LncRNAs-miRNAs交互作用调控人脂肪干细胞成骨分化及其机制的研究

Maxillofacial bone defects severely impact patients’physical and mental health, and the common therapeutic method of bone defects repair is autogenous bone graft which has limitations. Therefore, bone tissue engineering has become a hot topic and one of the key points is to induce osteogenic differentiation of seed cells safely and efficiently. Most studies employed releasing osteogenic factors in bone defects or conducting gene transduction, but there are some limitations. Recent studies found that specific long noncoding RNAs (lncRNAs) regulated osteogenic differentiation of mesenchymal stem cells (MSCs), and lncRNAs normally bind with microRNA (miRNAs) to function, which might be a breakthrough of regulation of osteogenic differentiation. Our previous studies indicated that expression of several lncRNAs can be increased or decreased significantly in the osteogenic differentiation process of human adipose derived-stem cells (hASCs). However, the interaction among lncRNAs and miRNAs in regulation of osteogenic differentiation of hASCs remains unknown. Based on our previous studies, this project will use microarray technology, series experiments in vitro and in vivo, and methods of mechanism study to confirm the key lncRNAs that regulate the osteogenic differentiation of hASCs, identify the miRNAs that interact with lncRNAs, investigate the effects and mechanisms of the interacted lncRNAs-miRNAs regulation of hASC osteogenic differentiation, and explore the potential molecular targets associated with several key osteogenic signaling pathways. This project is expected to provide a novel way for maxillofacial bone defects repair.

颌面部骨缺损严重影响患者身心健康，常用自体骨移植修复但有其局限，采用骨组织工程方法为现研究热点，但如何安全高效定向诱导种子细胞成骨分化是一核心问题，多采用在骨缺损局部释放促成骨相关因子或通过基因转染等方法但存在一些问题。最新文献报道lncRNAs可以明显地调控间充质干细胞的成骨分化，并通常与miRNAs交互协调发挥作用，有望成为新的突破点。课题组前期研究也证实人脂肪干细胞成骨分化过程相关lncRNAs表达明显上调或下调，但尚未见其交互作用调控人脂肪干细胞成骨分化的报道。本项目拟通过基因芯片筛选、系列实验确认、体外及体内验证、深入探索机制的技术路线确定在人脂肪干细胞成骨分化中调控作用最佳的lncRNAs，研究并鉴定与之交互作用的miRNAs，明确配对的lncRNAs-miRNAs调控脂肪干细胞成骨分化的机制，并研究该交互作用与成骨分化的重要信号通路产生作用的分子靶点，为骨缺损修复提供新思路。

颌面部骨缺损严重影响患者身心健康，常用的自体骨移植修复发放有一定局限性，采用骨组织工程方法为目前研究热点，而如何促进干细胞定向分化并维持其表型稳定是再生医学研究的核心问题。已有证据表明非编码RNA如lncRNAs、circRNAs、miRNAs等在干细胞成骨分化中扮演重要角色。课题组前期发现在人脂肪间充质干细胞成骨分化过程中有众多lncRNAs出现差异表达，但其具体调控机制尚不清楚。本项目探究人脂肪干细胞成骨分化相关的关键lncRNA及与其有密切交互作用的miRNA，并研究lncRNA-miRNA网络的交互作用机制及下游重要信号通路，有望利用该机制调控脂肪干细胞的成骨分化效果。本项目的执行阐明了人脂肪间充质干细胞成骨分化过程中，关键lncRNA DANCR、lncRNA H19、lncRNA NIOAT通过结合相应miRNAs发挥调控作用的机制，并初步探索了circPOMT1和circMCM3AP在人脂肪间充质干细胞成骨分化中的作用。同时，本项目初步探索发现的circINO80在人牙髓干细胞成骨分化中具有应力敏感特性。本项目为靶向调控非编码RNA以促进骨缺损修复提供了理论基础及新思路，并为后续探究应力敏感circINO80调控人牙髓干细胞成骨分化修复颌骨缺损奠定了工作基础。