HSV amplicon-mediated neurotrophin-3 expression protects murine spiral ganglion neurons from cisplatin-induced damage.

Abstract

Ototoxicity is a major dose-limiting side effect of cisplatin (DDP) administration due to its propensity to induce destruction of hair cells and neurons in the auditory system. Previous studies demonstrated that TrkC-expressing spiral ganglion neurons (SGN) are protected from the cytotoxic effects of DDP by localized delivery of the trophic factor neurotrophin-3 (NT-3). Successful in vivo implementation of such a therapy requires the development of an efficient gene delivery vehicle for expression of NT-3 within the cochlea. To this end, we constructed a herpes simplex virus (HSV) amplicon vector that expressed a c-Myc-tagged NT-3 chimera (HSVnt-3myc). Helper virus-free vector stocks were initially evaluated in vitro for their capacity to direct expression of NT-3 mRNA and protein. Transduction of cultured murine cochlear explants with HSVnt-3myc resulted in production of NT-3 mRNA and protein up to 3 ng/ml as measured over a 48-h period in culture supernatants. To determine whether NT-3 overexpression could abrogate DDP toxicity, cochlear explants were transduced with HSVnt-3myc or a murine intestinal alkaline phosphatase-expressing control vector, HSVmiap, and then exposed to cisplatin. HSVnt-3myc-transduced cochlear explants harbored significantly greater numbers of surviving SGNs than those infected with control virus. These data demonstrate that amplicon-mediated NT-3 transduction can attenuate the ototoxic action of DDP on organotypic culture. The potency of NT-3 in protecting spiral ganglion neurons from degeneration suggests that in vivo neurotrophin-based gene therapy may be useful for the prevention and/or treatment of hearing disorders.

• Authors: X Chen, R D Frisina, W J Bowers, D R Frisina, H J Federoff
• Journal: Molecular therapy : the journal of the American Society of Gene Therapy (Mol Ther) Vol. 3 Issue 6 Pg. 958-63 (Jun 2001) ISSN: 1525-0016 [Print] United States
• PMID: 11407910 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, P.H.S.)

Chemical References

• DNA Primers
• Neurotrophin 3
• Recombinant Fusion Proteins
• Cisplatin

Topics

• Animals
• Blotting, Western
• Cell Death (drug effects)
• Cisplatin (toxicity)
• Cochlea (metabolism)
• Cytomegalovirus (genetics)
• DNA Primers (chemistry)
• Enzyme-Linked Immunosorbent Assay
• Genes, myc (genetics, physiology)
• Hearing Disorders (chemically induced, prevention & control)
• Immunoenzyme Techniques
• Neurites (chemistry)
• Neurons (drug effects, pathology)
• Neurotrophin 3 (genetics, metabolism)
• Recombinant Fusion Proteins (chemistry, genetics, metabolism)
• Reverse Transcriptase Polymerase Chain Reaction
• Simplexvirus (genetics)
• Spiral Ganglion (drug effects, pathology)