Roles of Noncoding RNA in Bone Regeneration

此续订赠款申请代表了当前赠款(DE25681)重点的深化扩展 关于microRNA(MiR)-335-5P的骨增强作用，这是我们实验室首次鉴定和表征的。我们 发表同行评议论文9余篇，并多次获奖报道成果。我们有 最近报道了我们新发现的miR-335-5p抑制破骨细胞分化和骨的功能 再吸收。我们进一步发现miR-335-5P是通过与3‘非编码区元件结合而发挥抑制作用的 IGSF3(免疫球蛋白超家族，成员3)。Igsf3基因S在破骨细胞形成过程中的表达上调 分化程度与miR-335-5p表达呈负相关。这一新发现与 MiR-335-5p特有的合成代谢成骨作用，使我们推测miR-335-5p是一种有效的 治疗骨质疏松症及其相关骨病的候选药物，其中 骨形成和吸收受到干扰。目前，骨质疏松症的治疗包括抗吸收药物。 和合成代谢成骨药物。然而，这些药物要么针对骨吸收，要么针对骨形成。 路径，但不是两条都有。许多以蛋白质为基础的疗法都有副作用和较高的 成本。我们已经产生了miR-335-5P基因敲除(功能丧失)和过度表达 (功能获得)小鼠。这两个鼠标系列将提供最先进和最复杂的方法 为了达到我们的研究目的进行基因操作。与科学家和生物工程师在 新泽西理工学院，我们已经开发出新型和尖端的靶向纳米颗粒 首次将miR-335-5p精确地输送到靶细胞，在两种细胞中都能发挥其双重作用 骨吸收和骨形成途径。目的1.探讨新陈代谢综合征的分子机制。 发现miR-335-5p在抑制破骨细胞活性和骨吸收方面的作用；目的2.使用我们的 新建立的miR-335-5p基因敲除和过表达小鼠研究其多层功能 研究miR-335-5p在骨代谢中的作用；目的3.应用新开发的靶向纳米粒给药 MIR-335-5P对特定细胞类型的作用，并确定其对骨创伤愈合和逆转的治疗作用 骨质疏松。我们将确定miR-335-5p的治疗效果，给出有效浓度，最佳 频率和准确的给药持续时间，以最大限度地发挥其在细胞和组织中的功能 级别。这个更新项目在概念、技术和干预方面都是创新的。有优势的 基于miRNA的治疗的特点将使这项翻译研究改变理解的范式， 治疗并最终治愈骨质疏松症及其相关的骨病。一支跨学科的团队 具有互补和协同技能的研究人员将进行研究(杰克·陈-实验 病理与骨生物学；启胜屠呦呦细胞与分子生物学；小阳徐-生物材料与药物 交付)。

This renewal grant application represents a deepened extension of the current grant (DE25681) focusing on the bone-enhancing effects of microRNA (miR)-335-5p, first identified and characterized in our laboratory. We have published over 9 peer-reviewed papers and won many prizes and awards in reporting the results. We have recently reported our new discovery of the function of miR-335-5p in inhibiting osteoclast differentiation and bone resorption. We further found that miR-335-5p exerts its inhibitory effect through its binding to the 3’UTR elements of igsf3 (immunoglobulin superfamily, member 3). Igsf3’s increased expression during the process of osteoclast differentiation is reversely correlated with the expression of miR-335-5p. This new discovery together with the well-characterized anabolic osteogenic effect of miR-335-5p, has led us to speculate that miR-335-5p is a potent pharmaceutical candidate for treating osteoporosis and its related bone disorders, where the balance between bone formation and resorption is disturbed. At present, osteoporosis treatments include anti-resorptive drugs and anabolic bone-forming drugs. However, these drugs target either the bone-resorption or bone-formation pathway, but not both. Many protein-based therapies have the disadvantages including side-effects and the high cost. We have generated both miR-335-5p gene knockout (loss-of-function) and overexpression (gain-of-function) mice. These two mouse lines will provide the most advanced and sophisticated approaches for gene manipulation to achieve our research purposes. Collaborating with scientists and bioengineers at the New Jersey Institute of Technology, we have developed novel and cutting-edge targeted nanoparticles for the first time to precisely deliver miR-335-5p to the target cells where it can exert its dual-effects in both bone-resorption and bone-formation pahways. Aim 1. To explore the molecular mechanism of the newly discovered function of miR-335-5p in suppressing osteoclast activity and bone resorption; Aim 2. To use our newly generated miR-335-5p gene knockout and overexpression mice to characterize the multilayered functions of miR-335-5p in bone metabolism; Aim 3. To apply newly developed targeted nanoparticles to deliver miR-335-5p to specific cell types and determine its therapeutic effects on bone wound healing and reversal of osteoporosis. We will determine the therapeutic effects of miR-335-5p given effective concentration, optimal frequency, and accurate duration of administration to maximize its functions at both cellular and organismal levels. This renewal project is conceptually, technically, and interventionally innovative. The advantageous features of miRNA-based therapy will allow this translational study to shift the paradigm in understanding, treating and ultimately curing osteoporosis and its related bone disorders. An interdisciplinary team of investigators with complementary and synergistic skills will conduct the studies (Jake Chen – experimental pathology and bone biology; Qisheng Tu – cell and molecular biology; Xiaoyang Xu – biomaterials and drug delivery).