The zinc finger E-box-binding homeobox 1 (Zeb1) promotes the conversion of mouse fibroblasts into functional neurons

Long Yan; Yue Li; Zixiao Shi; Xiaoyin Lu; Jiao Ma; Baoyang Hu; Jianwei Jiao; Hongmei Wang

doi:10.1074/jbc.M116.771493

The zinc finger E-box-binding homeobox 1 (Zeb1) promotes the conversion of mouse fibroblasts into functional neurons

¹ Institute of Zoology, Chinese Academy of Sciences, China;
² Peking University Third Hospital, China

↵* Corresponding author; email: wanghm{at}ioz.ac.cn

Author contributions: Author contributions: H.W. conceived the study, designed the experiments, and supervised the research. J.J. conceived the study and critically revised the manuscript. L.Y., Y.L., Z.S. and X.L. conducted the experiments and analyzed the data. L.Y. and Y.L. collected and analyzed the data, organized figures and drafted the manuscript. Z.S. and X.L. contributed to the adenoviral vector construction and the figure organization. J.M. contributed to the immunofluorescent experiments. B.H. conceived the study and commented on the manuscript. All authors contributed to the figure and manuscript preparation and approved the version that was submitted.

Abstract

The zinc finger E-box binding transcription factor Zeb1 plays a pivotal role in the epithelial-mesenchymal transition (EMT). Numerous studies have focused on the molecular mechanisms by which Zeb1 contributes to this process. However, the functions of Zeb1 beyond EMT remain largely elusive. Using a transdifferentiation system to convert mouse embryonic fibroblasts (MEFs) into functional neurons via the neuronal transcription factors Achaete-scute family bHLH transcription factor1 (Ascl1), POU class 3 homeobox 2 (POU3F2/Brn2) and Neurogenin 2 (Neurog2, Ngn2) (ABN), we found that Zeb1 was up-regulated during the early stages of transdifferentiation. Knocking down Zeb1 dramatically attenuated the transdifferentiation efficiency, whereas Zeb1 overexpression obviously increased the efficiency of transdifferentiation from MEFs to neurons. Interestingly, Zeb1 improved the transdifferentiation efficiency induced by even a single transcription factor (e.g., Asc1 or Ngn2). Zeb1 also rapidly promoted the maturation of induced neuron (iN) cells to functional neurons and improved the formation of neuronal patterns and electrophysiological characteristics. iN cells could form functional synapse in vivo after transplantation. Genome-wide RNA arrays showed that Zeb1 overexpression up-regulated the expression of neuron-specific genes and down-regulated the expression of epithelial-specific genes during conversion. Taken together, our results reveal a new role for Zeb1 in the transdifferentiation of MEFs into neurons.