候选lncRNAs调控脂肪干细胞成髓核分化的作用及机制研究

The reduction of nucleus pulposus cells is usually observed in degenerative disc, which is considered to be the original reason of intervertebral disc degeneration. Based on our group and colleagues’ research work, the harsh microenvironment obviously constrained the transdifferentiation of stem cells into nucleus pulposus cells, which is the bottleneck of the biotherapy treatment for disc degenerative disease. Long noncoding RNA (lncRNA) played a crucial regulatory role in oriented differentiated from stem cells, however, the definite function of lncRNA has not been elucidated. From our lncRNA-seq sequencing data, we found that lncRNA-XLOC_001098, lncRNA-XLOC_000150, lncRNA-XLOC_011378 and lncRNA-XLOC_001078 were key regulatory 1ncRNAs in transdifferentiation of human dipose stem cells (hADSCs) into nucleus pulpous cells, and these 1ncRNAs was closely related with the gene expression of sonic hedgehog signaling pathway (including protein kinase A, zinc finger protein et.al), which is the critical differentiation pathway in differentiation of stem cell. Therefore, the project intends to determine the temporal and spatial distribution characteristics and regulatory mechanisms of difference lncRNA in transdifferentiation of hADSC into nucleus pulposus cells in vitro, and acquires high transdifferentiation 1ncRNA-hADSC cell line into nucleus pulposus cells using gene transfection technology, and then transplants 1ncRNA-hADSC into degenerative rat intervertebral disc in allografts and xenografts method after differentiation. The specific molecular mechanisms of 1ncRNA will be clarifed in transdifferentiation of hADSC into nucleus pulposus cells, which provides experimental data and new targets for stem cell transplantation in the treatment of disc degeneration.

髓核细胞减少是椎间盘退变的始动因素，而椎间盘恶劣微环境下干细胞成髓核分化受限是治疗椎间盘退变的瓶颈。研究发现lncRNA是干细胞定向分化的重要调控因素，但功能尚未阐明。本课题组利用lncRNA-seq测序发现，lncRNA-XLOC_001098、XLOC_000150、XLOC_011378和XLOC_001078可能是人脂肪干细胞(hADSC)成髓核分化的关键1ncRNAs，且与干细胞定向分化的关键通路—音猬因子信号通路多种基因表达紧密相关。基于此，本项目拟在体外确定hADSC成髓核分化中差异lncRNA的时空分布特性和调控机制，基因转染获得高成髓核分化的1ncRNA-hADSC细胞系，进而异种异体移植成髓核分化后的1ncRNA-hADSC到大鼠退变椎间盘内，阐明1ncRNA在hADSC成髓核分化过程中的作用及具体分子调控机制，从而为椎间盘退变的干细胞移植治疗提供实验数据及新靶点。

髓核细胞减少是椎间盘退变的始动因素，而椎间盘恶劣微环境下干细胞成髓核分化受限是治疗椎间盘退变的瓶颈。研究发现lncRNA是干细胞定向分化的重要调控因素，但功能尚未阐明。本课题组利用lncRNA-seq测序发现，lncRNA-XLOC_001098、XLOC_000150、XLOC_011378和XLOC_001078可能是人脂肪干细胞(hADSC)成髓核分化的关键1ncRNAs，且与干细胞定向分化的关键通路—音猬因子信号通路多种基因表达紧密相关。本项目发现阐明1ncRNA在hADSC成髓核分化过程中的作用及具体分子调控机制，从而为椎间盘退变的干细胞移植治疗提供实验数据及新靶点。