title: Genome Editing of Lineage Determinants in Human Pluripotent Stem Cells Reveals Mechanisms of Pancreatic Development and Diabetes.
abstract: Directed differentiation of human pluripotent stem cells (hPSCs) into somatic counterparts is a valuable tool for studying disease. However, examination of developmental mechanisms in hPSCs remains challenging given complex multi-factorial actions at different stages. Here, we used TALEN and CRISPR/Cas-mediated gene editing and hPSC-directed differentiation for a systematic analysis of the roles of eight pancreatic transcription factors (PDX1, RFX6, PTF1A, GLIS3, MNX1, NGN3, HES1, and ARX). Our analysis not only verified conserved gene requirements between mice and humans but also revealed a number of previously unsuspected developmental mechanisms with implications for type 2 diabetes. These include a role of RFX6 in regulating the number of pancreatic progenitors, a haploinsufficient requirement for PDX1 in pancreatic β cell differentiation, and a potentially divergent role of NGN3 in humans and mice. Our findings support use of systematic genome editing in hPSCs as a strategy for understanding mechanisms underlying congenital disorders.
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; 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; Immunology and Microbial Pathogenesis Program, Weill Cornell Graduate School of Medical Sciences, Cornell University, 1300 York Avenue, New York, NY 10065, USA.
authors.forename: Kihyun
authors.initials: K
authors.lastname: Lee
authors.affiliation: Biochemistry, Cell and Molecular Biology Program, Weill Cornell Graduate School of Medical Sciences, Cornell University, 1300 York Avenue, New York, NY 10065, USA.
authors.forename: Bess P
authors.initials: BP
authors.lastname: Rosen
authors.affiliation: Developmental Biology Program, Sloan Kettering Institute, 1275 York Avenue, New York, NY 10065, USA.
authors.forename: Federico
authors.initials: F
authors.lastname: González
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: 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: D001483:Base Sequence; D051792:Basic Helix-Loop-Helix Proteins / Q000378:metabolism; D064113:CRISPR-Cas Systems / Q000235:genetics; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D019070:Cell Lineage / Q000187:drug effects / Q000235:genetics; D003920:Diabetes Mellitus / Q000473:pathology*; D000072669:Gene Editing; D055786:Gene Knockout Techniques; D015894:Genome, Human; D005947:Glucose / Q000494:pharmacology; D057895:Haploinsufficiency / Q000187:drug effects; D018398:Homeodomain Proteins / Q000378:metabolism; D000066449:Human Embryonic Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D007231:Infant, Newborn; D050417:Insulin-Secreting Cells / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D009154:Mutation / Q000235:genetics; D009419:Nerve Tissue Proteins / Q000378:metabolism; D010179:Pancreas / Q000196:embryology* / Q000473:pathology*; D010641:Phenotype; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D000071701:Regulatory Factor X Transcription Factors / Q000378:metabolism; D013997:Time Factors; D015534:Trans-Activators / Q000378:metabolism
chemical_list: D051792:Basic Helix-Loop-Helix Proteins; D005947:Glucose; D018398:Homeodomain Proteins; D009419:Nerve Tissue Proteins; D000071701:Regulatory Factor X Transcription Factors; D015534:Trans-Activators; C498695:NEUROG3 protein, human; C547964:Rfx6 protein, human; C084010:pancreatic and duodenal homeobox 1 protein