Roles of Noncoding RNA in Bone Regeneration

此更新补助金申请代表了当前补助金（DE 25681）的深化扩展， 对microRNA（miR）-335-5p的骨增强作用的研究，这是我们实验室首次发现和表征的。我们 发表了超过9篇同行评审论文，并在报告结果方面获得了许多奖项和奖励。我们有 最近报道了我们新发现的miR-335- 5 p在抑制破骨细胞分化和骨形成中的作用 再吸收我们进一步发现miR-335- 5 p通过与3 'UTR元件结合发挥其抑制作用 IgSF 3（免疫球蛋白超家族，成员3）。igsf 3在破骨细胞形成过程中的表达增加 分化程度与miR-335- 5 p的表达呈负相关。这一新发现与 miR-335- 5 p的合成代谢成骨作用已得到充分表征，这使我们推测miR-335- 5 p是一种有效的骨诱导剂。 用于治疗骨质疏松症及其相关的骨疾病的药物候选物，其中 骨形成和再吸收受到干扰。目前，骨质疏松症的治疗包括抗吸收药物 和合成代谢骨形成药物然而，这些药物的目标要么是骨吸收，要么是骨形成 路，但不是两者。许多基于蛋白质的疗法具有缺点，包括副作用和高毒性。 成本我们已经产生了miR-335- 5 p基因敲除（功能丧失）和过表达 （功能获得性）小鼠。这两条鼠标线将提供最先进和最复杂的方法 来实现我们的研究目的与科学家和生物工程师合作， 新泽西理工学院，我们已经开发出新颖的和尖端的靶向纳米粒子， 第一次将miR-335- 5 p精确地递送到靶细胞，在靶细胞中它可以发挥其双重作用， 骨吸收和骨形成途径。目标1.探讨新的抗肿瘤药物的分子机制 发现miR-335- 5 p在抑制破骨细胞活性和骨吸收中的功能;目的2.使用我们 新产生的miR-335- 5 p基因敲除和过表达小鼠，以表征多层功能 miR-335- 5 p在骨代谢中的作用;目的3.应用新开发的靶向纳米粒子 miR-335- 5 p对特定细胞类型的作用，并确定其对骨创伤愈合和逆转骨损伤的治疗作用。 骨质疏松我们将确定miR-335- 5 p的治疗效果给定的有效浓度，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生物体中的功能 程度.这个更新项目在概念、技术和干预方面都是创新的。的有利 基于miRNA的治疗的特征将允许这种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骼疾病。的跨学科团队 具有互补和协同技能的研究人员将进行研究（Jake Chen -实验 病理学与骨生物学;齐胜涂-细胞与分子生物学;徐晓阳-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).