Ciliary neurotrophic factor overexpression in neural progenitor cells (ST14A) increases proliferation, metabolic activity, and resistance to stress during differentiation.
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.
Weinelt
Sabine
S
Neurobiological Laboratory, Department of Neurology, University of Rostock, Rostock, Germany.
Peters
Sabine
S
Bauer
Peter
P
Mix
Eilhard
E
Haas
Stefan J P
SJ
Dittmann
Aline
A
Petrov
Stanislov
S
Wree
Andreas
A
Cattaneo
Elena
E
Knoblich
Rupert
R
Strauss
Ulf
U
Rolfs
Arndt
A
D000818:Animals; D001058:Apomorphine / Q000633:toxicity; D017209:Apoptosis / Q000502:physiology; D015153:Blotting, Western; D002454:Cell Differentiation / Q000502:physiology*; D002455:Cell Division / Q000502:physiology; D002478:Cells, Cultured; D020934:Ciliary Neurotrophic Factor / Q000378:metabolism* / Q000502:physiology; D018920:Coculture Techniques; D003342:Corpus Striatum / Q000293:injuries; D016895:Culture Media, Serum-Free / Q000494:pharmacology; D004195:Disease Models, Animal; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005347:Fibroblasts / Q000378:metabolism; D015870:Gene Expression; D020287:In Situ Nick-End Labeling; D017378:Quinolinic Acid / Q000633:toxicity; D012333:RNA, Messenger / Q000096:biosynthesis; D051381:Rats; D017208:Rats, Wistar; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012399:Rotation; D019311:Staurosporine / Q000494:pharmacology; D013234:Stem Cells / Q000502:physiology*; D013312:Stress, Physiological; D013778:Tetrazolium Salts; D013697:Thermosensing; D013936:Thymidine / Q000096:biosynthesis; D013997:Time Factors; D014162:Transfection; D014316:Tritium
C110656:2-(4-iodophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium; D020934:Ciliary Neurotrophic Factor; D016895:Culture Media, Serum-Free; D004791:Enzyme Inhibitors; D012333:RNA, Messenger; D013778:Tetrazolium Salts; D014316:Tritium; D017378:Quinolinic Acid; D019311:Staurosporine; D001058:Apomorphine; D013936:Thymidine