抗体诱导成纤维细胞向肝实质细胞直接重编程的靶点及机制研究

项目名称： 抗体诱导成纤维细胞向肝实质细胞直接重编程的靶点及机制研究

项目编号： No.31501195

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 发育生物学与生殖生物学

项目作者： 王菊

作者单位： 上海科技大学

项目金额： 20万元

中文摘要： 高等动物的成体细胞之间能不通过多能性干细胞阶段而被直接重编程。通过过表达肝脏特异的转录因子组合，能够诱导成纤维细胞重编程成为类肝实质细（iHep）。iHep 细胞具有与肝实质细胞类似的基因表达谱与部分肝细胞功能，重编程得到的肝实质细胞可为肝脏发育和肝脏疾病的研究提供新的模型。然而，成体细胞的重编程都需要转录因子的协同作用，病毒的应用极大的限制了此类方法的发展前景。另外，现有的重编程方法获得的肝实质细胞在体外扩增能力较弱，肝细胞功能不全。而且关于重编程的靶点与作用机制还没有清楚的阐述。因此，我们拟建立抗体诱导的成纤维细胞向肝实质细胞的直接重编程技术。通过细胞内组合抗体库技术，以及高通量的流式细胞分选方法，筛选获得能够诱导成纤维细胞向肝实质细胞直接重编程的激动型抗体，并通过受体抗体的特异性结合，对重编程相关靶点与作用机制进行深入探讨。

中文关键词： 小鼠；重编程；肝实质细胞；类肝实质细胞；抗体

英文摘要： Hepatocytes are the most abundant hepatic cells playing a significant role in the functions of liver. The generation of large numbers of functional hepatocytes for cell-based approaches to liver disease and scientific research is an important and unmet goal. Terminally differentiated somatic cells can be directly reprogrammed into other lineages without reversion to a stem cell state, by overexpressing lineage-specific transcription factors. Direct reprogramming of fibroblasts to hepatic lineages could offer functional hepatocytes to this problem without immunologic rejection and potential tumorigenicity. However all of them achieved by transduction of combination of definite transcriptional factors, in which virus not permitted for clinical treatment were employed. Besides, despite the great potential of these new technologies to obtain hepatocyte-like cells, little progresses have been made to push their applications forward to clinics or industries because of the limited function and expansion in vitro. Moreover, the target and mechanism is still not demonstrated clearly. Antibody is a novel molecular with great promise. We design to establish new methods for direct programming of fibroblast to hepatocyte-like cells by antibodies selected from the intracellular combinatorial antibody library approach. It has been reported that agonist antibody induced receptor pleiotropia for direct programming of cell fate. The agonist antibody not present in nature may give a different dimension to the important problem of signal transduction. Exploring the target and mechanism of the direct cell fate conversion with antibodies is worth looking forward to.

英文关键词： mouse;direct reprogramming;hepatocyte;iHep cell;antibody