title: Genome Editing in hPSCs Reveals GATA6 Haploinsufficiency and a Genetic Interaction with GATA4 in Human Pancreatic Development.
abstract: 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 (clustered regularly interspaced short palindromic repeat)/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 it establishes an approach for identifying genetic modifiers of human disease.
authors.affiliation: Developmental Biology Program, Sloan Kettering Institute, 1275 York Avenue, New York, NY 10065, USA.
authors.forename: Zhong-Dong
authors.initials: ZD
authors.lastname: Shi
authors.affiliation: Developmental Biology Program, Sloan Kettering Institute, 1275 York Avenue, New York, NY 10065, USA; Weill Graduate School of Medical Sciences at Cornell University, 1300 York Avenue, New York, NY 10065, USA.
authors.forename: Kihyun
authors.initials: K
authors.lastname: Lee
authors.affiliation: Developmental Biology Program, Sloan Kettering Institute, 1275 York Avenue, New York, NY 10065, USA.
authors.forename: Dapeng
authors.initials: D
authors.lastname: Yang
authors.affiliation: Weill Graduate School of Medical Sciences at Cornell University, 1300 York Avenue, New York, NY 10065, USA.
authors.forename: Sadaf
authors.initials: S
authors.lastname: Amin
authors.affiliation: Developmental Biology Program, Sloan Kettering Institute, 1275 York Avenue, New York, NY 10065, USA; Tri-Institutional MD-PhD Program, Weill Graduate School of Medical Sciences at Cornell University, The Rockefeller University, Sloan Kettering Institute, 1300 York Avenue, New York, NY 10065, USA.
authors.forename: Nipun
authors.initials: N
authors.lastname: Verma
authors.affiliation: Developmental Biology Program, Sloan Kettering Institute, 1275 York Avenue, New York, NY 10065, USA; Louis V. Gerstner Jr. Graduate School of Biomedical Sciences, Memorial Sloan Kettering Cancer Center, 1275 York Avenue, New York, NY 10065, USA.
authors.forename: Qing V
authors.initials: QV
authors.lastname: Li
authors.affiliation: Developmental Biology Program, Sloan Kettering Institute, 1275 York Avenue, New York, NY 10065, USA.
authors.forename: Zengrong
authors.initials: Z
authors.lastname: Zhu
authors.affiliation: Developmental Biology Program, Sloan Kettering Institute, 1275 York Avenue, New York, NY 10065, USA.
authors.forename: Chew-Li
authors.initials: CL
authors.lastname: Soh
authors.affiliation: Department of Surgery, Weill Cornell Medical College, New York, NY 10065, USA.
authors.forename: Ritu
authors.initials: R
authors.lastname: Kumar
authors.affiliation: Department of Surgery, Weill Cornell Medical College, New York, NY 10065, USA.
authors.forename: Todd
authors.initials: T
authors.lastname: Evans
authors.affiliation: Department of Surgery, Weill Cornell Medical College, New York, NY 10065, USA. Electronic address: shc2034@med.cornell.edu.
authors.forename: Shuibing
authors.initials: S
authors.lastname: Chen
authors.affiliation: Developmental Biology Program, Sloan Kettering Institute, 1275 York Avenue, New York, NY 10065, USA. Electronic address: huangfud@mskcc.org.
authors.forename: Danwei
authors.initials: D
authors.lastname: Huangfu
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000235:genetics / Q000502:physiology; D002460:Cell Line; D005455:Fluorescent Antibody Technique; D050991:GATA4 Transcription Factor / Q000235:genetics*; D050993:GATA6 Transcription Factor / Q000235:genetics*; D000072669:Gene Editing / Q000379:methods*; D057895:Haploinsufficiency / Q000235:genetics / Q000502:physiology; D006801:Humans; D008297:Male; D010179:Pancreas / Q000166:cytology / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism*
chemical_list: D050991:GATA4 Transcription Factor; C484170:GATA4 protein, human; D050993:GATA6 Transcription Factor; C494759:GATA6 protein, human
keywords: CRISPR/Cas9 genome editing; GATA6 and GATA4; definitive endoderm; genetic modifier; haploinsufficiency; human embryonic stem cells; human pluripotent stem cells disease modeling; insulin producing pancreatic beta cells; pancreatic agenesis and neonatal diabetes; pancreatic progenitor