基于GATA2敲除及GATA2-GFP标记的hES细胞系研究内皮-造血转变的调控机制

项目名称： 基于GATA2敲除及GATA2-GFP标记的hES细胞系研究内皮-造血转变的调控机制

项目编号： No.31500948

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 生物科学

项目作者： 杜鹃

作者单位： 中国科学院广州生物医药与健康研究院

项目金额： 20万元

中文摘要： 明确人早期造血发育的调控机制，对通过体外定向分化途径获得人的功能性造血干细胞(HSC)至关重要。早期造血发育中，内皮-造血转变(EHT)过程是重要节点之一。基于小鼠模型研究发现，GATA2在EHT过程中起重要的调控作用，但迄今未有GATA2在人的造血发育EHT的平行研究报导。鉴于人鼠造血发育的差异性，本课题组直接选择以人的GATA2敲除和内源GATA2-GFP标记的ES细胞系体外造血分化模型来研究EHT调控。本课题拟通过比较GATA2敲除、GFP标记敲入情况下，体外造血分化获得的生血内皮、造血干祖细胞的转录组，寻找GATA2在EHT调控过程中的效应因子和共同作用因子，并通过功能补偿和shRNA敲除等手段加以验证，来揭示GATA2相关的EHT调控机制，完善对人类早期造血发育分化调控的认识，从而为多能干细胞定向造血分化获得HSC奠定基础。

中文关键词： GATA2；内皮-造血转变；生血内皮；分子机制

英文摘要： The inability of obtaining hematopoietic stem cell (HSC) capable of long-term reconstitution and full lineage specification is partially due to lack of understanding about hematopoiesis during early embryo development. Among the early events of early hematopoiesis, endothelial-to-hematopoietic-transition (EHT) is a key time point. Data from Gata2 knock-out mouse model suggest that GATA2 plays an important regulatory role during EHT. However, parallel study in human hematopoiesis is still missing. Due to the difference between human and mouse hematopoiesis, we chose GATA2 as point of entry to study mechanism of EHT in human system. We have recently established both GATA2 knock-out and GATA2-GFP knock-in hES cell line and plan to compare transcriptome of HE and HSPC obtained from in vitro hematopoietic differentiation of these cell lines. We aim to reveal the regulatory mechanism of EHT by discovering GATA2 effectors and transcription factors cooperating with GATA2. Functional tests such as function rescue or shRNA knockdown were also planned to validate the regulatory role of these effectors and transcription factors. Dissection of mechanism of EHT regulation will pave our way to fully understand hematopoiesis regulation during early human embryo development and eventually lead to high efficient method for generation of transplantable HSC through in vitro differentiation of pluripotent stem cells.

英文关键词： GATA2;EHT;HE;Molecular mechanism