Single-Cell Gene Expression Analysis of a Human ESC Model of Pancreatic Endocrine Development Reveals Different Paths to β-Cell Differentiation

• Published in: Stem cell reports 2017-10, Vol.9 (4), p.1246-1261
• Main Authors: Petersen, Maja Borup Kjær, Azad, Ajuna, Ingvorsen, Camilla, Hess, Katja, Hansson, Mattias, Grapin-Botton, Anne, Honoré, Christian
• Format: Article
• Language: English
• Subjects: Basic Helix-Loop-Helix Transcription Factors - genetics; Basic Helix-Loop-Helix Transcription Factors - metabolism; Biomarkers; Cell Differentiation - genetics; Cell Lineage - genetics; Computational Biology - methods; differentiation; Embryonic Stem Cells - cytology; Embryonic Stem Cells - metabolism; endocrine; Gene Expression Profiling; Gene Expression Regulation, Developmental; Genes, Reporter; Homeodomain Proteins - genetics; Homeodomain Proteins - metabolism; hormone; Humans; Immunophenotyping; Insulin-Secreting Cells - cytology; Insulin-Secreting Cells - metabolism; lineage; Models, Biological; Nerve Tissue Proteins - genetics; Nerve Tissue Proteins - metabolism; NEUROG3; Organogenesis - genetics; pancreas; Phenotype; pluripotent stem cells; progenitor; Single-Cell Analysis; Transcriptome
• ISSN: 2213-6711; 2213-6711
• DOI: 10.1016/j.stemcr.2017.08.009

The production of insulin-producing β cells from human embryonic stem cells (hESCs) in vitro represents a promising strategy for a cell-based therapy for type 1 diabetes mellitus. To explore the cellular heterogeneity and temporal progression of endocrine progenitors and their progeny, we performed single-cell qPCR on more than 500 cells across several stages of in vitro differentiation of hESCs and compared them with human islets. We reveal distinct subpopulations along the endocrine differentiation path and an early lineage bifurcation toward either polyhormonal cells or β-like cells. We uncover several similarities and differences with mouse development and reveal that cells can take multiple paths to the same differentiation state, a principle that could be relevant to other systems. Notably, activation of the key β-cell transcription factor NKX6.1 can be initiated before or after endocrine commitment. The single-cell temporal resolution we provide can be used to improve the production of functional β cells. [Display omitted] •Single-cell qPCR identifies subpopulations on hESC to endocrine differentiation paths•All hESC-derived endocrine cells transcribe multiple hormones in vitro•A subpopulation of hESC-derived INS+ cells transcriptionally resembles adult β cells•NKX6.1 onset before or after endocrine commitment leads to β-cell differentiation In this article, Honoré, Grapin-Botton, and colleagues use single-cell expression profiling to show a differentiation sequence from hESCs to pancreatic endocrine cells and early divergence of paths to different endocrine subtypes. Two paths lead to β-cell differentiation where NKX6.1 can be initiated before or after endocrine commitment.