Neurogenin 2 enhances the generation of patient-specific induced neuronal cells

Brain Res. 2015 Jul 30;1615:51-60. doi: 10.1016/j.brainres.2015.04.027. Epub 2015 Apr 24.

Abstract

Direct lineage reprogramming of human fibroblasts into functional neurons holds great promise for biomedical applications such as regenerative medicine and cellbased disease modelling. However, clinical applications must consider how to increase neuronal conversion efficiency and at the same time reduce the number of required transcription factors. Here, we investigated whether Neurogenin 2 (Ngn2), which is a proneural gene that directs neuronal differentiation of progenitor cells during development, can enhance the generation of patient-specific induced neuronal cells. In this study, we transfected Ascl1, Sox2 and Ngn2 into human fibroblasts from the patients' scalp by lentivirus. Morphological analysis, immunocytochemistry, gene expression and electrophysiological analysis were performed to identify the similarity of induced neuronal cells (iNCs) to human neuronal cells. Ngn2 increase the conversion efficiency from 4% to 13.4%. iNCs express neuronal cell markers and resemble wild-type neurons in their morphology, gene expression profiles and exhibit functional membrane properties of mature neurons. Implanted iNCs can survive and integrate in mouse brains and, unlike iPS cell-derived neural cells, do not generate tumours.

Keywords: Induced neuron; Neurogenin 2; Reprogramming.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Basic Helix-Loop-Helix Transcription Factors / genetics
  • Basic Helix-Loop-Helix Transcription Factors / physiology*
  • Brain / cytology
  • Brain / physiology
  • Brain / surgery
  • Cells, Cultured
  • Cellular Reprogramming*
  • Gene Expression
  • Humans
  • Induced Pluripotent Stem Cells / cytology
  • Induced Pluripotent Stem Cells / metabolism
  • Induced Pluripotent Stem Cells / physiology*
  • Induced Pluripotent Stem Cells / transplantation
  • Mice
  • Mice, Inbred C57BL
  • Nerve Tissue Proteins / genetics
  • Nerve Tissue Proteins / physiology*
  • Neural Stem Cells / cytology
  • Neural Stem Cells / metabolism
  • Neural Stem Cells / physiology*
  • Neural Stem Cells / transplantation
  • SOXB1 Transcription Factors / genetics
  • SOXB1 Transcription Factors / physiology
  • Transplantation, Heterologous

Substances

  • ASCL1 protein, human
  • Basic Helix-Loop-Helix Transcription Factors
  • NEUROG2 protein, human
  • Nerve Tissue Proteins
  • SOX2 protein, human
  • SOXB1 Transcription Factors