PARP活性对人胚胎干细胞向视网膜色素上皮细胞定向分化的作用

项目名称： PARP活性对人胚胎干细胞向视网膜色素上皮细胞定向分化的作用

项目编号： No.31201108

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 发育生物学与生殖生物学

项目作者： 高芙蓉

作者单位： 同济大学

项目金额： 21万元

中文摘要： 人胚胎干细胞（hESCs）可为视网膜色素上皮细胞（RPE）移植治疗视网膜变性疾病提供无限细胞来源。然而，有效诱导hESCs向RPE分化的因子仍然不清楚。我们前期研究发现PARP活性抑制剂PJ34促进hESCs向神经前体细胞分化，进而促进RPE分化；抑制PARP活性显著增加Sox2蛋白水平。文献报道Sox2和Pax6都是神经前体细胞的决定因子，且PARP调控Pax6的转录，抑制PARP活性能保护神经前体细胞的存活。综上，我们提出假设：抑制PARP活性通过调节Sox2或Pax6并抑制凋亡，最终促进RPE分化。本研究拟在前期工作基础上，采用自主建立的hES细胞系SHhES2，利用PJ34诱导hESCs向RPE分化，通过基因过表达或RNA干扰等技术以及基因芯片和凋亡检测研究PJ34促进RPE分化的机制，为诱导hESCs向RPE分化提供指导和新的途径，并对揭示视网膜发育和视网膜变性的机制有重要意义。

中文关键词： 人胚胎干细胞；视网膜色素上皮细胞；分化；PARP活性；PAX6

英文摘要： Human embryonic stem cells (hESCs) may serve as an unlimited source of retinal pigment epithelium (RPE) cells for transplantation in retinal degeneration diseases. However, the differentiation process of hESCs to RPE is not well defined and most often necessitates a long-term differentiation time with low efficiency; the ideal supplements for the induction of directed differentiation are still unclear. During our previous work, we found that addition of PJ34, an inhibitor of PARP activity, promotes the differentiation of hESCs to neuroectodermal progenitors and RPE, and increases Sox2 protein level.In addition, it is reported that Sox2 and Pax6 are required for neuroectodermal progenitors, PARP regulates transcription of Pax6, and PARP inhibition rescues hESC-derived neural progenitors from cell death.In view of the above,we hypothesized that PJ34 regulates Sox2 and Pax6 and inhibits cell apoptosis to promote differentiation of RPE from hESCs.In this proposal, we planed to use SHhES2, a hES cell line established by ourselves, to differentiate into RPE cells by PJ34. Then, we will study the role of PJ34 in differentiation through activation or attenuation of function of Sox2 and Pax6 and examination of apoptosis during RPE differentiation in the presense or absence of PJ34. Our study will not only supply a new wa

英文关键词： human embryonic stem cells；retinal pigment epithelium；differentiation；PARP acitivty；PAX6