Genome editing in hPSCs reveals GATA6 haploinsufficiency and a modifying genetic interaction with GATA4 in human pancreatic development

• Published in: Cell stem cell Vol. 20; no. 5; pp. 675 - 688.e6
• Main Authors: Shi, Zhong-Dong, Lee, Kihyun, Yang, Dapeng, Amin, Sadaf, Verma, Nipun, Li, Qing V., Zhu, Zengrong, Soh, Chew-Li, Kumar, Ritu, Evans, Todd, Chen, Shuibing, Huangfu, Danwei
• Format: Journal Article
• Language: English
• Published: 09.02.2017
• ISSN: 1934-5909; 1875-9777
• DOI: 10.1016/j.stem.2017.01.001

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 .