Genome Editing in hPSCs Reveals GATA6 Haploinsufficiency and a Genetic Interaction with GATA4 in Human Pancreatic Development.
Genome Editing in hPSCs Reveals GATA6 Haploinsufficiency and a Genetic Interaction with GATA4 in Human Pancreatic Development.
复制标题
DOI：
10.1016/j.stem.2017.01.001
复制
发表时间：
2017-05-04
期刊：
影响因子：
23.9
通讯作者：
Huangfu D
中科院分区：
文献类型：
--
作者：
Shi ZD;Lee K;Yang D;Amin S;Verma N;Li QV;Zhu Z;Soh CL;Kumar R;Evans T;Chen S;Huangfu D
与单倍体基因需求不足相关的人类疾病表型通常无法在动物模型中得到很好的重现。在这里，我们使用 CRISPR/Cas9 介导的基因组编辑结合人类多能干细胞 (hPSC) 定向分化，研究了人类 GATA6 单倍体不足与多种临床表型之间的关联，其中包括新生儿和成人发病的糖尿病。我们发现，一个 GATA6 等位基因的缺失会特别影响人类胰腺祖细胞从早期 PDX1+ 阶段向更成熟的 PDX1+NKX6.1+ 阶段的分化，导致葡萄糖反应性 β 样细胞的形成受损。除了GATA6单倍体不足之外，我们还确定了GATA6和GATA4在定形内胚层和胰腺祖细胞形成中的剂量敏感要求。我们的工作将 hPSC 的应用范围从研究单个基因位点的影响扩展到研究多基因人类性状，并建立了一种识别人类疾病遗传修饰因子的方法。 Huangfu、Chen 及其同事通过逐步分化转基因 hPSC 来模拟人类胰腺疾病，以表征在小鼠模型中不明显的 GATA6 单倍体不足的表型效应，以及与 GATA4 的遗传相互作用。
Human disease phenotypes associated with haploinsufficient gene requirements are often not recapitulated well in animal models. Here, we have investigated the association between human GATA6 haploinsufficiency and a wide-range of clinical phenotypes that include neonatal and adult-onset diabetes using CRISPR/Cas9-mediated genome editing coupled with human pluripotent stem cell (hPSC) directed differentiation. We found that loss of one GATA6 allele specifically affects the differentiation of human pancreatic progenitors from the early PDX1+ stage to the more mature PDX1+NKX6.1+ stage, leading to impaired formation of glucose-responsive β-like cells. In addition to this GATA6 haploinsufficiency, we also identified dosage-sensitive requirements for GATA6 and GATA4 in the formation of both definitive endoderm and pancreatic progenitor cells. Our work expands the application of hPSCs from studying the impact of individual gene loci to investigation of multigenic human traits, and establishes an approach for identifying genetic modifiers of human disease. Huangfu, Chen and colleagues model human pancreatic disease by step-wise differentiation of genetically modified hPSCs to characterize phenotypic effects of GATA6 haploinsufficiency not evident in mouse models plus genetic interaction with GATA4.