenic mouse line in which miR-335-5p gene is overexpressed in bone tissue showing enhanced expression of marker genes and increased bone mass accrual. Based on these interesting findings we plan to use miR-335-5p as a therapeutic target to enhance oral and craniofacial bone regeneration using our newly synthesized and characterized lipidoid-miRNA nanoparticles as a novel delivery vehicle. Aim 1. We will determine regulatory and osteogenic effects of miR-335-5p overexpression in bone regeneration and wound repairing using a transgenic animal model. Our hypothesis is that since miR-335-5p transgene is incorporated into mouse genome its super-physical transcript levels will boost the bone forming capacity in target cells. Aim 2. We will develop two therapeutic strategies with a nanoparticle lipidoid system to deliver miR-335-5p molecules in vivo for oral and craniofacial bone regeneration. Firstly, we will transplant miR-335-5p-modified bone marrow stromal cells into bone wound site to enhance bone formation. Our hypothesis is that miR-335-5p specifically suppresses DKK1 and activates Wnt/β−Catenin signals, which further triggers an osteogenic cascade leading BMSC differentiation towards osteoblastic lineage and an increased bone formation capability. Secondly, we will determine the effect of miR-335-5p in promoting bone-forming cells in situ and enhancing wound healing and tissue regeneration. Our hypothesis is that local administration of miR-335-5p in wound site directly leads mesenchymal stem cells differentiation into bone forming cells, accelerating wound healing and bone regeneration. This translational research will explore the potential of miR-335-5p as a therapeutic agent for promoting bone regeneration and wound healing. This innovative miRNA-based therapy has potential for the clinical treatment of other bone disorders including osteoporosis, fracture, periodontitis and impaired osseointegration of bone/joint implants.

 描述（由申请人提供）：虽然美国每年发生超过 500,000 例矫正骨骼畸形和临界尺寸缺陷的手术，但其中 50% 的手术以失败告终。因此，利用分子工程方法来再生具有正常解剖轮廓、物理特性和生物功能的丢失骨具有非常重要的意义。 MicroRNA (miRNA) 是非编码 RNA，正在成为新的强大药物靶点。目前有 157 项基于 miRNA 的疗法的临床前和临床试验。我们发现了一种新的 miRNA，miR-335-5p，它靶向 Wnt 信号拮抗剂 DKK1 的 3'-UTR 序列，并下调 DKK1 的表达。我们还发现 miR-335-5p 是成骨分化的重要调节因子。我们进一步生成了转基因小鼠品系，其中 miR-335-5p 基因在骨组织中过度表达，显示出标记基因表达增强和骨量增加增加。基于这些有趣的发现，我们计划使用 miR-335-5p 作为治疗靶点，使用我们新合成和表征的类脂质-miRNA 纳米颗粒作为新型递送载体来增强口腔和颅面骨再生。 目标 1. 我们将使用转基因动物模型确定 miR-335-5p 过表达在骨再生和伤口修复中的调节和成骨作用。我们的假设是，由于 miR-335-5p 转基因被整合到小鼠基因组中，其超物理转录水平将提高靶细胞的骨形成能力。 目标 2. 我们将开发两种治疗策略，利用纳米颗粒类脂质系统在体内递送 miR-335-5p 分子，用于口腔和颅面骨再生。 首先，我们将miR-335-5p修饰的骨髓基质细胞移植到骨伤口部位以增强骨形成。我们的假设是，miR-335-5p 特异性抑制 DKK1 并激活 Wnt/β−Catenin 信号，进一步触发成骨级联反应，导致 BMSC 向成骨细胞谱系分化并增加骨形成能力。 其次，我们将确定miR-335-5p在促进原位骨形成细胞以及增强伤口愈合和组织再生方面的作用。我们的假设是，在伤口部位局部施用 miR-335-5p 直接导致间充质干细胞分化为骨形成细胞，加速伤口愈合和骨再生。 这项转化研究将探索 miR-335-5p 作为促进骨再生和伤口愈合的治疗剂的潜力。这种基于 miRNA 的创新疗法具有临床治疗其他骨骼疾病的潜力，包括骨质疏松症、骨折、牙周炎和骨/关节植入物骨整合受损。