Details

Autor(en) / Beteiligte

• Busskamp, Volker
• Lewis, Nathan E
• Guye, Patrick
• Ng, Alex HM
• Shipman, Seth L
• Byrne, Susan M
• Sanjana, Neville E
• Murn, Jernej
• Li, Yinqing
• Li, Shangzhong
• Stadler, Michael
• Weiss, Ron
• Church, George M

Titel

Rapid neurogenesis through transcriptional activation in human stem cells

Ist Teil von

• Molecular systems biology, 2014-11, Vol.10 (11), p.760-n/a

Ort / Verlag

England: EMBO Press

Erscheinungsjahr

2014

Quelle

Wiley-Blackwell Journals

Beschreibungen/Notizen

• 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. 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.