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.

Details

Title
Rapid neurogenesis through transcriptional activation in human stem cells
Author
Busskamp, Volker 1 ; Lewis, Nathan E 2 ; Guye, Patrick 3 ; Ng, Alex HM 4 ; Shipman, Seth L 1 ; Byrne, Susan M 1 ; Sanjana, Neville E 5 ; Murn, Jernej 6 ; Li, Yinqing 3 ; Li, Shangzhong 7 ; Stadler, Michael 8 ; Weiss, Ron 3 ; Church, George M 1 

1  Department of Genetics, Harvard Medical School, Boston, MA, USA; Wyss Institute for Biologically Inspired Engineering at Harvard University, Boston, MA, USA 
2  Department of Genetics, Harvard Medical School, Boston, MA, USA; Wyss Institute for Biologically Inspired Engineering at Harvard University, Boston, MA, USA; Department of Biology, Brigham Young University, Provo, UT, USA; Department of Pediatrics, University of California, San Diego, CA, USA 
3  Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA 
4  Department of Genetics, Harvard Medical School, Boston, MA, USA; Wyss Institute for Biologically Inspired Engineering at Harvard University, Boston, MA, USA; Department of Systems Biology, Harvard Medical School, Boston, MA, USA 
5  Broad Institute of MIT and Harvard, Cambridge Center, Cambridge, MA, USA; McGovern Institute for Brain Research, Department of Brain and Cognitive Sciences, Department of Biological Engineering, Massachusetts Institute of Technology (MIT), Cambridge, MA, USA 
6  Department of Cell Biology, Harvard Medical School, Boston, MA, USA; Division of Newborn Medicine, Boston Children's Hospital, Boston, MA, USA 
7  Department of Bioengineering, University of California, San Diego, CA, USA 
8  Friedrich Miescher Institute for Biomedical Research, Basel, Switzerland; Swiss Institute of Bioinformatics, Basel, Switzerland; University of Basel, Basel, Switzerland 
Section
Articles
Publication year
2014
Publication date
Nov 2014
Publisher
EMBO Press
e-ISSN
17444292
Source type
Scholarly Journal
Language of publication
English
DOI
https://doi.org/10.15252/msb.20145508
ProQuest document ID
2290149842
Copyright
© 2014. This work is published under http://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.