Ciliary neurotrophic factor overexpression in neural progenitor cells (ST14A) increases proliferation, metabolic activity, and resistance to stress during differentiation

J Neurosci Res. 2003 Jan 15;71(2):228-36. doi: 10.1002/jnr.10477.

Abstract

Neurotrophic factors exert considerable neuroprotective and neurorestorative effects in neurodegenerative diseases. Because neuronal progenitor cells have, at least in part, the potency to restore degenerated neuronal networks, transgenic high-dosage expression of neurotrophins by these cells in neurotransplantation may be advantageous. In the present study, a retroviral vector containing the gene of rat ciliary neurotrophic factor (rCNTF) was permanently transfected into a striatal neuronal progenitor cell line. Qualitative and quantitative analyses demonstrated a sustained expression of the transgene; i.e., rCNTF was present at the mRNA level and protein level. Moreover, cocultivation in separate chambers of transgenic CNTF-ST14A cells and CNTF-dependent TF1 cells exerted typical biological effects, such as increased proliferation and differentiation of the TF1 cells, indicating the functional integrity of the secreted recombinant neurotrophin. The CNTF-ST14A cells displayed improved stress response compared with native ST14A cells under differentiation conditions, i.e., at the nonpermissive temperature of 39 degrees C and after staurosporine exposure, respectively. This effect coincided with a relatively reduced apoptosis rate and a raised metabolic activity of CNTF-ST14A cells at 39 degrees C. Neurotransplantation of CNTF-ST14A cells in the rat quinolinic acid model of Huntington's disease showed a significant and sustained decline in pathological apomorphine-induced rotations compared with parental ST14A cells. We conclude that sustained functional transgene CNTF production improves stress response as well as metabolic activity, making CNTF-ST14A cells a promising tool for neurotransplantation in the quinolinic acid model of Huntington's disease.

MeSH terms

  • Animals
  • Apomorphine / toxicity
  • Apoptosis / physiology
  • Blotting, Western
  • Cell Differentiation / physiology*
  • Cell Division / physiology
  • Cells, Cultured
  • Ciliary Neurotrophic Factor / metabolism*
  • Ciliary Neurotrophic Factor / physiology
  • Coculture Techniques
  • Corpus Striatum / injuries
  • Culture Media, Serum-Free / pharmacology
  • Disease Models, Animal
  • Enzyme Inhibitors / pharmacology
  • Fibroblasts / metabolism
  • Gene Expression
  • In Situ Nick-End Labeling
  • Quinolinic Acid / toxicity
  • RNA, Messenger / biosynthesis
  • Rats
  • Rats, Wistar
  • Reverse Transcriptase Polymerase Chain Reaction
  • Rotation
  • Staurosporine / pharmacology
  • Stem Cells / physiology*
  • Stress, Physiological*
  • Tetrazolium Salts
  • Thermosensing
  • Thymidine / biosynthesis
  • Time Factors
  • Transfection
  • Tritium

Substances

  • 2-(4-iodophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium
  • Ciliary Neurotrophic Factor
  • Culture Media, Serum-Free
  • Enzyme Inhibitors
  • RNA, Messenger
  • Tetrazolium Salts
  • Tritium
  • Quinolinic Acid
  • Staurosporine
  • Apomorphine
  • Thymidine