多发性骨髓瘤微泡横向传递分泌型"miRNAs分子群"抑制BMSC成骨分化

Multiple myeloma(MM) cells continuously reprogramme bone marrow mesenchymal stem cells(BMSC), which plays a key role in pregression and incurable properties of MM. With the support of the previous National Science Grant(No.81071943), we have found that myeloma cell-derived microvesicles(MM-MV) can seriously inhibit the osteoblast differentiation of BMSC and act as a novel communication pathway of myeloma cells to BMSCs. Since microvesicle enclosed secreted miRNAs owns the advantages of both "signal cluster" and "targeting", we propose that the secreted "miRNA signal cluster" in MM-MV plays a key role in reprogramming BMSC by horizonal transfer of these miRNAs into BMSC. In this study, firstly, the characters of MM-MV enclosed miRNAs and its effects as a whole on the osteoblast defferentiation of BMSC will be investigated. Secondly, the categories and quantity of these miRNAs will be confirmed by deep sequencing and differentiation analysis of miRNA expressions. Then, with analysis of the miRNA- TF feed-forward loops, the key nodal miRNAs will be picked out in the complicated network. Finally, both the effects of these key miRNAs on the osteoblast differentiation of BMSC and the potential of being clinic specific markers will be evaluated by in vitro and in vivo experiments and clinical investigations, respectively. This work is necessary to the our original findings of the previous grant. It will help to clarificate the main molecuar mechanisms with the advantage of interdisciplinary research. This study will lead to finding novel mechanisms in tumorgenesis and progression of myeloma, is to possibly provide novel targets for treatment and specific markers to myeloma.

多发性骨髓瘤（MM）细胞改造BMSC是MM微环境维持其进展和不可治愈的关键事件。前期项目（No.81071943）证实MM来源微泡（MM-MV）显著抑制BMSC成骨分化，是MM胞间新型通讯途径和信息载体，基于近期MV分泌型miRNA研究的重要发现，我们推测MM-MV内分泌型"miRNAs分子群"可横向传递进入BMSC，进而以交互性、多靶向网络方式抑制其成骨分化。本项目拟首先研究MM-MV分泌型"miRNAs分子群"富集模式，及其横向传递后对BMSC成骨分化的综合影响；进而通过深度测序锁定传入的miRNAs，并采用前期成功运用的"miRNA-TF共调控FFL"预测方法，从中遴选出关键调控作用的节点miRNAs；最后通过体外、体内功能实验予以验证，并探讨其临床标志物意义。本项目依据学科交叉，深入研究前期研究结果的分子机制，旨在揭示MM异质性进展规律，为临床提供特异性标志物和新型干预靶标。

多发性骨髓瘤（MM）细胞改造BMSC是MM微环境维持其进展和不可治愈的关键事件，为了研究MM细胞来源微泡（MM-MVs）在其中的作用，我们在前期项目（No.81071943）中证实了体外和动物体内证实MM-MVs可显著抑制BMSC成骨分化。在此基础上，本项目证实了MM-MVs促进EC血管新生，显示了MM细胞通过MM-MVs促进肿瘤血管新生的新途径和机制；同时，我们研究还发现在应激状态下，MM细胞分泌更多特异性MM-MVs促进MM细胞躲避应激打击，促进应激态下的肿瘤血管新生和肿瘤细胞存活。我们在该项目资助下，关注并研究了MM-MVs内分泌型"miRNAs分子群"的富集规律及其可能作用靶标，摸索并建立了临床MM患者外周血和尿液中MM-MVs的提取和流式检测的程序方法，重点研究了MM-MVs与临床疾病进展、骨病、肾病等特征性标志间的相关性和临床意义。本项目依据学科交叉， 深入研究MM-MVs的相关作用和分子机制，证实了MM-MV在MM的肿瘤及骨病（MBD）中的标志物意义，为临床提供潜在特异性标志物和新型干预靶标。本项目全部或部分支持培养研究生8人，其中博士生3人，硕士生5人，共计发表论文8篇，其中SCI收录7篇，获省部级一等奖一项。