Rescue of lesioned adult rat spinal motoneurons by adenoviral gene transfer of glial cell line-derived neurotrophic factor

Abstract

Glial cell line-derived neurotrophic factor (GDNF) has been shown to protect cranial and spinal motoneurons, that suggests potential uses of GDNF in the treatment of spinal cord injury and motor neuron diseases. We examined neuroprotective effect of human GDNF encoded by an adenovirus vector (AxCAhGDNF) on the death of lesioned adult rat spinal motoneurons. The seventh cervical segment (C7) ventral and dorsal roots and dorsal root ganglia of adult Fisher 344 rats were avulsed, and AxCAhGDNF, AxCALacZ (adenovirus encoding beta-galactosidase gene) or PBS was inoculated in C7 vertebral foramen. One week after the avulsion and treatment with AxCALacZ, the animals showed expression of beta-galactosidase activity in lesioned spinal motoneurons. Animals avulsed and treated with AxCAhGDNF showed intense immunolabeling for GDNF in lesioned spinal motoneurons and expression of virus-induced human GDNF mRNA transcripts in the lesioned spinal cord tissue. Nissl-stained cell counts revealed that the treatment with AxCAhGDNF significantly prevented the loss of lesioned ventral horn motoneurons 2 to 8 weeks after avulsion, as compared to AxCALacZ or PBS treatment. Furthermore, the AxCAhGDNF treatment ameliorated choline acetyltransferase immunoreactivity in the lesioned motoneurons after avulsion. These results indicate that the adenovirus-mediated gene transfer of GDNF may prevent the degeneration of motoneurons in adult humans with spinal cord injury and motor neuron diseases.

Intrathecal transplantation of human neural stem cells overexpressing VEGF provide behavioral improvement, disease onset delay and survival extension in transgenic ALS mice

“…38,[41][42][43] Of these neurotrophic factors, VEGF is well known to have an important function in the pathogenesis of ALS. [31][32][33][34]37,38 The genetically modified human NSCs we established, F3.VEGF cells, were confirmed to release four times of VEGF165, the most abundant and biologically active isoform in vivo, compared with the parental F3 cells as we have reported previously in animal models of stroke.…”

Adenoviral GDNF gene transfer enhances neurofunctional recovery after recurrent laryngeal nerve injury

“…[28][29][30]46 The present study analyzed neurological function using electrophysiological examination in vivo and demonstrated neurofunctional recovery. This report carries a higher clinical impact than our previous reports.…”

Adenovirus-mediated overexpression of a gene prevents hearing loss and progressive inner hair cell loss after transient cochlear ischemia in gerbils

“…Briefly, human GDNF cDNA derived from cultured human fetal astrocytes was subcloned into a cassette cosmid pAxCAwt carrying an adenovirus type-5 genome lacking the E3, E1A, and E1B regions to prevent virus replication. 35,36 Cosmid pAxCAwt contains a Swa I cloning site flanked by a cytomegalovirus-enhancerchicken b-actin hybrid (CAG) promoter at the 5 0 end, and a rabbit globin poly(A) sequence at the 3 0 end. The cosmid was cotransfected into 293 cells with the appropriately cleaved adenovirus genome lacking the E3 region.…”