iPSC联合TALEN基因组编辑技术探索子痫前期的发病机制及药物筛选

Perfect combination of induced pluripotent cells (iPSCs), genetic editing and whole genome technology give us opportunity to model disease，genetic modification and explore the pathogenesis. There are many successful disease modelling using the above technology. We studied the early pathology using iPSCs from ALS patients, on basis of the experience in above study, this project plans to apply iPSCs differentiation into trophoblast cells, and dynamically explore the early pathology and the abnormal pathway of eclampsia, which greatly threaten the health of mother and infant, and finally do new drug screen using high throughput technology. This study supply a novel strategy and therapy for those disease with abnormal development of trophoblastic cells including pre-eclampsia, fetal growth restriction, recurrent spontaneous abortion and repeated implantation failure!

多潜能干细胞（iPSC），基因组编辑，全基因组技术的完美结合，为我们提供了在相关人类细胞中模拟疾病，遗传改造及探索发病机制的难得机遇。相关技术已在许多疾病研究中获得了成功。我们利用上述技术成功探索了脊髓侧索硬化症的早期病理改变。在此基础上本项目将利用iPSC向滋养细胞分化这一成熟且简便的分化体系，从源头上从分子及细胞水平连续动态解析子痫前期这一严重危害母胎健康疾病中滋养细胞的早期病理损害及异常信号通路并进行高通量的新药筛选。本项目的完成将为子痫前期、胎儿生长受限、复发性流产，反复着床失败等滋养细胞发育异常相关疾病提供新的研究策略及治疗思路。

在本项国家自然科学基金资助下,我们获得子痫前期患者特异性的iPSC细胞系2株，正常对照iPSC细胞系1株。鉴定其干细胞特异性的全能性标志物：OCT4、SOX2、NANOG、TRA-1-60均为阳性，并能在体外分化为畸胎瘤。无饲养层条件下加入BMP4分化4天后的细胞表达滋养干细胞标志物CDX2及GATA3，不表达胚胎干细胞标志物OCT4，绒毛外滋养细胞（EVT）标志物HLA-G及合体滋养细胞（STB）标志物HCG。初步判断其为滋养干细胞样细胞。将细胞传代至colIV上，用CTB维持培养基（包含6种重要的小分子EGF，CHIR99021，A83-01，SB431542，VPA ，Y27632）培养，观察细胞可增殖十代左右，获得的细胞滋养干细胞标志物CDH1的阳性比例为88.08%，滋养细胞标志物KRT7的阳性比例为91.38%。本项目的完成将为子痫前期，胎儿生长受限，复发性流产，反复着床失败等滋养细胞发育异常相关疾病提供新的研究策略及治疗思路。