Rapid neurogenesis through transcriptional activation in human stem cells.
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.
Department of Genetics, Harvard Medical School, Boston, MA, USA Wyss Institute for Biologically Inspired Engineering at Harvard University, Boston, MA, USA.
Volker
V
Busskamp
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.
Nathan E
NE
Lewis
Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA.
Patrick
P
Guye
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.
Alex H M
AH
Ng
Department of Genetics, Harvard Medical School, Boston, MA, USA Wyss Institute for Biologically Inspired Engineering at Harvard University, Boston, MA, USA.
Seth L
SL
Shipman
Department of Genetics, Harvard Medical School, Boston, MA, USA Wyss Institute for Biologically Inspired Engineering at Harvard University, Boston, MA, USA.
Susan M
SM
Byrne
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.
Neville E
NE
Sanjana
Department of Cell Biology, Harvard Medical School, Boston, MA, USA Division of Newborn Medicine, Boston Children's Hospital, Boston, MA, USA.
Jernej
J
Murn
Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA.
Yinqing
Y
Li
Department of Bioengineering, University of California, San Diego, CA, USA.
Shangzhong
S
Li
Friedrich Miescher Institute for Biomedical Research, Basel, Switzerland Swiss Institute of Bioinformatics, Basel, Switzerland University of Basel, Basel, Switzerland.
Michael
M
Stadler
Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA.
Ron
R
Weiss
Department of Genetics, Harvard Medical School, Boston, MA, USA Wyss Institute for Biologically Inspired Engineering at Harvard University, Boston, MA, USA gchurch@genetics.med.harvard.edu.
George M
GM
Church
D051792:Basic Helix-Loop-Helix Proteins / Q000378:metabolism*; D001921:Brain / Q000196:embryology / Q000378:metabolism; D002454:Cell Differentiation; D065150:Cellular Reprogramming; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000502:physiology*; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D055495:Neurogenesis; D015533:Transcriptional Activation
D051792:Basic Helix-Loop-Helix Proteins; D009419:Nerve Tissue Proteins; C105389:NEUROG1 protein, human; C491183:NEUROG2 protein, human
gene regulatory networks; microRNAs; neurogenesis; stem cell differentiation; transcriptomics