Rapid neurogenesis through transcriptional activation in human stem cells

Molecular Systems Biology

Volker Busskamp¹  Nathan E Lewis¹  Patrick Guye⁴  Alex HM Ng¹  Seth L Shipman¹  Susan M Byrne¹  Neville E Sanjana²  Jernej Murn⁵  Yinqing Li⁴  Shangzhong Li³  Michael Stadler⁶  Ron Weiss⁴ 

[1] Department of Genetics, Harvard Medical School, Boston, MA, USA;Broad Institute of MIT and Harvard, Cambridge Center, Cambridge, MA, USA;Department of Bioengineering, University of California, San Diego, CA, USA;Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA;Department of Cell Biology, Harvard Medical School, Boston, MA, USA;Friedrich Miescher Institute for Biomedical Research, Basel, Switzerland

关键词: gene regulatory networks;    microRNAs;    neurogenesis;    stem cell differentiation;    transcriptomics;   

DOI  :  10.15252/msb.20145508

来源: Wiley

【 摘 要 】

Abstract

Advances in cellular reprogramming and stem cell differentiation now enable ex vivo studies of human neuronal differentiation. However, it remains challenging to elucidate the underlying regulatory programs because differentiation protocols are laborious and often result in low neuron yields. Here, we overexpressed two Neurogenin transcription factors in human-induced pluripotent stem cells and obtained neurons with bipolar morphology in 4 days, at greater than 90% purity. The high purity enabled mRNA and microRNA expression profiling during neurogenesis, thus revealing the genetic programs involved in the rapid transition from stem cell to neuron. The resulting cells exhibited transcriptional, morphological and functional signatures of differentiated neurons, with greatest transcriptional similarity to prenatal human brain samples. Our analysis revealed a network of key transcription factors and microRNAs that promoted loss of pluripotency and rapid neurogenesis via progenitor states. Perturbations of key transcription factors affected homogeneity and phenotypic properties of the resulting neurons, suggesting that a systems-level view of the molecular biology of differentiation may guide subsequent manipulation of human stem cells to rapidly obtain diverse neuronal types.

Synopsis

Rapid and homogeneous neuronal differentiation is attained in human stem cells upon overexpression of two Neurogenin transcription factors. mRNA and miRNA expression profiling during differentiation reveals a regulatory network mediating neurogenesis from stem cells.

• Neurogenin-1 and -2 drive homogeneous differentiation of human stem cells into bipolar neurons in 4 days in defined media.
• The population homogeneity allowed mRNA and miRNA expression profiling over time during neurogenesis.
• A network of key transcription factors and miRNAs that promote rapid neurogenesis and loss of pluripotency is identified.
• Perturbations of key transcription factors affect the homogeneity and phenotypic properties of the resulting neurons.

【 授权许可】

CC BY   
© 2014 The Authors. Published under the terms of the CC BY 4.0 license

Creative Commons Attribution 4.0 License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.