DLX3基因修饰iPS细胞膜片在放疗后种植体骨愈合中的作用及机制研究

DLX3 gene plays critical role in regulating directional osteogenic differentiation of stem cells. iPS cells possess the capability of multi-differentiation, particularly osteogenic differentiation, but the mechanism was not clear. Our group has established animal model for osseointegration in irradiated bone, extracted renal epithelial cells from human urine, reprogrammed those into iPS cells and formed iPS cell sheet. This project is proposed to reprogramme renal epithelial cells into iPS cells, construct DLX3 modified iPS cells (iPS/DLX3 cells), analyze proliferation and differentiation of iPS/DLX3 cells, predict and testify downstream target genes of DLX3. Additionally, this project is to prepare iPS/DLX3 cell sheet, apply the cell sheet onto dental implant surface, investigate osseointegration of iPS/DLX3 cell sheet compounded implant in irradiated site and investigate the mechanism of the changes. This study will be of great value in exploring target and downstream signal pathway of osseointegration in irradiated site, and be guidance significant for iPS directional osteogenic differentiation.

DLX3基因在干细胞向成骨细胞分化中起重要的调控作用。诱导性多能干细胞具备多向分化潜能，可向成骨细胞分化，但其机制尚不清楚。本课题组前期已建立放疗动物种植体骨愈合模型、从尿液提取肾上皮细胞并重编程为iPS细胞、成功形成iPS细胞膜片。本项目拟以人尿液提取肾上皮细胞，重编程为iPS细胞，进而构建DLX3基因重组iPS细胞(iPS/DLX3细胞)。分析iPS/DLX3细胞的增殖及分化情况，通过RNA-seq及生物信息学技术验证DLX3下游靶基因。构建DLX3基因修饰的iPS细胞膜片，并将其用于放疗区域的种植体表面，分析探讨DLX3基因在放疗情况下促进骨整合的作用及机制。本研究旨在探索促进放疗后骨整合的靶点及下游信号，并为调控iPS细胞向成骨细胞分化的调控提供理论依据。

DLX3基因在干细胞向成骨细胞分化中起重要的调控作用。诱导性多能干细胞具备多向分化潜能，可向成骨细胞分化，但其机制尚不清楚。本课题组前期已建立放疗动物种植体骨愈合模型、从尿液提取肾上皮细胞并重编程为iPS细胞、成功形成iPS细胞膜片。本项目拟以人尿液提取肾上皮细胞，重编程为iPS细胞，进而构建DLX3基因重组iPS细胞(iPS/DLX3细胞)。分析iPS/DLX3细胞的增殖及分化情况，通过RNA-seq及生物信息学技术验证DLX3下游靶基因。构建DLX3基因修饰的iPS细胞膜片，并将其用于放疗区域的种植体表面，分析探讨DLX3基因在放疗情况下促进骨整合的作用及机制。本课题组研究发现：（1）DLX3基因转染的iPS细胞来源的间充质干细胞具有更强的成骨能力，而增殖能力减弱。（2）RNA-seq发现DLX3可通过激活DSP、MEPE、OCN、Runx2促进iPS细胞来源间充质干细胞成骨向分化。