lnc-A0介导DC-STAMP/NFATc1负反馈环路调控破骨细胞前体在TEB修复中的作用机制研究

阐释组织工程骨（TEB）修复中的骨重建机制，是探索TEB临床应用的重要命题。破骨细胞（OC）作为骨重建的要素之一，其参与该过程的机理不详，尤其是OC前体融合的调控研究甚少，故以此切入探求TEB骨重塑机制极为必要。在证实关键融合分子DC-STAMP可下调其上游转录分子NFATc1的基础上，基于DC-STAMP为变量的lncRNA芯片结果提示lnc-AK077216 (lnc-A0)可靶向调控NFATc1,因此提出“lnc-A0介导DC-STAMP/NFATc1负反馈环路调控OC前体融合”假说。拟通过荧光素酶基因报告系统、RIP-seq等证实前述靶向关系；利用ChIRP-seq等技术证实该环路，并观察其对OC前体分化及融合的效应。利用小鼠TEB修复模型，在体观察lnc-A0对环路分子的调控,检测干预lnc-A0表达对骨重建的影响。本研究有利于阐明OC分化机理，为TEB的构建和修复拓展新思路

To better elucidate the detailed mechanism of bone remodeling in Tissue Engineering Bone (TEB) is considered to be one of the main issues before large-scale clinical application. Osteoclasts (OCs) are crucial players during bone remodeling, however, the mechanism of osteoclastic function in TEB-mediated bone repair remains unclear. Particularly, the role of cell-cell fusion among osteoclast precursors (OCPs) in bone remodeling is rarely studied. Therefore, our group focused on the role of OCPs fusion as part of the mechanism in TEB-mediated bone remodeling. We previously confirmed that the key fusogenic protein DC-STAMP could down regulate its upstream molecule NFATc1 in turn. Recently, our LNCRNA microarray results indicated that lnc-AK077216 exspression was directly associated with NFATc1. Based on the evidence above, we hypothesized that lnc-AK077216 is involved in the negative feedback loop of “DC-STAMP/NFATc1” potentially playing a key role in osteoclastogenesis. To verify our hypothesis, we intend to verify this targeting relationship by dual luciferase gene report system and RIP-seq. Techniques such as ChIRP-seq will be adopted to certify the hypothesized feedback loop and to observe OCPs proliferation, differentiation and especially fusion process. Furthermore, lnc-AK077216 mediated loop will be tested during bone repairing by TEB in mice with bone defect. We deem our study will further illustrate the mechanism of cell-cell fusion of osteoclasts and their role in TEB-mediated bone remodeling, and thus provide novel strategies and methods in building more efficient TEB.